ORIGINAL ARTICLE
Association Between Cyclooxygenase-2 Gene Polymorphisms and Head and Neck Squamous Cell Carcinoma Risk Yuming Niu, MD,* Hua Yuan, MD,Þ Ming Shen, MD,Þ Huizhang Li, MD,þ Yuanyuan Hu, MD,* and Ning Chen, PhDÞ Abstract: Cyclooxygenase-2 (COX-2) is an inducible enzyme that catalyzes prostaglandins through inflammatory response, which may be involved in autoimmune diseases and cancer pathogenesis. Two potentially functional genetic single-nucleotide polymorphisms (COX-2 -1195G9A and 8473T9C) were supposed to contribute to head and neck squamous cell carcinoma (HNSCC) susceptibility. The aim of this study was to determine the association of these polymorphisms with HNSCC susceptibility in a Chinese Han population. In this study, 2 SNPs were genotyped by TaqMan methods in a patient-control study including 260 patients with HNSCC and 1047 cancer-free controls in a Chinese Han population. We found significant difference in the frequency of alcohol consumption between the patients with HNSCC and controls (P G 0.001), but the genotype frequencies of the 2 polymorphisms were not significantly different between the patients and controls. Further stratified analysis indicated that none of the genotypes were associated with increased risk for HNSCC. This research indicated that the COX-2 -1195G9A and 8473T9C polymorphisms may not be involved in the development of HNSCC in the Chinese Han population. However, further perspective studies are warranted to test these findings and further investigate the potential interactions involving the COX-2 polymorphism and HNSCC.
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From the *Department of Stomatology, Taihe Hospital, Hubei University of Medicine, Shiyan; †Institute of Dental Research, and ‡Department of Epidemiology and Biostatistics, School of Public Health, Nanjing Medical University, Nanjing, People’s Republic of China. Received November 16, 2012. Accepted for publication August 26, 2013. Address correspondence and reprint requests to Ning Chen, PhD, Institute of Dental Research, Nanjing Medical University, 140 Hanzhong Rd, Nanjing, 210029, People’s Republic of China; E-mail: [email protected] Supported by grants from the Foundation of Taihe Hospital of Hubei University of Medicine (2011QD05, 2010D22) and the Medical Development Foundation of Health Department of Jiangsu Province (H200811). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the article. The authors report no conflicts of interest. Copyright * 2014 by Mutaz B. Habal, MD ISSN: 1049-2275 DOI: 10.1097/SCS.0000000000000372
The Journal of Craniofacial Surgery
Key Words: COX-2, polymorphism, head and neck squamous cell carcinoma, susceptibility (J Craniofac Surg 2014;25: 333Y337)
H
ead and neck squamous cell carcinoma (HNSCC) encompasses epithelial malignancies that arise in the paranasal sinuses, nasal cavity, oral cavity, pharynx, and larynx.1 It is the sixth most common malignancy worldwide, which represents approximately 6% of all cases and accounts for approximately 650,000 new cancer cases and 350,000 cancer deaths every year.2,3 Tobacco and alcohol consumption and other environmental sources have been implicated as the important causes of HNSCC development.4,5 Inflammation is an important process in the development of cancer. Prostaglandins are believed to play a key role in inflammation and contribute to cancer development and progression through cellular proliferation and angiogenesis.6,7 Cyclooxygenase (COX, also known as prostaglandin endoperoxide synthases) is a rate-limiting enzyme in conversion of arachidonic acid into prostaglandins. The COX family is composed of 2 isozymes: COX-1 and COX-2.8 Cyclooxygenase-1 is always constitutively expressed and involved in normal physiological functions, whereas COX-2 is an inducible enzyme that catalyzes prostaglandins via inflammatory responses, growth factors, cytokines, and various carcinogens.9,10 Overexpression of COX-2 may be associated with cellular proliferation, inhibition of apoptosis, enhanced invasiveness of cancer cells, and a promotion of angiogenesis11,12 and has been observed in many cancers, especially in the upper aerodigestive tract cancers, such as oral cancer, gastric cancer, and esophageal cancer.13Y15 Some COX-2 inhibitors (eg, nonsteroidal anti-inflammatory drugs, nonsteroidal anti-inflammatory drugs), such as aspirin, indomethacin, and NS398, could inhibit the proliferation of cancer cells independently through the reduction of COX-2 protein expression and suppress the invasiveness of squamous cell carcinoma cell lines via downregulation of matrix metalloproteinase-2 production and activation.16,17 Environmental and genetic risk factors are known to be responsible for HNSCC and act either independently or in combination.3 Interindividual variations in the COX-2 gene may influence the expression level and activity of the protein and subsequently change the susceptibility to developing HNSCC. Two potentially functional single-nucleotide polymorphisms (SNPs), -1195G9A (rs689466) and 8473T9C (rs5275), are located in the promoter region and 3’ untranslated region (UTR) of the COX-2 gene, respectively. These SNPs could affect gene transcription and messenger RNA (mRNA) stability. Lots of molecular epidemiological studies have been conducted to examine the association between these polymorphisms and the susceptibility to HNSCC in diverse populations, but the results remain controversial.18Y23 In this study, we conducted a patientcontrol study to investigate the frequencies of SNPs of COX-2
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between patients with HNSCC and controls in the Chinese Han population and evaluate their contribution to the risk for developing HNSCC.
MATERIALS AND METHODS Study Participants The study was approved by the institutional review board of the Nanjing Medical University, Nanjing, China. All of the participants were genetically unrelated ethnic Han Chinese in Jiangsu province. No restrictions in age, sex, and histology were set, and exclusion criteria were as follows: previous cancer, metastasized cancer, and previous radiotherapy or chemotherapy. A total of 260 patients with newly diagnosed and histologically confirmed squamous cell carcinoma (SCC) of the oral (including the oral cavity and oropharynx) and larynx were recruited from 2008 to 2011 at the Jiangsu Provincial Stomatological Hospital, the Cancer Hospital of Jiangsu Province, and the First Affiliated Hospital of Nanjing Medical University, Nanjing, China. In addition, 1047 cancer-free controls were randomly selected from a pool of 30,000 individuals who participated in a communitybased screening program for noninfectious diseases conducted in Jiangsu province during the same period when the patients were recruited. The control participants had no history of cancer and were frequency matched to the patients by age (T5 y), sex, and residential area (urban or rural). After written informed consent had been obtained, face-to-face interviews were conducted to obtain demographic data (eg, age and sex) and exposure information (eg, smoking status) by trained interviewers. Approximately 5 mL of venous blood sample was collected from each participant. Individuals who smoked 1 cigarette per day for 1 year were defined as ever smokers; otherwise, they were considered as never smokers. In addition, those who consumed 3 or more alcoholic drinks a week for more than 6 months were considered alcohol drinkers.24
Genotyping Genomic DNA was extracted from a leukocyte pellet by proteinase K digestion and followed by phenol-chloroform extraction and ethanol precipitation. All SNPs were analyzed by the conventional TaqMan minor groove binder method. The primers and probes for -1195G9A are as follows: primer: sense, 5’-GCACTACCCATGATAGATGTTAAACAA-3’, antisense, 5’-GGAATTAATTAGATGGAAGGGAGATTT-3’; probe: allele A, FAM-ATGAAATTCCAACTGTC-MGB, allele G, HEX-TGAAAT TCCAGCTGTC-MGB. The primers and probes for 8473T9C are as follows: primer: sense, 5’-TTCCAATGCATCTTCCATGATG-3’, antisense, 5’-ATGCACTGATACCTGTTTTTGTTTG-3’; probe: allele T, FAM-AGTACTTTTGGTTATTTT-MGB, allele C, HEX-AGTA CTTTTGGTCATTTT-MGB. Genotyping was performed without
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TABLE 1. Characteristic Information Between the Patients With HNSCC and Controls Variables Age Mean (SD) e60 960 Sex Male Female Smoking status Never Ever Drinking status Never Ever Tumor location Oral cancer Laryngeal cancer
Patients, n (%)
Controls, n (%)
P
60.41 (8.23) 146 (56.16) 114 (43.84)
59.57 (11.13) 549 (52.44) 498 (47.56)
0.282
180 (69.23) 80 (30.77)
771 (73.64) 276 (26.36)
0.153
134 (51.54) 126 (48.46)
503 (48.04) 544 (51.96)
0.313
135 (51.92) 125 (48.08)
681 (68.04) 366 (34.96)
G0.001
169 (65.00) 91 (35.00)
knowing the patient’s case or control’s status. Ten percent of samples were randomly selected to repeat to validate genotype identification, and the results were totally concordant.
