The Effect of hmSD Knockdown on Proliferation and Migration of PC-3 Cell in Vitro
|Course||Pathology and Pathophysiology|
|Keywords||hmSD RNA interference Matrix metalloproteinase Prostate cancer|
Objective: To construct to express the plasmid of pGCsi-hmSD , transfected prostate cancer PC-3 cells , observe the silence hmSD gene PC-3 cell proliferation , migration and MMP-2 mRNA expression in gene therapy for prostate cancer targeting reference . Methods : RT-PCR was used to detect the hmSD mRNA expression in PC-3 cell lines ; plasmid construct expressing pGCsi-hmSD recombinant technology , the application of the eukaryotic cell transfection technology prostate cancer PC-3 cells transfected detect the transfection efficiency ; PCs the 3 cells after transfection pGCsi-hmSD plasmid , RT-PCR detection hmSD mRNA expression changes inverted changes in cell morphology was observed under a microscope , MTT detect its influence on cell proliferation , the change of flow cytometry apoptosis ; program change marks experimental and Transwell assay cell migration , RT-PCR detection the transfection pGCsi-hmSD plasmid PC-3 cells MMP-2 mRNA expression changes . Results: 1. Determined by restriction enzyme digestion and DNA sequence confirmed successfully constructed pGCsi-hmSD plasmid ; 2.PC -3 cells transfected with efficiency greater than 50% in each group , and no difference in each group transfection efficiency . Flow cytometry showed that 3. PC-3 cells transfected pGCsi-hmSD plasmid , RT-PCR detect hmSDmRNA expression levels to reduce the number of cells were observed under inverted microscope reduce shrinkage , MTT experimental results show that the cell proliferation inhibition promote apoptosis . The results of the scratch test and Transwell experiments PC-3 cells transfected with plasmid after cell migration pGCsi-hmSD reduce the RT-PCR results showed that MMP-2 mRNA expression was decreased . Conclusion: 1 . Successfully constructed a pGCsi-hmSD plasmid , the specific silencing the expression of PC-3 cells hmSD . 2.hmSD can promote the proliferation of PC-3 cells , inhibition of tumor cell apoptosis . 3. HmSD-RNAi can reduce the migration of PC-3 cells , may be associated with the inhibition of MMP-2 mRNA expression .