Tumor Biol. DOI 10.1007/s13277-014-1923-z
RESEARCH ARTICLE
Clinical significance of serum tenascin-c levels in epithelial ovarian cancer Tastekin Didem & Tas Faruk & Karabulut Senem & Duranyildiz Derya & Serilmez Murat & Guveli Murat & Kaynur Oznur
Received: 22 January 2014 / Accepted: 1 April 2014 # International Society of Oncology and BioMarkers (ISOBM) 2014
Abstract Tenascin-C (TNC) is an extracellular matrix protein that is expressed at low levels in normal adult tissue but is highly expressed around many tumors including ovarian tumors. The objective of this study was to determine the clinical significance of the serum levels of TNC in epithelial ovarian cancer (EOC) patients. A total of 50 patients with a pathologically confirmed diagnosis of EOC were included in this study. Serum TNC levels were determined by the solidphase sandwich enzyme-linked immunosorbent assay (ELISA) method. Age- and sex- matched 28 healthy controls were included in the analysis. Median age of the patients was 56.5 years old, range 22 to 83 years. Majority of the patients had advanced disease (FIGO stage III–IV) (90 %). The median serum TNC levels were found significantly higher in EOC patients (130.5 pg/mL) compared to healthy controls (90.1 pg/ mL) (p=0.03). We found no correlation between serum TNC levels and any prognostic parameters analyzed, including age of the patients, histology, tumor grade, stage of the disease, and response to chemotherapy. Survival analysis did not show statistically significant effect of serum TNC concentration on progression-free and overall survival (p=0.36 and p=0.19, T. Didem (*) : T. Faruk : K. Senem Department of Medical Oncology, Oncology Institute, Istanbul University, Capa, 34390 Istanbul, Turkey e-mail: [email protected] D. Derya : S. Murat Department of Basic Oncology, Oncology Institute, Istanbul University, Istanbul, Turkey G. Murat Department of Radiation Oncology, Oncology Institute, Istanbul University, Istanbul, Turkey K. Oznur Dr. Abdurrahman Yurtaslan Ankara Oncology Education and Research Hospital, Ankara, Turkey
respectively). However, patients with high serum TNC levels tend to have poor overall survival. In conclusion, although serum TNC levels are elevated, it has no predictive or prognostic roles on survival in EOC patients.
Keywords Tenascin-C . Epithelial ovarian cancer . Prognosis
Introductıon Tenascin-C (TNC) is a key factor in tissue remodeling, and its deregulated high expression is causally linked to many diseases, including heart failure, thrombosis, atherosclerosis, and cancer [1]. High TNC expression has been shown in most solid cancers, including those arising in the brain, breast, uterus, prostate, pancreas, colon, stomach, mouth, larynx, lung, liver, kidney, bladder, skin, bone, soft tissues, lymphomas, and in ovaries [2]. TNC plays a role in enhancing proliferation, invasion, and angiogenesis during tumorigenesis and metastasis. For example, in a mouse model, TNC knockdown causes dramatic inhibition of lung metastasis and colonization by breast cancer cells [3]. Moreover, the data suggest that it contributes to cancer formation via its potential interference with genomic stability, by blocking immunosurveillance and by providing a favorable niche for normal and tumor stem cells to survive and expand [4]. A correlation between high TNC expression and poor survival was demonstrated in some cancer types such as glioma, breast, colon, and lung cancer [2]. Although these available findings were existed in the other tumors, there are very few clinical studies to investigate the role of TNC in ovarian cancer [5, 6]. The present study was designed to assess the clinical significance of serum TNC level in epithelial ovarian cancer (EOC) patients.
Tumor Biol.