Statistical Analysis The differences in the distributions of demographic characteristics, selected variables, and genotypes of the COX-2 variants between the patients and controls were evaluated with Student’s t-test or Chi-squared tests. The Hardy-Weinberg equilibrium of the genotype distribution of the controls was tested by a goodness-of-fit Chi-squared test. Linkage disequilibrium was estimated with an online software platform.25 The associations between COX-2 SNPs and HNSCC risk were estimated by computing the odds ratios (ORs) and their 95% confidence intervals (CIs) from both univariate and multivariate logistic regression analyses without and with the adjustments for age, sex, smoking status, and drinking status. All statistical analyses were performed with SAS 9.1.3 software (SAS Institute, Cary, NC). Statistical significance was set at P G 0.05, and all statistical tests were 2-sided.
RESULTS The demographic characteristics and clinical information for the 260 patients with HNSCC (mean [SD] age, 60.41 [8.23]) and 1047 controls (mean [SD] age, 59.57 [11.13]) enrolled in our study are shown in Table 1. Approximately 48.46% of the patients with
TABLE 2. Odds Ratios and Confidence Intervals of Associations Between COX-2 SNPs and HNSCC (Oral SCC and Laryngeal SCC)
Genotypes
Controls, n (%)
All Patients, n (%)
-1195G9A GG GA AA GA+AA 8473T9C TT TC CC TC+CC
1035 222 (21.45) 542 (52.37) 271 (26.18) 813 (78.55) 1032 691 (66.96) 316 (30.62) 25 (2.42) 341 (33.04)
259 (23.55) (48.65) (27.80) (76.45) 258 177 (68.60) 72 (27.91) 9 (3.49) 81 (31.40) 61 126 72 198
Adjusted OR (95% CI)*
1.00 0.85 (0.60Y1.21) 1.01 (0.69Y1.50) 0.91 (0.65Y1.26) 1.00 0.90 (0.66Y1.22) 1.48 (0.68Y3.25) 0.94 (0.70Y1.26)
Oral SCC, n (%) 169 (26.04) (47.34) (26.63) (73.96) 168 118 (70.24) 45 (26.79) 5 (2.98) 50 (29.76)
44 80 45 125
Adjusted OR (95% CI)*
1.00 0.74 (0.49Y1.11) 0.87 (0.55Y1.39) 0.78 (0.53Y1,14) 1.00 0.86 (0.58Y1.26) 1.03 (0.36Y2.97) 0.87 (0.60Y1.27)
Laryngeal SCC, n (%) 90 (18.89) (51.11) (30.00) (81.11) 90 59 (65.56) 27 (30.00) 4 (4.44) 31 (34.44) 17 46 27 73
Adjusted OR (95% CI)*
1.00 1.16 (0.65Y2.09) 1.43 (0.75Y2.75) 1.23 (0.71Y2.15) 1.00 1.02 (0.63Y1.64) 1.62 (0.54Y4.88) 1.07 (0.67Y1.69)
*Adjusted by age, sex, smoking status, and drinking status.
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Cyclooxygenase-2 Polymorphism and HNSCC
TABLE 3. Stratified Analysis on the Association Between COX-2 SNPs and HNSCC Risk -1195G9A Patient (n = 259) Characteristics Age, y e60 960 Sex Male Female Smoking status Never Ever Drinking status Never Ever
8473T9C
Control (n = 1035)
Patient (n = 258)
Control (n = 1032)
GG
GA+AA
GG
GA+AA
OR (95% CI)*
TT
TC+CC
TT
TC+CC
OR (95% CI)*
36 (24.83) 25 (21.93)
109 (75.17) 89 (78.07)
114 (20.99) 108 (21.95)
429 (79.01) 384 (78.05)
0.85 (0.55,1.32) 1.01 (0.61, 1.67)
99 (68.28) 78 (69.03)
46 (31.72) 35 (30.97)
375 (69.19) 316 (64.49)
167 (30.81) 174 (35.51)
1.08 (0.72, 1.61) 0.80 (0.51, 1.25)
40 (22.35) 21 (26.25)
139 (77.65) 59 (73.75)
161 (21.18) 61 (22.18)
599 (78.82) 214 (77.82)
0.99 (0.66, 1.49) 0.87 (0.48, 1.58)
124 (68.89) 53 (67.95)
56 (31.11) 25 (32.05)
509 (67.06) 182 (66.67)
250 (32.94) 91 (33.33)
0.96 (0.67, 1.37) 1.10 (0.63, 1.92)
21 (22.58) 40 (24.10)
72 (77.42) 126 (75.90)
104 (20.88) 118 (21.97)
394 (79.12) 419 (78.03)
1.05 (0.60, 1.81) 0.94 (0.62, 1.44)
63 (69.23) 114 (68.26)
28 (30.77) 53 (31.74)
333 (67.00) 358 (66.92)
164 (33.00) 177 (33.08)
0.95 (0.57, 1.56) 1.04 (0.71, 1.53)
21 (21.65) 40 (24.69)
76 (78.35) 122 (75.31)
111 (22.98) 111 (20.11)
372 (77.02) 441 (79.89)
1.15 (0.67, 1.98) 0.87 (0.57, 1.34)
66 (69.47) 111 (68.10)
29 (30.53) 52 (31.90)
319 (66.18) 372 (67.64)
163 (33.82) 178 (32.36)
0.90 (0.55, 1.48) 1.06 (0.72, 1.57)
*Adjusted by age, sex, smoking status, and drinking status.
