Tumor Biol. DOI 10.1007/s13277-015-3334-1

RESEARCH ARTICLE

Clinical significance of serum expression of GROβ in hepatocellular carcinoma YueGuo Li 1,3 & YuFang Wang 2 & Peng Zhang 1,3

Received: 9 January 2015 / Accepted: 12 March 2015 # International Society of Oncology and BioMarkers (ISOBM) 2015

Abstract This study aimed to determine whether serum levels of growth-related gene product β (GROβ) were associated with clinical parameters in hepatocellular carcinoma (HCC). Using an enzyme-linked immunosorbent assay (ELISA), the serum GROβ levels of 80 HCC patients, 65 patients with benign diseases of the liver, and 60 healthy volunteers were examined. The association between serum levels of GROβ and clinical parameters of HCC was analyzed statistically. The serum GROβ levels were much lower in benign diseases and healthy volunteers than HCC, and associated with tumor node metastasis (TNM) stages, tumor size, vascular thrombosis, capsule, and Edmondson grading of HCC (p0.05). We have demonstrated that GROβ, as an oncogene product, contributed to tumorigenesis and metastasis of HCC.

Keywords HCC . GROβ . Serum . Tumor

* Peng Zhang [email protected]

Introduction CXC chemokine receptors are integral membrane proteins that specifically bind and are activated by cytokines of the CXC chemokine family. CXCL2 chemokine, also known as growth-related gene product β (GROβ), belongs to the CXC chemokine group. GROβ belongs to growth-related gene product (GRO), which is composed of GROα, GROβ, and GROγ [1, 2]. GROα is highly expressed in a variety of tumors and plays an important role in tumor proliferation [3], angiogenesis [4], and metastasis [5]. Compared with GROα, the roles of GROβ in tumors are poorly understood. Primary hepatocellular carcinoma (HCC) is one of the most lethal malignancies worldwide. It is the third leading cause of cancer death in China. Most cases of HCC arise from chronic infection with human hepatitis B virus (HBV) or human hepatitis C virus (HCV). We previously studied that CCL15, known as important chemokine, might be a specific proteomic biomarker of HCC, which had an important role in tumorigenesis and tumor invasion [6]. We also previously observed that GROβ was highly expressed in HCC tissues and cell lines (data unpublished). Although this suggests that GROβ may be secreted from tumor tissues into the extracellular milieu, no report has yet investigated serum GROβ expression levels in human HCC. In this study, we measured the serum levels of GROβ in HCC patients and healthy controls by an enzyme-linked immunosorbent assay (ELISA) and analyzed the association between clinical parameters of HCC and serum GROβ levels.

1

Department of Laboratory, National Clinical Research Center of Cancer, Tianjin Medical University Cancer Institute and Hospital, Tianjin 300060, People’s Republic of China

2

HeBei United University Affiliated Hospital, Hebei 063000, People’s Republic of China

Materials and methods

Key Laboratory of Cancer Prevention and Therapy, The National B863^ Program of Clinical Research Laboratory, Tianjin 300060, People’s Republic of China

The study was approved by the Research Ethics Committee of Tianjin Medical University Cancer Institute and Hospital.

3

Tumor Biol.

Informed consent was obtained from all of the patients. All specimens were handled and made anonymous according to ethical and legal standards. Patients with HCC Serum samples were collected from 80 HCC patients from Tianjin Medical University Cancer Institute and Hospital from July 2009 to August 2010 (50 males and 30 females; mean age 61; range 41–76 years). Table 1 summarizes the clinicopathological data of these patients with HCC. Control patients Serum samples were obtained from 65 patients with benign diseases of the liver from Tianjin Medical University Cancer Table 1 Clinicopathological data of patients with HCC, benign diseases, and healthy controls

Variance

HCC patients (n=80)

Institute and Hospital from July 2009 to August 2010. The benign diseases are as follows: liver cirrhosis (n=25; mean age 61 years), calculus of bile duct (n=20; mean 60 years); hepatitis B (n=20; mean age 59 years) (Table 1). In addition, 60 blood donors (mean age 61 years) without liver neoplasia, alcoholic cirrhosis, or hepatitis B or hepatitis C infection were recruited from routine health examinations at Tianjin Medical University Cancer Institute and Hospital from July 2009 to August 2010 (Table 1). Measurement of GROβ using ELISA A human GROβ ELISA development kit (PeproTech, Rocky Hill, NJ, USA) was used to measure GROβ levels in human serum; 96-well ELISA microplates were coated overnight with 100 μL of GROβ antibody (PeproTech, Rocky Hill, Benign diseases (n=65)

Healthy controls (n=60)

Liver cirrhosis (n=25)

Calculus of bile duct (n=20)

Hepatitis B (n=20)

12 13

9 11

10 10

32 28

11 14

13 7

12 8

31 29

– –

– –

– –

– –

25 0

– –

– –

– –

– –

– –

– –

– –

– –

– –

– –

– –

– –

– –

– –

– –

– –

– –

– –

– –

– – –

– – –

– – –

– – –

Sex Male 50 Female 30 Age ≥60 25