Lentivirus-mediated RNA Interfere Targeting RRS1 In Cholangiocarcinoma Cell Proliferation In Vitro

Objective:This study aimed at investigating whether lentivirus-mediated RNA has effect on cholangiocarcinoma cell proliferation af ter applying RNA interference technology to inhibit RRS expression in bile duct cancer RBE.Materials and Methods:1.Apply Real-time PCR method to dete ct RRS1 mRNA expression in 10 cholangiocarcinoma tissues and pr ecancerous tissues collected from patients in the people’s hospital of Hunan Province during the time from May to December in 2013. Ascertain whether RRS in cholangiocarcinoma have abnormal expr ession.2.Objective to investigate the effects of RRS1 expression inter ference on cholangiocarcinoma cell proliferation observed the effects of RRS1 interference on cholangiocarcinoma RBE cell proliferation in vitro.To detect cholangiocarcinoma mRNA expression after RRS 1 inhibition used the lentivirus-mediated RNA interference technolog y to silent RRS1 gene expression at transcriptional level and Real-ti me fluorescence quantitative polymerase chain reaction technique.Results:1.We found that in 10 cases of patients with cholang iocarcinoma tissues and paracancerous tissue, the expression of RR S1 mRNA in cancer tissues compared to the precancerous tissues w as significantly higher, and the difference has statistically significant (P<0.05)Suggesting RRS in cholangiocarcinoma are up-rugulated.2.Successfully interrupt RRS1 gene expression at transcriptional level by the RRS1 lentivirus-mediated RNA interference technology. Detect the expression of RRS1 mRNAn in experimental group(transf ected RRS1-shRNA)and control group(transfected shRNA lentivirus vector)by Real-Time PCR. The result showed that mRNA relative expression in experimental group (repressed RRS1) was obviously restrained, the difference has statistical significant (P<0.05). Estimat e the knockdown efficiency was 80.10%, suggesting that RNA distu rbed RRS1 successfully.3.In vitro,Check cells growth counting in control group and negative control group by Cellomics, draw the growth curve for eac h group, The results showed that the bile duct cancer cell proliferat ion activity was obvious decreased after RNA targeted inhibited th e expression of RRS1 in RBE cells.Conclusion1. To detect clinical samples with Real-Time Fluorescent PCR (RTF PCR). RRS1 mRNA expression in cholangiocarcinoma tissue i s obviously higher than the paracancerous tissues, which suggested t hat RRS1 was higher expressed in cholangiocarcinoma tissue.2. After Lentivirus RNAi interfered RRS1 expression in RBE c ells the proliferation in bile duct cancer significantly decreased. s uggesting RRS1 may promote the cholangiocarcinoma proliferation a bility.