CD147 Gene Silencing Influences Tumor Cell Proliferation And Invasion In Thyroid Papillary Carcinoma Cell Line K1

Objective:In order to filtered out the effective CD147-siRNA sequence which can inhibit effectively the expression of CD147 in human thyroid papillary carcinoma cell line K1. In following, we also investigated the effect of proliferation and invasion activity in human thyroid papillary carcinoma cell line K1 after the CD147 gene silencing.Thus may provide a novel target in gene therapy.Methods:To designed and synthesized three different pairs sequence of CD147-siRNA which were transfected into K1 cells to knockdown the expression of CD147 named experimental group S1,S2,S3,in addition including normal group and negative control group. mRNA and protein levels of CD147 were detected by RT-PCR and ELISA. And also detected the mRNA and protein levels of MMP7 and VEGF-C by RT-PCR and Western-Blot. At last,we investigate the effect of proliferation and invasion activity by Transwell chambers and MTT experiments after the CD147 gene silencing in human thyroid papillary carcinoma cell line K1.Results:Compared with group of normal and control, mRNA and protein level of CD147 in S1 group have no significant difference(P>0.05).In contrast, mRNA and protein level of CD147 dscrease significantly(P<0.05) in the group of S2 and S3,in which mRNA level decreases respectively 67.81%,72.48%;and protein level decreases respectively 31.65%,35.47%.Meanwhile,compared with group of normal and control, mRNA and protein level of MMP7 in group of S2 and S3 also dscrease significantly,in which mRNA level decreases respectively 50.25%,53.40%, protein level decreases respectively 41.58%,40.49%. Also mRNA and protein level of VEGF-C in group of S2 and S3 dscrease significantly,in which mRNA level decreases respectively 66.41%,56.57%;protein level decreases respectively 51.88%,41.57%.In MTT experiment,after CD147 gene silencing 48h,the inhibition rates of cell growth were 41.59%,40.86%;while after transfection 72h,the inhibition rates respectively were 39.39%,38.89%. In transwell chamber assay,after transfection 72h,cells of S2,S3 group throughing membrane were lower than S1 group,invasion inhibition rates were 35.87%,30.16%.but at the same time,there are no significant difference(P>0.05) among S1,normal and control group in the experiment of MTT and Transwell mentioned above.Conclusion:Construct three different pairs of CD147-siRNA,finding out that CD147-siRNA of S2 and S3 group sequence can silence effectively the expression of CD147 gene. And in further research,we found that CD147 gene silencing can decrease the expression of protein and mRNA in the gene of MMP7 and VEGF-C ,inhibiting the ability of proliferation and invasion in K1 cells .Therefore,we speculate that the RNAi technology against CD147 is expected to be a new gene therapy in papillary thyroid carcinoma.