Effects Of Proliferation And Migration In Hepatocellular Carcinoma Cells After Silencing Paip1 Expression

Objective:In the early stage, we used microarray technology to screen the expression of Paip1 protein in hepatocellular carcinoma(HCC) and adjacent liver tissues.This study we detect and observe the cell proliferation and migration of hepatocellular carcinoma cells when we silence Paip1 gene and the relative expression of Paip1 protein was downregulated.Methods:1. Western blots method was used to detect the expression of Paip1 protein in four HCC cell lines Hep G2, Hep G2.2.15, SMMC-7721, Hep3 B and human liver cell line L02.2. We silence Paip1 gene of HepG2 cells and SMMC-7721 cells by using lentivirus mediated RNA interference(RNAi),the efficiency of infection was observed by inverted fluorescence microscope and was tested by western blot method.In addition,we use puromycin screening stable lentivirus infection cells(SMMC-7721-LV-sh-Paip1 and Hep G2-LV-sh-Paip1, SMMC-7721-sh-e GFP and Hep G2-sh-e GFP),cell scratch healing experiment and CCK-8 assay to observe the change of migration area and proliferation ability of the cells when Paip1 gene silencing; Western blot was used to detect PDCD4 protein levels of cells when Paip1 gene silencing.Results:1.The expression of Paip1 protein in HCC cell lines Hep G2, Hep G2.2.15,SMMC-7721, Hep3 B are significantly higher than that in human liver cell line L02(P< 0.05), we selected two HCC lines(Hep G2 and SMMC-7721) to complete the next cell experiments.2.After LV-LV-sh-GFP infected Hep G2 cell(and SMMC-7721),the fluorescence microscope was used to observe the expression of GFP, and the 90% cells lighted green fluorescence.The LV--sh-Paip1 could effectively inhibited the expression of Paip1 protein levels.3.after 24 hours scratching, changes of the migration area of Hep G2 cells which infected LV-sh-Paip1 than the corresponding negative control group and blank control group are smaller(p <0.01),so as SMMC-7721 cells.4. The proliferation of HepG2 cells which infected LV-sh-Paip1 than the corresponding negative control group and blank control group are weaker(inhibition rate is 48.60%),so as SMMC-7721 cells(inhibition rate is 52.32%).5.In addition, The results of western blot show that PDCD4 protein relative expression levels of SMMC-7721 cells which infected LV-sh-Paip1, compared with the negative control group and blank control group are significantly higher(p <0.01).Conclusion:1.Paip1 protein levels of HCC cell lines Hep G2, Hep G2.2.15, SMMC-7721, Hep3 B are significantly higher than normal human liver cell line L02, indicating Paip1 may be highly expressing in HCC.2.RNAi interference technique was used to down regulate the expression of Paip1 gene, the migration and proliferation ability of Hep G2 cells and SMMC-7721 cells decrease significantly.3.By silencing Paip1 in SMMC-7721 cells, the expression of PDCD4 protein relative increase, while now PDCD4 is widely recognized as a disincentive in the development of HCC, preliminary verifying speculation that Paip1 may promote development of HCC in conjunction with PDCD4.