Effect Of ShRNA Targeting COX-2on The Expression Of Apoptosis And Lipometabolism Related Genes In HSC-T6Cells

ObjectiveResearch the effect of COX-2shRNA on the gene Ccl2, Phlda3, Aldoc andCar9which express in hepatic stellate cells, apoptosis or lipid metabolism, test andverify the results of the previous gene microarray.Methods1.There are three groups in these study: blank control group, empty vectortransfection group, COX-2shRNA1group.2. Use the constructed pYr-1.1-hU6-EGFP-COX-2shRNA1, and then transferit into HSCs, assess the transfection efficiency under the fluorescence microscope.3. Use PCR method to detect the expression of Ccl2, Phlda3, Aldoc and Car9after24h,48h,72h.4. Contrast the detected results with the previous gene microarray and thenanalyse every gene.Results1. The fluorescence microscope show that we can see green fluorescence inHSCs,cytoplasm after24h,48h,72h by COX-2shRNA1transfected into HSC.2. The expression of COX-2in different groups in HSC: the expression lever ofCOX-2mRNA in COX-2shRNA1group is lower than blank group and HK plasmidcontrol group after24h,48h,72h by COX-2shRNA1transfected (P<0.05), and thedecrease of COX-2shRNA1group is the most remarkable. 3. The expression of apoptosis genes in different groups in HSC: the expressionlever of Ccl2in COX-2shRNA1group is lower and Phlda3is higher than blank groupand HK plasmid control group after24h,48h,72h by COX-2shRNA1transfected (P<0.05), and the group of72h is the most prominent. There is no difference betweenblank group and HK plasmid control group.4. The expression of lipid metabolism genes in different groups in HSC: theexpression lever of Aldoc, Car9in COX-2shRNA1group is lower than blank groupand HK plasmid control group after24h,48h,72h by COX-2shRNA1transfected (P<0.05), and the group of72h is the most prominent. There is no difference betweenblank group and HK plasmid control group.5. From the previous gene microarray we can know that the expression ofPhlda3, Aldoc and Car9in this experiment is consistent with the gene microarray, butCcl2is contrary.Conclusions1. The COX-2shRNA can silence the expression of COX-2in HSC.2. The COX-2shRNA can promote the expression of apoptosis genes Phlda3inHSC.3. The COX-2shRNA can restrain the expression of lipid metabolism genesAldoc and Car9in HSC.4. The COX-2shRNA may regulate the expression of Phlda3, Aldoc, Car9topromote apoptosis of HSC.