Experiment Study Of Effect Of Antisense CD147 On Invasion Of Human Cholangiocarcinoma Cell Line

Objective:To investigate the effect of antisense CD147 on invasion of human cholangiocarcinoma cell line(QBC939).Materials and Methods:We reconstruct a eukaryotic expression vector containing the CD 147 cDNA in an antisense orientation. Human cholangiocarcinoma cell were divided into either a antisense CD 147 treated group, empty vector group, or control group. QBC939 in antisense CD147 treated group underwent transfect with asCD147-pcDNA3.1(-); QBC939 in empty vector group underwent transfect with pcDNA3.1(-); Control group treated by DMEM. Both of them were tested by MTT method for cell Proliferation. Real-time PCR was performed to detect both CD147mRNA, MMP-2mRNA, MMP-9mRNA and TIMP-2mRNA expression. Western-blotting was performed to detect both CD 147 protein, MMP-2 protein, MMP-9 protein and TIMP-2 protein expression.Results:We reconstructed a eukaryotic expression vector containing the CD 147 cDNA in an antisense orientation. MTT was used to determinate the cell growth rate and no reduction in the growth rate was found after infection with antisense CD 147 treated group (P>0.05). The method of real-time PCR:The expression of mRNA of CD 147 molecule was effectively inhibited in the antisense CD 147 treated group compared with empty vector group and control group (P<0.05), empty vector group compared with control group was no effect influence (P>0.05), The expression of mRNA of MMP-2 molecule was effectively inhibited in the antisense CD 147 treated group compared with empty vector group and control group (P<0.05), empty vector group compared with control group was no effect influence (P>0.05), The expression of mRNA of MMP-9 molecule was no effect influence in the antisense CD 147 treated group compared with empty vector group and control group (P>0.05), The expression of mRNA of TIMP-2 molecule was no effect influence in the antisense CD 147 treated group compared with empty vector group and control group (P>0.05). The method of Western-blotting:The expression of protein of CD 147 molecule was effectively inhibited in the antisense CD 147 treated group compared with empty vector group and control group (P<0.05), empty vector group compared with control group was no effect influence (P>0.05), The expression of protein of MMP-2 molecule was effectively inhibited in the antisense CD 147 treated group compared with empty vector group and control group (P<0.05), empty vector group compared with control group was no effect influence (P>0.05), The expression of protein of MMP-9 molecule was no effect influence in the antisense CD147 treated group compared with empty vector group and control group (P>0.05), The expression of protein of TIMP-2 molecule was no effect influence in the antisense CD 147 treated group compared with empty vector group and control group (P>0.05).Conclusion:1.Antisense CD 147 have no effect on proliferation of QBC939 cell.2. Antisense CD147 no effect on the expression of mRNA and protein of MMP-9 and TIMP-2 molecule in QBC939 cell.3. Antisense CD 147 can inhibited the expression of mRNA and protein of MMP-9 and TIMP-2 molecule in QBC939 cell, may be can influence its Cell invasive.