Materıal and methods Patients A total of 50 EOC patients with histologically proven diagnosis treated at Istanbul University, Institute of Oncology were enrolled into the study. The staging was established in accordance with International Federation of Gynecologists and Obstetricians (FIGO) classification. Patients with stage III bulky disease or stage IV disease were initially treated with neoadjuvant chemotherapy and operated afterwards. Patients with operable stage III disease who had undergone primary surgery consist of total abdominal hysterectomy, bilateral salpingoopherectomy, appendectomy, omentectomy, and pelvic and/or paraaortic lymphadenectomy were treated with adjuvant chemotherapy. All patients received standard paclitaxel-carboplatin-containing chemotherapy regimen. For comparison of serum levels of TNC, age- and sexmatched 28 healthy women controls were included in the analysis. Institutional review board approval was obtained from each subject prior to the commencement of the study. Measurement of serum TNC levels Serum samples were obtained on the first admission before any adjuvant and metastatic treatment was given or follow-up patients. Blood samples were obtained from EOC patients and healthy controls by venipuncture and clotted at room temperature. The sera were collected following centrifugation and frozen immediately at −20 °C until analysis. Serum TNC levels were determined by the solid-phase sandwich ELISA method. The TNC ELISA (USCN, China) uses a doubleantibody sandwich ELISA to determine the level of TNC in samples. There are two antibodies against human TNC used in kit, so the kit can detect both large and small splice variants of TNC. Serum samples and standards are added to the wells which are precoated with Human TNC monoclonal antibody. Following incubation, TNC antibodies labeled with biotin and combined with streptavidin-HRP are added to form immune complex and allowed to incubate for 1 h. Unbound material is washed away, and then chromogen solution is added for the conversion of the colorless solution to a blue solution, the intensity of which is proportional to the amount of TNC in the sample. As the effect of the acidic stop solution, the color has become yellow. The colored reaction product is measured using an automated ELISA reader (Rayto, RT-1904C Chemistry Analyzer, Atlanta, GA, USA). The results were expressed as pg/mL.
between various clinical/laboratory parameters and serum levels of TNC assays was accomplished using MannWhitney U test. Overall survival (OS) was calculated from the date of first admission to the clinics to disease-related death or date of last contact with the patient or any family member. Progression-free survival (PFS) was calculated from the date of admission to the date of the first radiologic progression with/without elevated serum tumor marker. KaplanMeier method was used for estimation of survival distribution and differences in survival were assessed by the log-rank statistics. A p value
RESEARCH ARTICLE
Clinical significance of serum tenascin-c levels in epithelial ovarian cancer Tastekin Didem & Tas Faruk & Karabulut Senem & Duranyildiz Derya & Serilmez Murat & Guveli Murat & Kaynur Oznur
Received: 22 January 2014 / Accepted: 1 April 2014 # International Society of Oncology and BioMarkers (ISOBM) 2014
Abstract Tenascin-C (TNC) is an extracellular matrix protein that is expressed at low levels in normal adult tissue but is highly expressed around many tumors including ovarian tumors. The objective of this study was to determine the clinical significance of the serum levels of TNC in epithelial ovarian cancer (EOC) patients. A total of 50 patients with a pathologically confirmed diagnosis of EOC were included in this study. Serum TNC levels were determined by the solidphase sandwich enzyme-linked immunosorbent assay (ELISA) method. Age- and sex- matched 28 healthy controls were included in the analysis. Median age of the patients was 56.5 years old, range 22 to 83 years. Majority of the patients had advanced disease (FIGO stage III–IV) (90 %). The median serum TNC levels were found significantly higher in EOC patients (130.5 pg/mL) compared to healthy controls (90.1 pg/ mL) (p=0.03). We found no correlation between serum TNC levels and any prognostic parameters analyzed, including age of the patients, histology, tumor grade, stage of the disease, and response to chemotherapy. Survival analysis did not show statistically significant effect of serum TNC concentration on progression-free and overall survival (p=0.36 and p=0.19, T. Didem (*) : T. Faruk : K. Senem Department of Medical Oncology, Oncology Institute, Istanbul University, Capa, 34390 Istanbul, Turkey e-mail: [email protected] D. Derya : S. Murat Department of Basic Oncology, Oncology Institute, Istanbul University, Istanbul, Turkey G. Murat Department of Radiation Oncology, Oncology Institute, Istanbul University, Istanbul, Turkey K. Oznur Dr. Abdurrahman Yurtaslan Ankara Oncology Education and Research Hospital, Ankara, Turkey
respectively). However, patients with high serum TNC levels tend to have poor overall survival. In conclusion, although serum TNC levels are elevated, it has no predictive or prognostic roles on survival in EOC patients.