HNSCC were smokers, which is not significantly different from that of the controls (51.96%) (P 9 0.05); otherwise, the HNSCC group had significantly more drinkers (48.08% versus 34.96%, P G 0.001) than those in the control group. The genotype distributions of the 2 SNPs in the patients and controls are shown in Table 2. The observed genotype frequencies for the 2 polymorphisms in the controls were all in Hardy-Weinberg equilibrium (P = 0.110 and 0.112 for -1195G9A and 8473T9C, respectively), which indicates good homogeneity within the study participants. One patient (0.38%) and 12 controls (1.15%) for -1195G9A as well as 2 patients (0.77%) and 12 controls (1.24%) for 8473T9C were not able to be genotyped owing to poor DNA quantity and/or quality. For -1195G9A, the genotype frequencies of GG, GA, and AA in the controls were 21.45%, 52.37%, and 26.18%, respectively, whereas those of the patients with HNSCC were 23.55%, 48.65%, and 27.80%, respectively (P = 0.555). Similarly, no statistically significant differences among the genotype frequencies of 8473T9C were observed (P = 0.478). In the multivariate logistic regression analysis, neither the heterozygote (for -1195G9A GA genotype and 8473T9C TC genotype) nor the mutated homozygote (for -1195G9A AA genotype and 8473T9C CC genotype) of the SNPs was associated with the developing risk for HNSCC, compared with their corresponding wild homozygote (for -1195G9A GG genotype and 8473T9C TT genotype). When the heterozygote and mutated homozygote were combined as the dominant model (for -1195G9A GA/AA genotype and 8473T9C TC/CC genotype), nonsignificant associations were observed between the variant genotypes and HNSCC risk (Table 2). In addition, further stratified analysis of -1195G9A and 8473T9C suggested no significant association with HNSCC after adjusting for age, sex, smoking status, and drinking status (Table 3). The estimation of linkage disequilibrium showed that -1195G9A and 8473T9C were not in strong linkage disequilibrium with the standardized D’= 0.978 and r2 = 0.189. We thus considered the 2 SNPs together but found no significant association for HNSCC with the combined COX-2 genotypes (Table 4).
DISCUSSION Epidemiological studies have demonstrated that the development of HNSCC may be largely caused by alcohol and tobacco use.5,26 An increase of approximately 6-fold in HNSCC risk has been
observed among smokers compared with nonsmokers.3,27 Furthermore, the risk for developing HNSCC apparently increases if smokers are also heavy drinkers,4,28 although our research showed that characteristic information does not indicate significant associations between smoking and HNSCC. On the other hand, an almost 2-fold increased risk for HNSCC has been observed in drinkers. As we know, acetaldehyde can generate from ethanol through oxidation by alcohol dehydrogenase, which is a strong risk factor for the development of HNSCC.29,30 Alcohol dehydrogenase can induce gene mutations, especially inactive variant aldehyde dehydrogenase-2 genotype to increase the accumulation of acetaldehyde after alcohol consumption.31,32 Furthermore, some new evidences also indicate that the human papilloma virus infection, especially that by human papilloma virus type 16, is a risk factor for HNSCC.33,34 The COX-2 gene that encodes the COX-2 enzyme has been mapped to chromosome 1q25.2Y25.3. Cyclooxygenase-2 is a key enzyme produced during the conversion of free arachidonic acid into prostaglandins, which is relevant to cancer development and progression.35 Furthermore, the increased expression of COX-2 could be involved in tumorigenesis by enhancing tumor cell proliferation, angiogenesis, inhibition of apoptosis, immune escape, and increased metastasis.36 Meanwhile, the expression and stability of COX-2 mRNA will be regulated by various elements of the 5’ promoter region and 3’UTR of the transcript.37 The SNP -1195G9A, which is located in the promoter region, may create a c-MYB binding site with strikingly high promoter activity.38 Another SNP located in the 3’UTR, 8473T9C, will alter mRNA stability as well as translation efficiency and subsequently influences cancers susceptibility.39,40
TABLE 4. Locus-Locus Interaction Between -1195G9A and 8473T9C on HNSCC Risk Genotypes
Patient -1195G9A 8473T9C (n = 257)
GG GG GA+AA GA+AA
Control (n = 1029)
OR (95 CI%)
OR (95 CI%)*
TT 60 (23.35) 220 (21.38) 1.00 1.00 TC+CC 1 (0.39) 2 (0.19) 1.84 (0.16Y20.5) 1.67 (0.14Y19.45) TT 116 (45.14) 468 (45.48) 0.91 (0.64Y1.29) 0.93 (0.65Y1.32) TC+CC 80 (31.13) 339 (32.94) 0.87 (.060Y1.26) 0.89 (.061Y1.30)
*Adjusted by age, sex, smoking status, and drinking status.
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Chiang et al21 recently conducted a patient-control study of 377 patients with oral SCC and 442 controls in a China Taiwan population and found that COX-2 -1195AA genotypes are associated with a significant 1.55-fold (95% confidence interval [CI], 1.04Y2.31) increase in risk for developing oral cancer compared with their wildtype genotypes. However, patient-control studies in a white and Indian populations showed that the -1195G9A polymorphism is not significantly associated with HNSCC risk.19,37 Moreover, using dual-luciferase reporter analysis with plasmids containing the 1195G9A SNP to analyze transcription activity, Shi et al41 and Sakaki et al42 reported that the promoter region containing -1195G shows significantly higher transcription activity than that containing 1195A. However, a study performed by Zhang et al38 suggested opposing findings. In molecular epidemiological studies on the association between COX-2 8473T9C polymorphism and cancer susceptibility, results conflicted among different types of cancer and no significant association with HNSCC was found.23,37 This patient-control study comprised 260 patients with HNSCC and 1047 healthy controls in the Chinese Han population. Similar frequencies of the alleles and genotypes were found, and none of the genotypes were associated with increased risks for HNSCC, which suggests that no genotype contributes to the development of HNSCC in the Chinese Han population. To improve the power of genetic analysis and understand the results of risk assessment, we stratified all patients with HNSCC into 2 subgroups (oral cancer and laryngeal cancer). Further stratified analyses (age, sex, smoking status, drinking status, and haplotype frequency assessments) were also performed. The genotype frequency was similarly distributed between each of the subgroups and the controls. None of the risk genotypes were found to be in association with any subgroup of HNSCC. Several limitations in our study must be addressed. First, other SNP loci, such as -765 G9C in the promoter region of the COX-2 gene, have been related to cancer risk. Unfortunately, because of technical reasons, we were unable to detect the genotype of the -765 G9C polymorphism in the patient and control groups successfully. The association between -765 G9C polymorphism and HNSCC risk as well as the combined effect of the haplotype of -765 G9C and -1195G9A polymorphism in the promoter region were not observed in this research. Second, the sample size of the patients with HNSCC was not sufficiently large and information from some patients was not collected completely during the early stage of this research. The association between the -1195G9A polymorphism and cancer subtypes or tumor stages was not explored. This limitation could reduce the validity of the research findings because some potentially hidden risk factors may not have been fully exposed. Third, inherent biases may have led to spurious findings because the patients were recruited from the hospital and the controls were selected from a similar population. Despite these limitations, the potential bias was considered minimal and had no influence on the results because the controls were matched to the patients with HNSCC on the basis of age, sex, as well as residential area and drinking and smoking statuses were adjusted during the statistical analyses. In conclusion, the current study did not find any association between the 2 functional genetic variants of the COX-2 gene and HNSCC risk in the Chinese Han population. Well-designed studies are further warranted to validate these findings and investigate the potential gene-gene and gene-environment interactions involved in COX-2 polymorphisms and HNSCC.
