The Effects And Mechanisms Of VTCN1Gene In Bladder Cancer

Objective:Some tumor cell lines also showed constitutive expression VTCN1. In lung cancer, ovarian cancer, renal cell carcinoma were VTCN1protein; breast and endometrial cancer VTCN1almost ubiquitous expression does not depend on the tumor grade and stage, and VTCN1high expression of gastric cancer is significantly increased risk of death independent factor. But VTCN1in bladder tumor tissue in the exact clinical significance remains to be further verified, VTCN1whether as a bladder cancer early diagnosis and prognostic evaluation indicators also deserves further study. VTCN1negative molecule gene family in tumor suggesting that abnormal expression in tumor development plays an important role in the mechanism of which will be involved in tumor development interventions provide an effective way, with great potential value. A growing number of clinical studies have found VTCN1molecules are closely related with the regulation of tumor immunity, but tumor immune response in the body in the mechanism of action remains to be further studied. Therefore, Part1was to investigate the relationship of the expression of VTCN1in bladder cancer tissues and its clinical pathological characteristics and the prognosis of bladder cancer patients. Part2was to study the effect of inhibitor of apoptosis VTCN1down-regulation by small interfering RNA-mediated RNA interference (RNAi) on the biological features of bladder carcinoma cell line HT-1376.Methods:Part1:The content of sVTCNl protein was detected in the serum of patients with bladder cancer by enzyme immunoassay (ELISA), and the expression of VTCN1mRNA was detected by real time quantitative PCR, and the VTCN1expression levels in the tissue samples of patients with bladder cancer was detected by immunohistoche-mical staining.Part2:HT-1376cells were transfected with synthetic small interfering RNA (siRNA) targeting VTCN1. Expression of VTCN1mRNA and protein were respectively measured by reverse transcription polymerase chain reaction (RT-PCR) and Western blot, Transfection efficiencies were monitored by those ways. The distribution of cell cycle phases was determined using flow cytometry. The proliferative and invasive ability of HT-1376cells in vitro was evaluated by the colony-forming unit assay and Transwell migration assay respectively.Results:Part1:The content of VTCN1protein (45.25±8.93μg/L) in the serun of patients with bladder cancer was significantly higher than the patients with bladder benign tumors (34.41±6.91μg/L) and the normal population (31.20±6.28±g/L)(P <0.01). VTCN1mRNA expression in bladder cancer tissue was significantly higher than the adjacent tissues of bladder cancer (P<0.01), and the expression of VTCN1mRNA in the adjacent tissues of bladder cancer was significantly higher than the bladder benign tumors (P<0.01). Further to detect the VTCN1protein expression in70cases of bladder cancer tissues, the positive rate was68.57%, including77.08%patients were highly expressed. The positive rate in the patients with bladder cancer was significantly higher than the bladder benign tissue (P<0.01). The expression of VTCN1protein were significance with TNM classification and distant metastasis of bladder cancer(P<0.05), but no significance with gender, age, histological type, clinical stage (P>0.05). The survival rate with highly expression VTCN1in patients was lower than with low-expression patients (P<0.05).Part2:Both VTCN1mRNA and protein expression were significantly decreased in the experimental group compared with controls (P<0.05). The cell cycle was arrested in the Gl phase. The proliferation and invasiveness of treated HT-1376cells were inhibited in vitro.Conclusions:(1) The expression of VTCN1in bladder cancer was correlated with the metastasis and prognosis of bladder cancer, and VTCN1could be used as an effective indicator to reflect the biological behavior of bladder cancer and prognosis.(2) Delivery of siRNA targeting VTCN1seems efficient in down-regulating VTCN1expression and diminishing the growth, proliferation and invasiveness of HT-1376cells, suggesting that siRNA-based strategy targeting VTCN1may play a foundation for the clinical management of bladder carcinoma.