RNAi inhibit the expression of S100A4 gene and its effect on invasion and metastasis ability of osteosarcoma cell in vitro

[Objective]To observe its effect on proliferation and invasion capacity in vitro of MG-63 cells,by application of RNAi technology to inhibit the expression of S100A4 gene in human osteosarcoma MG-63 cells.[Methods]The siRNA vector of S100A4 was constructed.According to target mRNA sequence,three RNA interference and non-specific target sequence of siRNA were designed.The cultured cells were divided into Group C(control group),group C1(liposome transfection group),group C2(non-specific siRNA transfection group),groups S1,S2,S3(transfected with specific siRNA Ⅰ,Ⅱ,Ⅲ).The situation of transfection was observed by fluorescence inverted microscope.The changes of S100A4 gene expression of the cells before and after transfection were detected by real-time PCR method.The changes of S100A4 protein expression before and after transfection were detected by Western-Blot.The specific siRNA with high transfection efficiency was selected and used as the transfection experiment group for the following tests.The following test was divided into normal cell group,negative control group and experimental group.The changes of cell proliferation were determined by MTT and flat colony-forming experiment.The changes of cell invasion and metastasis ability were determined by transwell chamber assay.[Results]Green fluorescence could be seen in cytoplasm of group C2 and group S1,S2,S3 by fluorescence inverted microscope.The expression levels of S100A4 mRNA in each specific siRNA transfection group were down regulated at various degrees.The group S3 was the most significant one.The expression levels of S100A4 protein of Western-blot detection were similar to the conclusions.MTT method and flat colony-forming experiments had shown that cell proliferation was inhibited in group S3.There was significant difference compared with the normal cells group and non-specific group(P0.05).Transwell chamber experiments showed that the invasive ability of the cells significantly decreased after RNA interference S100A4 gene expression,the cell number through artificial basement membrane decreased significantly,compared with the normal cell group and negative group(P0.05).[Conclusion]S100A4 siRNA-specific vector in vitro synthesis can effectively inhibit the expression of S100A4 in osteosarcoma MG-63cells,and cell proliferation and invasive ability in vitro.