REFERENCES 1. Argiris A, Eng C. Epidemiology, staging, and screening of head and neck cancer. Cancer Treat Res 2003;114:15Y60 2. Parkin DM, Bray F, Ferlay J, et al. Global cancer statistics, 2002. CA Cancer J Clin 2005;55:74Y108
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3. Argiris A, Karamouzis MV, Raben D, et al. Head and neck cancer. Lancet 2008;371:1695Y1709 4. Blot WJ, McLaughlin JK, Winn DM, et al. Smoking and drinking in relation to oral and pharyngeal cancer. Cancer Res 1988;48:3282Y3287 5. Vineis P, Alavanja M, Buffler P, et al. Tobacco and cancer: recent epidemiological evidence. J Natl Cancer Inst 2004;96:99Y106 6. Cao Y, Prescott SM. Many actions of cyclooxygenase-2 in cellular dynamics and in cancer. J Cell Physiol 2002;190:279Y286 7. Romano M, Claria J. Cyclooxygenase-2 and 5-lipoxygenase converging functions on cell proliferation and tumor angiogenesis: implications for cancer therapy. FASEB J 2003;17:1986Y1995 8. Simmons DL, Botting RM, Hla T. Cyclooxygenase isozymes: the biology of prostaglandin synthesis and inhibition. Pharmacol Rev 2004;56:387Y437 9. Vane SJ. Differential inhibition of cyclooxygenase isoforms: an explanation of the action of NSAIDs. J Clin Rheumatol 1998;4:s3Ys10 10. Herschman HR. Prostaglandin synthase 2. Biochim Biophys Acta 1996;1299:125Y140 11. Souza RF, Shewmake K, Beer DG, et al. Selective inhibition of cyclooxygenase-2 suppresses growth and induces apoptosis in human esophageal adenocarcinoma cells. Cancer Res 2000;60:5767Y5772 12. Ranger GS, Thomas V, Jewell A, et al. Elevated cyclooxygenase-2 expression correlates with distant metastases in breast cancer. Anticancer Res 2004;24:2349Y2351 13. Bayazit YA, Buyukberber S, Sari I, et al. Cyclo-oxygenase 2 expression in laryngeal squamous cell carcinoma and its clinical correlates. ORL J Otorhinolaryngol Relat Spec 2004;66:65Y69 14. Segawa E, Sakurai K, Kishimoto H, et al. Expression of cyclooxygenase-2 and DNA topoisomerase II alpha in precancerous and cancerous lesions of the oral mucosa. Oral Oncol 2008;44:664Y671 15. Liu X, Li P, Zhang ST, et al. COX-2 mRNA expression in esophageal squamous cell carcinoma (ESCC) and effect by NSAID. Dis Esophagus 2008;21:9Y14 16. Gao J, Niwa K, Sun W, et al. Non-steroidal anti-inflammatory drugs inhibit cellular proliferation and upregulate cyclooxygenase-2 protein expression in endometrial cancer cells. Cancer Sci 2004;95:901Y907 17. Kurihara Y, Hatori M, Ando Y, et al. Inhibition of cyclooxygenase-2 suppresses the invasiveness of oral squamous cell carcinoma cell lines via down-regulation of matrix metalloproteinase-2 production and activation. Clin Exp Metastasis 2009;26:425Y432 18. Jeong SW, Tae K, Lee SH, et al. Cox-2 and IL-10 polymorphisms and association with squamous cell carcinoma of the head and neck in a Korean sample. J Korean Med Sci 2010;25:1024Y1028 19. Peters WH, Lacko M, Te Morsche RH, et al. COX-2 polymorphisms and the risk for head and neck cancer in white patients. Head Neck 2009;31:938Y943 20. Pu X, Lippman SM, Yang H, et al. Cyclooxygenase-2 gene polymorphisms reduce the risk of oral premalignant lesions. Cancer 2009;115:1498Y1506 21. Chiang SL, Chen PH, Lee CH, et al. Up-regulation of inflammatory signalings by areca nut extract and role of cyclooxygenase-2 -1195G9A polymorphism reveal risk of oral cancer. Cancer Res 2008;68: 8489Y8498 22. Lin YC, Huang HI, Wang LH, et al. Polymorphisms of COX-2 -765G9C and p53 codon 72 and risks of oral squamous cell carcinoma in a Taiwan population. Oral Oncol 2008;44:798Y804 23. Campa D, Hashibe M, Zaridze D, et al. Association of common polymorphisms in inflammatory genes with risk of developing cancers of the upper aerodigestive tract. Cancer Causes Control 2007;18:449Y455 24. Chu H, Cao W, Chen W, et al. Potentially functional polymorphisms in IL-23R and risk of esophageal cancer in a Chinese population. Int J Cancer 2011;130:1093Y1097 25. Shi YY, He L. SHEsis, a powerful software platform for analyses of linkage disequilibrium, haplotype construction, and genetic association at polymorphism loci. Cell Res 2005;15:97Y98 26. Secretan B, Straif K, Baan R, et al. A review of human carcinogensVPart E: tobacco, areca nut, alcohol, coal smoke, and salted fish. Lancet Oncol 2009;10:1033Y1034
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27. Kamangar F, Dores GM, Anderson WF. Patterns of cancer incidence, mortality, and prevalence across five continents: defining priorities to reduce cancer disparities in different geographic regions of the world. J Clin Oncol 2006;24:2137Y2150 28. Smith EM, Rubenstein LM, Haugen TH, et al. Tobacco and alcohol use increases the risk of both HPV-associated and HPV-independent head and neck cancers. Cancer Causes Control 2010;21:1369Y1378 29. Guha N, Boffetta P, Wunsch Filho V, et al. Oral health and risk of squamous cell carcinoma of the head and neck and esophagus: results of two multicentric case-control studies. Am J Epidemiol 2007;166:1159Y1173 30. Mayne ST, Cartmel B, Kirsh V, et al. Alcohol and tobacco use prediagnosis and postdiagnosis, and survival in a cohort of patients with early stage cancers of the oral cavity, pharynx, and larynx. Cancer Epidemiol Biomarkers Prev 2009;18:3368Y3374 31. Seitz HK, Stickel F. Molecular mechanisms of alcohol-mediated carcinogenesis. Nat Rev Cancer 2007;7:599Y612 32. Wang XD. Alcohol, vitamin A, and cancer. Alcohol 2005;35:251Y258 33. D’Souza G, Kreimer AR, Viscidi R, et al. Case-control study of human papillomavirus and oropharyngeal cancer. N Engl J Med 2007;356: 1944Y1956 34. Bisht M, Bist SS. Human papilloma virus: a new risk factor in a subset of head and neck cancers. J Cancer Res Ther 2011;7:251Y255
Cyclooxygenase-2 Polymorphism and HNSCC
35. Williams CS, Mann M, DuBois RN. The role of cyclooxygenases in inflammation, cancer, and development. Oncogene 1999;18:7908Y7916 36. Wu T. Cyclooxygenase-2 and prostaglandin signaling in cholangiocarcinoma. Biochim Biophys Acta 2005;1755:135Y150 37. Mittal M, Kapoor V, Mohanti BK, et al. Functional variants of COX-2 and risk of tobacco-related oral squamous cell carcinoma in high-risk Asian Indians. Oral Oncol 2010;46:622Y626 38. Zhang X, Miao X, Tan W, et al. Identification of functional genetic variants in cyclooxygenase-2 and their association with risk of esophageal cancer. Gastroenterology 2005;129:565Y576 39. Cok SJ, Morrison AR. The 3’-untranslated region of murine cyclooxygenase-2 contains multiple regulatory elements that alter message stability and translational efficiency. J Biol Chem 2001;276:23179Y23185 40. Kuersten S, Goodwin EB. The power of the 3’ UTR: translational control and development. Nat Rev Genet 2003;4:626Y637 41. Shi J, Misso NL, Kedda MA, et al. Cyclooxygenase-2 gene polymorphisms in an Australian population: association of the -1195G 9A promoter polymorphism with mild asthma. Clin Exp Allergy 2008;38:913Y920 42. Sakaki M, Makino R, Hiroishi K, et al. Cyclooxygenase-2 gene promoter polymorphisms affect susceptibility to hepatitis C virus infection and disease progression. Hepatol Res 2010;40:1219Y1226
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Association Between Cyclooxygenase-2 Gene Polymorphisms and Head and Neck Squamous Cell Carcinoma Risk Yuming Niu, MD,* Hua Yuan, MD,Þ Ming Shen, MD,Þ Huizhang Li, MD,þ Yuanyuan Hu, MD,* and Ning Chen, PhDÞ Abstract: Cyclooxygenase-2 (COX-2) is an inducible enzyme that catalyzes prostaglandins through inflammatory response, which may be involved in autoimmune diseases and cancer pathogenesis. Two potentially functional genetic single-nucleotide polymorphisms (COX-2 -1195G9A and 8473T9C) were supposed to contribute to head and neck squamous cell carcinoma (HNSCC) susceptibility. The aim of this study was to determine the association of these polymorphisms with HNSCC susceptibility in a Chinese Han population. In this study, 2 SNPs were genotyped by TaqMan methods in a patient-control study including 260 patients with HNSCC and 1047 cancer-free controls in a Chinese Han population. We found significant difference in the frequency of alcohol consumption between the patients with HNSCC and controls (P G 0.001), but the genotype frequencies of the 2 polymorphisms were not significantly different between the patients and controls. Further stratified analysis indicated that none of the genotypes were associated with increased risk for HNSCC. This research indicated that the COX-2 -1195G9A and 8473T9C polymorphisms may not be involved in the development of HNSCC in the Chinese Han population. However, further perspective studies are warranted to test these findings and further investigate the potential interactions involving the COX-2 polymorphism and HNSCC.
