The Expression Of CD147and MMP-9in Rat Pituitary Tumor Cells By RNAi Targeting PTTG

BackgroundPituitary tumor transforming gene,also known as an oncogene,was discovered in rat pituitary tumor cells by Pel.et al in1997. In the further research,we found that PTTG was highly expressed in various tumors including colon, ovary, testis, lung, and breast.PTTG played an important role in inhibiting separation of sister chromatid and the repairing of DNA. An overexpression of PTTG also induced the tumor angiogenesis,aggressive and metastatic.CD147, known as Extracellular matrix metalloproteinases inducer(EMMPRIN), is a member of the immunoglobulin super family.CD147is a transmembrane glycoprotein with two immunoglobulin-like domains and involves in various important signal transduction of cells.In the study of tumor aggressive and metastatic,we found that highly expression of CD147has an intimate relationships with tumor progression.CD147promote the tumor cells secretion matrix metalloproteinases,and enhance the aggressive and metastatic of tumor cells.Matrix Metalloproteinases-9(MMP-9) is a member of matrix metalloproteinases which plays an important role in tissue destruction and inflammation through degradation of matrix proteins and proteolytic activation of cytokines.The expression of MMP-9in tumor was more than the nomal tissual,at the same time,MMP-9participated in the process of tumor aggressive.PTTG,CD147and MMP-9all played important roles in tumorigenesis,the experiment through RNA interference targeting PTTG of rat pituitary tumor cells,to detect the expressions of CD147and MMP-9which were correlationed with the tumor aggressive.To research the relationships of PTTG,CD147and MMP-9in signal transduction of tumor cells.For further studying, the invasion and apoptosis of pituitary tumor cells by shRNA targeting PTTG. It had more clear definition about the function of PTTG in tumorigenesis.The experiment divided into two parts.I The expression of CD147and MMP-9in rat pituitary tumor cells by RNAi targeting PTTGObject To study the influences of expressions of CD147and MMP-9in GH3rat pituitary tumor cells by RNAi targeting PTTG Methods GH3cells were cultured with conventional method. Transfected was done with lentivirus according to the process of instructions. The GH3cells was divided into three groups,blank groups,nagtive groups,interference groups.At72h after transfection, the protein and mRNA of the GH3cells were extracted, Western blotting detect the expressions of PTTG protein,CD147protein,MMP-9protein,reverse transcription-polymerase chain reaction(PT-PCR) assay was applied to detect the expressions of PTTG mRNA,CD147mRNA,MMP-9mRNA. Results the protein and mRNA expressions of PTTG,CD147,MMP-9of the interference groups cells were less than the expressions of other groups cells (p<0.05), The expression levels of CD147and MMP-9were correlationed with the expression of PTTG (p<0.05), The expression levels ofMMP-9was correlationed with the expression of CD147(p<0.05). Conclusion RNAi targeting PTTG can inhibits the expressions of CD147and MMP-9of rat pituitary tumor cells. Ⅱ RNAi targeting PTTG induce invasion and apoptosis of rat pituitary tumor cellsObject To study the influence of the invasion and apoptosis of rat pituitary tumor cells by shRNA targeting PTTG. Methods GH3cells were cultured with conventional method. Transfection was done with lentivirus according to the process of instructions. The GH3cells were divided into three groups,blank group, nagtive group,interference group. At48h and72h after transfection.,transwell assay to detect tumor cells invasion of three groups, Terminal deoxynucleotidyltransferase-mediated nick end labeling (TUNEL) assay to detect the tumor cells apoptosis of the three groups. Results At72h after shRNA transfected the GH3cells, tumor cells invasion of interference groups were inhibited obviously more than the other group cells (p<0.05), and enhanced the apoptosis of the tumor cells of interference group more obviously than the other groups cells.Conclusion RNAi targeting PTTG can inhibit the invasion of rat pituitary tumor cells obviously.,at the same time,inhenced the apoptosis of tumor cells.