Rnai Silencing The Cd147 Gene Of The Human Laryngeal Cancer Cell Proliferation, Invasion And Cisplatin Sensitivity And Body-tumorigenic Effects

AIM:CD 147, also named extracelluar matrix metalloproteinase inducer (EMMPRIN), is a member of the immunoglobulin family and a glycoprotein enriched on the surface of tumor cells, which promotes invasion, metastasis, growth and survival of malignant cells, and is known to confer resistance to some chemotherapeutic drugs. To determine the possible role of CD 147 in the invasive properties of laryngeal carcinoma, we used an RNA interference approach to silence CD 147 expression in the Hep2 cell line at high levels of CD 147 expression.Methods:1) The recombinant plasmid pSilencer/shRNA-control,pSilencer/shRNAl pSilencer/shRNA2 were stably transfected into Hep2 cells, and the change of the expression of CD147 mRNA and protein in transfected cells was identified by RT-PCR and Wersternblot;2) The change of proliferation level of the Hep2 cells and chemosensitivity to anti-tumor drug cisplatin after transfection was detected by MTT assay;3) The effect of CD147 siRNA on invasion in Hep2 cells in vitro was detected by transwell chamber;4) The effect of CD 147 silencing on the enzyme activities of MMP-2 and MMP-9 was identified using gelatin zymography5) The effect of CD147siRNA on potential of invasion in Hep2 cells in vivo were observed using nude mice subcutaneous xenotransplantation model.Results:1) The Laryngeal carcinoma cell strain Hep2 was transfected by pSilencer/shRNA-control, pSilencer/shRNAl and pSilencer/shRNA2 respectively. After selected by G418, the positive clone was successfully got; The results of RT-PCR and western blot showed that CD 147 expression was significantly impeded at both mRNA and protein levels.2) MTT assay showed that the proliferation of Hep2/shRNAl and Hep2/shRNA2 was inhibited to 77.34%(P0.01)and 70.43%(P<0.01),62.99%(P<0.01) and 73.76%(P<0.01),67.49%(P<0.01) and 61.37%(P<0.01) at 72,96 and120h, respectively. Also the chemosensitivity to cisplatin in Hep2/shRNAl and Hep2/shRNA2 was markedly increased compared with Hep2. 3) In vitro invasion assay showed that invasion cell number in Hep2/shRNAl groop and Hep2/shRNA2 groop was obviously lower than Hep2 groop and Hep2/shRNA-control groop.4) The gelatinolytic activities of both MMP-2 and MMP-9 were found to be reduced in Hep2/shRNAl and Hep2/shRNA2 (P<0.01).5) In mice inoculated with Hep2/shRNAl and Hep2/shRNA2, the tumor volume was 38.35%and 36.08%of that recorded for mice transplanted with Hep2 cells (P<0.01), respectively.Conclusion:1) After CD 147 experssion of Hep2 cells were suppressed, the cells proliferation ability was inhibited significantly, which was indicated that CD 147 could promote the growth of laryngeal carcinoma cells.2) CD 147 expression sensitized cells to cisplatin, this result indicated that CD 147 was a laryngeal carcinoma-related gene and CD 147 might be a potential target for therapeutic anti-cancer drugs.3) After CD 147 experssion of Hep2 cells were suppressed, the cells invasion ability and tumorigenesis in nude mice were both inhibited significantly, which indicated that CD 147 is associated with the progression of laryngeal carcinomas and might be a new potential target for therapeutic it.