Impact of Hepatitis C Virus (HCV) infection on biomolecular markers influencing the pathogenesis of bladder cancer

Abstract

Objective: The present study was designed to determine the possible impact of hepatitis C virus (HCV) infection on the expression of telomerase (TERT), retinoblastoma (RB1), E2F3, TP53, CDKN1A (p21) and fibroblast growth factor receptor- 3 (FGFR3) genes in patients with bladder cancer (BC).

Materials and methods: 100 patients with bladder cancer (15 female and 85 male) were divided into 2 groups; Group I: 50 HCV negative subjects (age range 36-79), and Group II: 50 HCV positive subjects (age range 42-80). Expressions of the telomerase, retinoblastoma (Rb), E2F3, TP53 and FGFR3 genes were tested by immunohistochemistry and real time PCR in tumour tissues and healthy bladder tissues. Also, telomerase activity was assessed by telomeric repeats amplification protocol (TRAP).

Results: Bladder tumors associated with HCV infection were of high grade and invasive squamous cell carcinomas (SCCs). Expressions of hTERT, Rb, E2F3, TP53 and FGFR3 as well as telomerase activity were significantly higher in bladder tissues of HCV-infected patients compared with bladder tissues of non infected patients (p<0.05). On the contrary, CDKN1A (p21) expression was significantly lower in bladder tissues of HCV-infected patients compared to bladder tissues of non infected patients (p<0.05).

Conclusion: The expressions of hTERT, Rb, E2F3, TP53 and FGFR3 as well as the activity of telomerase were significantly high in malignant bladder tissues associated with HCV infection. On the other hand, CDKN1A (p21) expression was low in bladder tissues of HCV-infected subjects. Moreover, there was a positive correlation between HCV infection and expression of telomerase, E2F3, TP53 and FGFR3. There was a negative correlation between HCV infection and expression of Rb and p21.

Figure 1

Specimens of Transitional cell carcinoma (TCC). A = TCC with negativity for telomerase (hTERT), B = TCC with intranuclear dot like positivity of hTERT in more than 25% of tumour cells from non-HCV infected patients, and C = TCC with intranuclear dot like positivity in more than 50% of tumour cells from HCV infected patients (immunoperoxidase DAB X400).

Figure 2

Specimens of Bladder Cancer. A = normal Rb heterogenous nuclear staining in less than 50% of tumor cells invasive carcinoma from non-HCV infected patients, B = altered Rb expression; homogenous strong staining for Rb in more than 50% of tumor cells from HCV infected patients and C = BC with negativity for Rb from HCV- infected patients (immunoperoxidase DAB X400).

Figure 3

Specimens of bladder urothelium. A = non-neoplastic urothelium with nuclear immunoreactivity for p53 in more than 10% of urothelial cells, B = BC with nuclear immunoreactivity in less than 10% of tumor cells (scored negative) from non-HCV infected patients and C = BC with increased p53 expression in more than 10 of tumour cells in invasive carcinoma from HCV-infected patients (Immunoperoxidase DAB X 400).

Figure 4

Specimens of bladder urothelium. A = non-neoplastic urothelium with no nuclear expression for p21 from HCV-infected patients, B = Transitional cell carcinoma (TCC) with nuclear immunoreactivity for p21 in more than 10% of tumor cells from non-HCV infected patients and C = lost p21 expression in most tumor cells from HCV infected patients (Immunoperoxidase DAB X 400).

Figure 5

Specimens of BC. A = urothelial carcinoma with faint but detectable membranous and cytoplasmic positivity in tumor cells for FGFR3 from non-HCV infected patients, B = BC with moderate cytoplasmic positivity in most tumor cells for FGFR3 from HCV infected patients and C = BC with strong cytoplasmic positivity in tumor cells for FGFR3 from HCV infected patients (Immunoperoxidase DAB X400).

Figure 6

Kaplan-Meier analysis for cases of BC. Patients with HCV infections showed significantly lower overall survival rate (A), and disease free survival (positive case) (B).