Keywords Tenascin-C . Epithelial ovarian cancer . Prognosis
Introductıon Tenascin-C (TNC) is a key factor in tissue remodeling, and its deregulated high expression is causally linked to many diseases, including heart failure, thrombosis, atherosclerosis, and cancer [1]. High TNC expression has been shown in most solid cancers, including those arising in the brain, breast, uterus, prostate, pancreas, colon, stomach, mouth, larynx, lung, liver, kidney, bladder, skin, bone, soft tissues, lymphomas, and in ovaries [2]. TNC plays a role in enhancing proliferation, invasion, and angiogenesis during tumorigenesis and metastasis. For example, in a mouse model, TNC knockdown causes dramatic inhibition of lung metastasis and colonization by breast cancer cells [3]. Moreover, the data suggest that it contributes to cancer formation via its potential interference with genomic stability, by blocking immunosurveillance and by providing a favorable niche for normal and tumor stem cells to survive and expand [4]. A correlation between high TNC expression and poor survival was demonstrated in some cancer types such as glioma, breast, colon, and lung cancer [2]. Although these available findings were existed in the other tumors, there are very few clinical studies to investigate the role of TNC in ovarian cancer [5, 6]. The present study was designed to assess the clinical significance of serum TNC level in epithelial ovarian cancer (EOC) patients.
Tumor Biol.
Materıal and methods Patients A total of 50 EOC patients with histologically proven diagnosis treated at Istanbul University, Institute of Oncology were enrolled into the study. The staging was established in accordance with International Federation of Gynecologists and Obstetricians (FIGO) classification. Patients with stage III bulky disease or stage IV disease were initially treated with neoadjuvant chemotherapy and operated afterwards. Patients with operable stage III disease who had undergone primary surgery consist of total abdominal hysterectomy, bilateral salpingoopherectomy, appendectomy, omentectomy, and pelvic and/or paraaortic lymphadenectomy were treated with adjuvant chemotherapy. All patients received standard paclitaxel-carboplatin-containing chemotherapy regimen. For comparison of serum levels of TNC, age- and sexmatched 28 healthy women controls were included in the analysis. Institutional review board approval was obtained from each subject prior to the commencement of the study. Measurement of serum TNC levels Serum samples were obtained on the first admission before any adjuvant and metastatic treatment was given or follow-up patients. Blood samples were obtained from EOC patients and healthy controls by venipuncture and clotted at room temperature. The sera were collected following centrifugation and frozen immediately at −20 °C until analysis. Serum TNC levels were determined by the solid-phase sandwich ELISA method. The TNC ELISA (USCN, China) uses a doubleantibody sandwich ELISA to determine the level of TNC in samples. There are two antibodies against human TNC used in kit, so the kit can detect both large and small splice variants of TNC. Serum samples and standards are added to the wells which are precoated with Human TNC monoclonal antibody. Following incubation, TNC antibodies labeled with biotin and combined with streptavidin-HRP are added to form immune complex and allowed to incubate for 1 h. Unbound material is washed away, and then chromogen solution is added for the conversion of the colorless solution to a blue solution, the intensity of which is proportional to the amount of TNC in the sample. As the effect of the acidic stop solution, the color has become yellow. The colored reaction product is measured using an automated ELISA reader (Rayto, RT-1904C Chemistry Analyzer, Atlanta, GA, USA). The results were expressed as pg/mL.
between various clinical/laboratory parameters and serum levels of TNC assays was accomplished using MannWhitney U test. Overall survival (OS) was calculated from the date of first admission to the clinics to disease-related death or date of last contact with the patient or any family member. Progression-free survival (PFS) was calculated from the date of admission to the date of the first radiologic progression with/without elevated serum tumor marker. KaplanMeier method was used for estimation of survival distribution and differences in survival were assessed by the log-rank statistics. A p value