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From the *Department of Stomatology, Taihe Hospital, Hubei University of Medicine, Shiyan; †Institute of Dental Research, and ‡Department of Epidemiology and Biostatistics, School of Public Health, Nanjing Medical University, Nanjing, People’s Republic of China. Received November 16, 2012. Accepted for publication August 26, 2013. Address correspondence and reprint requests to Ning Chen, PhD, Institute of Dental Research, Nanjing Medical University, 140 Hanzhong Rd, Nanjing, 210029, People’s Republic of China; E-mail: [email protected] Supported by grants from the Foundation of Taihe Hospital of Hubei University of Medicine (2011QD05, 2010D22) and the Medical Development Foundation of Health Department of Jiangsu Province (H200811). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the article. The authors report no conflicts of interest. Copyright * 2014 by Mutaz B. Habal, MD ISSN: 1049-2275 DOI: 10.1097/SCS.0000000000000372
The Journal of Craniofacial Surgery
Key Words: COX-2, polymorphism, head and neck squamous cell carcinoma, susceptibility (J Craniofac Surg 2014;25: 333Y337)
H
ead and neck squamous cell carcinoma (HNSCC) encompasses epithelial malignancies that arise in the paranasal sinuses, nasal cavity, oral cavity, pharynx, and larynx.1 It is the sixth most common malignancy worldwide, which represents approximately 6% of all cases and accounts for approximately 650,000 new cancer cases and 350,000 cancer deaths every year.2,3 Tobacco and alcohol consumption and other environmental sources have been implicated as the important causes of HNSCC development.4,5 Inflammation is an important process in the development of cancer. Prostaglandins are believed to play a key role in inflammation and contribute to cancer development and progression through cellular proliferation and angiogenesis.6,7 Cyclooxygenase (COX, also known as prostaglandin endoperoxide synthases) is a rate-limiting enzyme in conversion of arachidonic acid into prostaglandins. The COX family is composed of 2 isozymes: COX-1 and COX-2.8 Cyclooxygenase-1 is always constitutively expressed and involved in normal physiological functions, whereas COX-2 is an inducible enzyme that catalyzes prostaglandins via inflammatory responses, growth factors, cytokines, and various carcinogens.9,10 Overexpression of COX-2 may be associated with cellular proliferation, inhibition of apoptosis, enhanced invasiveness of cancer cells, and a promotion of angiogenesis11,12 and has been observed in many cancers, especially in the upper aerodigestive tract cancers, such as oral cancer, gastric cancer, and esophageal cancer.13Y15 Some COX-2 inhibitors (eg, nonsteroidal anti-inflammatory drugs, nonsteroidal anti-inflammatory drugs), such as aspirin, indomethacin, and NS398, could inhibit the proliferation of cancer cells independently through the reduction of COX-2 protein expression and suppress the invasiveness of squamous cell carcinoma cell lines via downregulation of matrix metalloproteinase-2 production and activation.16,17 Environmental and genetic risk factors are known to be responsible for HNSCC and act either independently or in combination.3 Interindividual variations in the COX-2 gene may influence the expression level and activity of the protein and subsequently change the susceptibility to developing HNSCC. Two potentially functional single-nucleotide polymorphisms (SNPs), -1195G9A (rs689466) and 8473T9C (rs5275), are located in the promoter region and 3’ untranslated region (UTR) of the COX-2 gene, respectively. These SNPs could affect gene transcription and messenger RNA (mRNA) stability. Lots of molecular epidemiological studies have been conducted to examine the association between these polymorphisms and the susceptibility to HNSCC in diverse populations, but the results remain controversial.18Y23 In this study, we conducted a patientcontrol study to investigate the frequencies of SNPs of COX-2
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between patients with HNSCC and controls in the Chinese Han population and evaluate their contribution to the risk for developing HNSCC.
MATERIALS AND METHODS Study Participants The study was approved by the institutional review board of the Nanjing Medical University, Nanjing, China. All of the participants were genetically unrelated ethnic Han Chinese in Jiangsu province. No restrictions in age, sex, and histology were set, and exclusion criteria were as follows: previous cancer, metastasized cancer, and previous radiotherapy or chemotherapy. A total of 260 patients with newly diagnosed and histologically confirmed squamous cell carcinoma (SCC) of the oral (including the oral cavity and oropharynx) and larynx were recruited from 2008 to 2011 at the Jiangsu Provincial Stomatological Hospital, the Cancer Hospital of Jiangsu Province, and the First Affiliated Hospital of Nanjing Medical University, Nanjing, China. In addition, 1047 cancer-free controls were randomly selected from a pool of 30,000 individuals who participated in a communitybased screening program for noninfectious diseases conducted in Jiangsu province during the same period when the patients were recruited. The control participants had no history of cancer and were frequency matched to the patients by age (T5 y), sex, and residential area (urban or rural). After written informed consent had been obtained, face-to-face interviews were conducted to obtain demographic data (eg, age and sex) and exposure information (eg, smoking status) by trained interviewers. Approximately 5 mL of venous blood sample was collected from each participant. Individuals who smoked 1 cigarette per day for 1 year were defined as ever smokers; otherwise, they were considered as never smokers. In addition, those who consumed 3 or more alcoholic drinks a week for more than 6 months were considered alcohol drinkers.24
Genotyping Genomic DNA was extracted from a leukocyte pellet by proteinase K digestion and followed by phenol-chloroform extraction and ethanol precipitation. All SNPs were analyzed by the conventional TaqMan minor groove binder method. The primers and probes for -1195G9A are as follows: primer: sense, 5’-GCACTACCCATGATAGATGTTAAACAA-3’, antisense, 5’-GGAATTAATTAGATGGAAGGGAGATTT-3’; probe: allele A, FAM-ATGAAATTCCAACTGTC-MGB, allele G, HEX-TGAAAT TCCAGCTGTC-MGB. The primers and probes for 8473T9C are as follows: primer: sense, 5’-TTCCAATGCATCTTCCATGATG-3’, antisense, 5’-ATGCACTGATACCTGTTTTTGTTTG-3’; probe: allele T, FAM-AGTACTTTTGGTTATTTT-MGB, allele C, HEX-AGTA CTTTTGGTCATTTT-MGB. Genotyping was performed without
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TABLE 1. Characteristic Information Between the Patients With HNSCC and Controls Variables Age Mean (SD) e60 960 Sex Male Female Smoking status Never Ever Drinking status Never Ever Tumor location Oral cancer Laryngeal cancer
Patients, n (%)
Controls, n (%)
P
60.41 (8.23) 146 (56.16) 114 (43.84)
59.57 (11.13) 549 (52.44) 498 (47.56)
0.282
180 (69.23) 80 (30.77)
771 (73.64) 276 (26.36)
0.153
134 (51.54) 126 (48.46)
503 (48.04) 544 (51.96)
0.313
135 (51.92) 125 (48.08)
681 (68.04) 366 (34.96)
G0.001
169 (65.00) 91 (35.00)
knowing the patient’s case or control’s status. Ten percent of samples were randomly selected to repeat to validate genotype identification, and the results were totally concordant.
