| Objective:Inhibit the expression of survivn gene in hepatocellular carcinoma cell line HepG2, to observe RNA interference targeting on the cell cycle and apoptosis of liver cancer cell and to study the significance of clinical care, offer probability on treatment of hepatoma.Methods:To structure survivin-siRNA aimed at human hepatocellular by chemosynthesis.Then si RNA was introduced into hepatocellular carcinoma cell line. Afterwards to study below.(1)Took the HepG2 cells into 6-well culture dish, and then divided them into 3 groups:low dose, medium dose and high dose, blending gently.Besides dilute Negative control expression vector pshRNA-survivin-NC into 3.2ug to use 200ulOPTI-MEM I substrate in negative control.(2)Took the HepG2 cells in 96-well culture dish, and then divided them into 4 groups:interference group, siRNA interference control group, null group, non-transfected group as comparation.To observe effection on each group.Collect cells after 48 h, measure growth case by MTT.detect cell apoptosis rate by flow cytometry,and analyse effection of siRNA on Hepatocarcinoma by flow cytometry.Results:Expression on non-transfected group and interference group wasn't inhibited compare to non-transfected group,no difference was found.While expression on surviving in interference group was inhibited obviously compared to interference control group.The jamming efficiency is more than 85%, which can illustrate Construction of vector was effective.It can inhibit the expression of survivin on transfection cell effectively. The effection on surviving gene silencing about HepG2 cell proliferation in the result of MTT shows that absorbance A of HepG2 cell reduced compare to the comparation group at the same time. Absorbance of HepG2 cell about 24 and 48 h interference group was under comparation group obviously. P<0.05.Conclusion:siRNA Transfection of HepG2 cell could be inhibited expression of survivin gene, lead liver cancer cells apoptosis, inhibited cell proliferation. |
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