Statistical Analysis The differences in the distributions of demographic characteristics, selected variables, and genotypes of the COX-2 variants between the patients and controls were evaluated with Student’s t-test or Chi-squared tests. The Hardy-Weinberg equilibrium of the genotype distribution of the controls was tested by a goodness-of-fit Chi-squared test. Linkage disequilibrium was estimated with an online software platform.25 The associations between COX-2 SNPs and HNSCC risk were estimated by computing the odds ratios (ORs) and their 95% confidence intervals (CIs) from both univariate and multivariate logistic regression analyses without and with the adjustments for age, sex, smoking status, and drinking status. All statistical analyses were performed with SAS 9.1.3 software (SAS Institute, Cary, NC). Statistical significance was set at P G 0.05, and all statistical tests were 2-sided.
RESULTS The demographic characteristics and clinical information for the 260 patients with HNSCC (mean [SD] age, 60.41 [8.23]) and 1047 controls (mean [SD] age, 59.57 [11.13]) enrolled in our study are shown in Table 1. Approximately 48.46% of the patients with
TABLE 2. Odds Ratios and Confidence Intervals of Associations Between COX-2 SNPs and HNSCC (Oral SCC and Laryngeal SCC)
Genotypes
Controls, n (%)
All Patients, n (%)
-1195G9A GG GA AA GA+AA 8473T9C TT TC CC TC+CC
1035 222 (21.45) 542 (52.37) 271 (26.18) 813 (78.55) 1032 691 (66.96) 316 (30.62) 25 (2.42) 341 (33.04)
259 (23.55) (48.65) (27.80) (76.45) 258 177 (68.60) 72 (27.91) 9 (3.49) 81 (31.40) 61 126 72 198
Adjusted OR (95% CI)*
1.00 0.85 (0.60Y1.21) 1.01 (0.69Y1.50) 0.91 (0.65Y1.26) 1.00 0.90 (0.66Y1.22) 1.48 (0.68Y3.25) 0.94 (0.70Y1.26)
Oral SCC, n (%) 169 (26.04) (47.34) (26.63) (73.96) 168 118 (70.24) 45 (26.79) 5 (2.98) 50 (29.76)
44 80 45 125
Adjusted OR (95% CI)*
1.00 0.74 (0.49Y1.11) 0.87 (0.55Y1.39) 0.78 (0.53Y1,14) 1.00 0.86 (0.58Y1.26) 1.03 (0.36Y2.97) 0.87 (0.60Y1.27)
Laryngeal SCC, n (%) 90 (18.89) (51.11) (30.00) (81.11) 90 59 (65.56) 27 (30.00) 4 (4.44) 31 (34.44) 17 46 27 73
Adjusted OR (95% CI)*
1.00 1.16 (0.65Y2.09) 1.43 (0.75Y2.75) 1.23 (0.71Y2.15) 1.00 1.02 (0.63Y1.64) 1.62 (0.54Y4.88) 1.07 (0.67Y1.69)
*Adjusted by age, sex, smoking status, and drinking status.
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The Journal of Craniofacial Surgery
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Cyclooxygenase-2 Polymorphism and HNSCC
TABLE 3. Stratified Analysis on the Association Between COX-2 SNPs and HNSCC Risk -1195G9A Patient (n = 259) Characteristics Age, y e60 960 Sex Male Female Smoking status Never Ever Drinking status Never Ever
8473T9C
Control (n = 1035)
Patient (n = 258)
Control (n = 1032)
GG
GA+AA
GG
GA+AA
OR (95% CI)*
TT
TC+CC
TT
TC+CC
OR (95% CI)*
36 (24.83) 25 (21.93)
109 (75.17) 89 (78.07)
114 (20.99) 108 (21.95)
429 (79.01) 384 (78.05)
0.85 (0.55,1.32) 1.01 (0.61, 1.67)
99 (68.28) 78 (69.03)
46 (31.72) 35 (30.97)
375 (69.19) 316 (64.49)
167 (30.81) 174 (35.51)
1.08 (0.72, 1.61) 0.80 (0.51, 1.25)
40 (22.35) 21 (26.25)
139 (77.65) 59 (73.75)
161 (21.18) 61 (22.18)
599 (78.82) 214 (77.82)
0.99 (0.66, 1.49) 0.87 (0.48, 1.58)
124 (68.89) 53 (67.95)
56 (31.11) 25 (32.05)
509 (67.06) 182 (66.67)
250 (32.94) 91 (33.33)
0.96 (0.67, 1.37) 1.10 (0.63, 1.92)
21 (22.58) 40 (24.10)
72 (77.42) 126 (75.90)
104 (20.88) 118 (21.97)
394 (79.12) 419 (78.03)
1.05 (0.60, 1.81) 0.94 (0.62, 1.44)
63 (69.23) 114 (68.26)
28 (30.77) 53 (31.74)
333 (67.00) 358 (66.92)
164 (33.00) 177 (33.08)
0.95 (0.57, 1.56) 1.04 (0.71, 1.53)
21 (21.65) 40 (24.69)
76 (78.35) 122 (75.31)
111 (22.98) 111 (20.11)
372 (77.02) 441 (79.89)
1.15 (0.67, 1.98) 0.87 (0.57, 1.34)
66 (69.47) 111 (68.10)
29 (30.53) 52 (31.90)
319 (66.18) 372 (67.64)
163 (33.82) 178 (32.36)
0.90 (0.55, 1.48) 1.06 (0.72, 1.57)
*Adjusted by age, sex, smoking status, and drinking status.
HNSCC were smokers, which is not significantly different from that of the controls (51.96%) (P 9 0.05); otherwise, the HNSCC group had significantly more drinkers (48.08% versus 34.96%, P G 0.001) than those in the control group. The genotype distributions of the 2 SNPs in the patients and controls are shown in Table 2. The observed genotype frequencies for the 2 polymorphisms in the controls were all in Hardy-Weinberg equilibrium (P = 0.110 and 0.112 for -1195G9A and 8473T9C, respectively), which indicates good homogeneity within the study participants. One patient (0.38%) and 12 controls (1.15%) for -1195G9A as well as 2 patients (0.77%) and 12 controls (1.24%) for 8473T9C were not able to be genotyped owing to poor DNA quantity and/or quality. For -1195G9A, the genotype frequencies of GG, GA, and AA in the controls were 21.45%, 52.37%, and 26.18%, respectively, whereas those of the patients with HNSCC were 23.55%, 48.65%, and 27.80%, respectively (P = 0.555). Similarly, no statistically significant differences among the genotype frequencies of 8473T9C were observed (P = 0.478). In the multivariate logistic regression analysis, neither the heterozygote (for -1195G9A GA genotype and 8473T9C TC genotype) nor the mutated homozygote (for -1195G9A AA genotype and 8473T9C CC genotype) of the SNPs was associated with the developing risk for HNSCC, compared with their corresponding wild homozygote (for -1195G9A GG genotype and 8473T9C TT genotype). When the heterozygote and mutated homozygote were combined as the dominant model (for -1195G9A GA/AA genotype and 8473T9C TC/CC genotype), nonsignificant associations were observed between the variant genotypes and HNSCC risk (Table 2). In addition, further stratified analysis of -1195G9A and 8473T9C suggested no significant association with HNSCC after adjusting for age, sex, smoking status, and drinking status (Table 3). The estimation of linkage disequilibrium showed that -1195G9A and 8473T9C were not in strong linkage disequilibrium with the standardized D’= 0.978 and r2 = 0.189. We thus considered the 2 SNPs together but found no significant association for HNSCC with the combined COX-2 genotypes (Table 4).
DISCUSSION Epidemiological studies have demonstrated that the development of HNSCC may be largely caused by alcohol and tobacco use.5,26 An increase of approximately 6-fold in HNSCC risk has been
observed among smokers compared with nonsmokers.3,27 Furthermore, the risk for developing HNSCC apparently increases if smokers are also heavy drinkers,4,28 although our research showed that characteristic information does not indicate significant associations between smoking and HNSCC. On the other hand, an almost 2-fold increased risk for HNSCC has been observed in drinkers. As we know, acetaldehyde can generate from ethanol through oxidation by alcohol dehydrogenase, which is a strong risk factor for the development of HNSCC.29,30 Alcohol dehydrogenase can induce gene mutations, especially inactive variant aldehyde dehydrogenase-2 genotype to increase the accumulation of acetaldehyde after alcohol consumption.31,32 Furthermore, some new evidences also indicate that the human papilloma virus infection, especially that by human papilloma virus type 16, is a risk factor for HNSCC.33,34 The COX-2 gene that encodes the COX-2 enzyme has been mapped to chromosome 1q25.2Y25.3. Cyclooxygenase-2 is a key enzyme produced during the conversion of free arachidonic acid into prostaglandins, which is relevant to cancer development and progression.35 Furthermore, the increased expression of COX-2 could be involved in tumorigenesis by enhancing tumor cell proliferation, angiogenesis, inhibition of apoptosis, immune escape, and increased metastasis.36 Meanwhile, the expression and stability of COX-2 mRNA will be regulated by various elements of the 5’ promoter region and 3’UTR of the transcript.37 The SNP -1195G9A, which is located in the promoter region, may create a c-MYB binding site with strikingly high promoter activity.38 Another SNP located in the 3’UTR, 8473T9C, will alter mRNA stability as well as translation efficiency and subsequently influences cancers susceptibility.39,40
TABLE 4. Locus-Locus Interaction Between -1195G9A and 8473T9C on HNSCC Risk Genotypes
Patient -1195G9A 8473T9C (n = 257)
GG GG GA+AA GA+AA
Control (n = 1029)
OR (95 CI%)
OR (95 CI%)*
TT 60 (23.35) 220 (21.38) 1.00 1.00 TC+CC 1 (0.39) 2 (0.19) 1.84 (0.16Y20.5) 1.67 (0.14Y19.45) TT 116 (45.14) 468 (45.48) 0.91 (0.64Y1.29) 0.93 (0.65Y1.32) TC+CC 80 (31.13) 339 (32.94) 0.87 (.060Y1.26) 0.89 (.061Y1.30)
*Adjusted by age, sex, smoking status, and drinking status.
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Chiang et al21 recently conducted a patient-control study of 377 patients with oral SCC and 442 controls in a China Taiwan population and found that COX-2 -1195AA genotypes are associated with a significant 1.55-fold (95% confidence interval [CI], 1.04Y2.31) increase in risk for developing oral cancer compared with their wildtype genotypes. However, patient-control studies in a white and Indian populations showed that the -1195G9A polymorphism is not significantly associated with HNSCC risk.19,37 Moreover, using dual-luciferase reporter analysis with plasmids containing the 1195G9A SNP to analyze transcription activity, Shi et al41 and Sakaki et al42 reported that the promoter region containing -1195G shows significantly higher transcription activity than that containing 1195A. However, a study performed by Zhang et al38 suggested opposing findings. In molecular epidemiological studies on the association between COX-2 8473T9C polymorphism and cancer susceptibility, results conflicted among different types of cancer and no significant association with HNSCC was found.23,37 This patient-control study comprised 260 patients with HNSCC and 1047 healthy controls in the Chinese Han population. Similar frequencies of the alleles and genotypes were found, and none of the genotypes were associated with increased risks for HNSCC, which suggests that no genotype contributes to the development of HNSCC in the Chinese Han population. To improve the power of genetic analysis and understand the results of risk assessment, we stratified all patients with HNSCC into 2 subgroups (oral cancer and laryngeal cancer). Further stratified analyses (age, sex, smoking status, drinking status, and haplotype frequency assessments) were also performed. The genotype frequency was similarly distributed between each of the subgroups and the controls. None of the risk genotypes were found to be in association with any subgroup of HNSCC. Several limitations in our study must be addressed. First, other SNP loci, such as -765 G9C in the promoter region of the COX-2 gene, have been related to cancer risk. Unfortunately, because of technical reasons, we were unable to detect the genotype of the -765 G9C polymorphism in the patient and control groups successfully. The association between -765 G9C polymorphism and HNSCC risk as well as the combined effect of the haplotype of -765 G9C and -1195G9A polymorphism in the promoter region were not observed in this research. Second, the sample size of the patients with HNSCC was not sufficiently large and information from some patients was not collected completely during the early stage of this research. The association between the -1195G9A polymorphism and cancer subtypes or tumor stages was not explored. This limitation could reduce the validity of the research findings because some potentially hidden risk factors may not have been fully exposed. Third, inherent biases may have led to spurious findings because the patients were recruited from the hospital and the controls were selected from a similar population. Despite these limitations, the potential bias was considered minimal and had no influence on the results because the controls were matched to the patients with HNSCC on the basis of age, sex, as well as residential area and drinking and smoking statuses were adjusted during the statistical analyses. In conclusion, the current study did not find any association between the 2 functional genetic variants of the COX-2 gene and HNSCC risk in the Chinese Han population. Well-designed studies are further warranted to validate these findings and investigate the potential gene-gene and gene-environment interactions involved in COX-2 polymorphisms and HNSCC.
REFERENCES 1. Argiris A, Eng C. Epidemiology, staging, and screening of head and neck cancer. Cancer Treat Res 2003;114:15Y60 2. Parkin DM, Bray F, Ferlay J, et al. Global cancer statistics, 2002. CA Cancer J Clin 2005;55:74Y108
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3. Argiris A, Karamouzis MV, Raben D, et al. Head and neck cancer. Lancet 2008;371:1695Y1709 4. Blot WJ, McLaughlin JK, Winn DM, et al. Smoking and drinking in relation to oral and pharyngeal cancer. Cancer Res 1988;48:3282Y3287 5. Vineis P, Alavanja M, Buffler P, et al. Tobacco and cancer: recent epidemiological evidence. J Natl Cancer Inst 2004;96:99Y106 6. Cao Y, Prescott SM. Many actions of cyclooxygenase-2 in cellular dynamics and in cancer. J Cell Physiol 2002;190:279Y286 7. Romano M, Claria J. Cyclooxygenase-2 and 5-lipoxygenase converging functions on cell proliferation and tumor angiogenesis: implications for cancer therapy. FASEB J 2003;17:1986Y1995 8. Simmons DL, Botting RM, Hla T. Cyclooxygenase isozymes: the biology of prostaglandin synthesis and inhibition. Pharmacol Rev 2004;56:387Y437 9. Vane SJ. Differential inhibition of cyclooxygenase isoforms: an explanation of the action of NSAIDs. J Clin Rheumatol 1998;4:s3Ys10 10. Herschman HR. Prostaglandin synthase 2. Biochim Biophys Acta 1996;1299:125Y140 11. Souza RF, Shewmake K, Beer DG, et al. Selective inhibition of cyclooxygenase-2 suppresses growth and induces apoptosis in human esophageal adenocarcinoma cells. Cancer Res 2000;60:5767Y5772 12. Ranger GS, Thomas V, Jewell A, et al. Elevated cyclooxygenase-2 expression correlates with distant metastases in breast cancer. Anticancer Res 2004;24:2349Y2351 13. Bayazit YA, Buyukberber S, Sari I, et al. Cyclo-oxygenase 2 expression in laryngeal squamous cell carcinoma and its clinical correlates. ORL J Otorhinolaryngol Relat Spec 2004;66:65Y69 14. Segawa E, Sakurai K, Kishimoto H, et al. Expression of cyclooxygenase-2 and DNA topoisomerase II alpha in precancerous and cancerous lesions of the oral mucosa. Oral Oncol 2008;44:664Y671 15. Liu X, Li P, Zhang ST, et al. COX-2 mRNA expression in esophageal squamous cell carcinoma (ESCC) and effect by NSAID. Dis Esophagus 2008;21:9Y14 16. Gao J, Niwa K, Sun W, et al. Non-steroidal anti-inflammatory drugs inhibit cellular proliferation and upregulate cyclooxygenase-2 protein expression in endometrial cancer cells. Cancer Sci 2004;95:901Y907 17. Kurihara Y, Hatori M, Ando Y, et al. Inhibition of cyclooxygenase-2 suppresses the invasiveness of oral squamous cell carcinoma cell lines via down-regulation of matrix metalloproteinase-2 production and activation. Clin Exp Metastasis 2009;26:425Y432 18. Jeong SW, Tae K, Lee SH, et al. Cox-2 and IL-10 polymorphisms and association with squamous cell carcinoma of the head and neck in a Korean sample. J Korean Med Sci 2010;25:1024Y1028 19. Peters WH, Lacko M, Te Morsche RH, et al. COX-2 polymorphisms and the risk for head and neck cancer in white patients. Head Neck 2009;31:938Y943 20. Pu X, Lippman SM, Yang H, et al. Cyclooxygenase-2 gene polymorphisms reduce the risk of oral premalignant lesions. Cancer 2009;115:1498Y1506 21. Chiang SL, Chen PH, Lee CH, et al. Up-regulation of inflammatory signalings by areca nut extract and role of cyclooxygenase-2 -1195G9A polymorphism reveal risk of oral cancer. Cancer Res 2008;68: 8489Y8498 22. Lin YC, Huang HI, Wang LH, et al. Polymorphisms of COX-2 -765G9C and p53 codon 72 and risks of oral squamous cell carcinoma in a Taiwan population. Oral Oncol 2008;44:798Y804 23. Campa D, Hashibe M, Zaridze D, et al. Association of common polymorphisms in inflammatory genes with risk of developing cancers of the upper aerodigestive tract. Cancer Causes Control 2007;18:449Y455 24. Chu H, Cao W, Chen W, et al. Potentially functional polymorphisms in IL-23R and risk of esophageal cancer in a Chinese population. Int J Cancer 2011;130:1093Y1097 25. Shi YY, He L. SHEsis, a powerful software platform for analyses of linkage disequilibrium, haplotype construction, and genetic association at polymorphism loci. Cell Res 2005;15:97Y98 26. Secretan B, Straif K, Baan R, et al. A review of human carcinogensVPart E: tobacco, areca nut, alcohol, coal smoke, and salted fish. Lancet Oncol 2009;10:1033Y1034
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27. Kamangar F, Dores GM, Anderson WF. Patterns of cancer incidence, mortality, and prevalence across five continents: defining priorities to reduce cancer disparities in different geographic regions of the world. J Clin Oncol 2006;24:2137Y2150 28. Smith EM, Rubenstein LM, Haugen TH, et al. Tobacco and alcohol use increases the risk of both HPV-associated and HPV-independent head and neck cancers. Cancer Causes Control 2010;21:1369Y1378 29. Guha N, Boffetta P, Wunsch Filho V, et al. Oral health and risk of squamous cell carcinoma of the head and neck and esophagus: results of two multicentric case-control studies. Am J Epidemiol 2007;166:1159Y1173 30. Mayne ST, Cartmel B, Kirsh V, et al. Alcohol and tobacco use prediagnosis and postdiagnosis, and survival in a cohort of patients with early stage cancers of the oral cavity, pharynx, and larynx. Cancer Epidemiol Biomarkers Prev 2009;18:3368Y3374 31. Seitz HK, Stickel F. Molecular mechanisms of alcohol-mediated carcinogenesis. Nat Rev Cancer 2007;7:599Y612 32. Wang XD. Alcohol, vitamin A, and cancer. Alcohol 2005;35:251Y258 33. D’Souza G, Kreimer AR, Viscidi R, et al. Case-control study of human papillomavirus and oropharyngeal cancer. N Engl J Med 2007;356: 1944Y1956 34. Bisht M, Bist SS. Human papilloma virus: a new risk factor in a subset of head and neck cancers. J Cancer Res Ther 2011;7:251Y255
Cyclooxygenase-2 Polymorphism and HNSCC
35. Williams CS, Mann M, DuBois RN. The role of cyclooxygenases in inflammation, cancer, and development. Oncogene 1999;18:7908Y7916 36. Wu T. Cyclooxygenase-2 and prostaglandin signaling in cholangiocarcinoma. Biochim Biophys Acta 2005;1755:135Y150 37. Mittal M, Kapoor V, Mohanti BK, et al. Functional variants of COX-2 and risk of tobacco-related oral squamous cell carcinoma in high-risk Asian Indians. Oral Oncol 2010;46:622Y626 38. Zhang X, Miao X, Tan W, et al. Identification of functional genetic variants in cyclooxygenase-2 and their association with risk of esophageal cancer. Gastroenterology 2005;129:565Y576 39. Cok SJ, Morrison AR. The 3’-untranslated region of murine cyclooxygenase-2 contains multiple regulatory elements that alter message stability and translational efficiency. J Biol Chem 2001;276:23179Y23185 40. Kuersten S, Goodwin EB. The power of the 3’ UTR: translational control and development. Nat Rev Genet 2003;4:626Y637 41. Shi J, Misso NL, Kedda MA, et al. Cyclooxygenase-2 gene polymorphisms in an Australian population: association of the -1195G 9A promoter polymorphism with mild asthma. Clin Exp Allergy 2008;38:913Y920 42. Sakaki M, Makino R, Hiroishi K, et al. Cyclooxygenase-2 gene promoter polymorphisms affect susceptibility to hepatitis C virus infection and disease progression. Hepatol Res 2010;40:1219Y1226
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