Effect Of COX-2Small Hairpin RNA On The Chemokine Expression In HSC-T6Cell

Objective:To investigate the influence of COX-2small hairpin RNA on the chemokineexpression in HSC-T6cell.Methods:1、There are three groups in these study: blank control group, empty vectortransfection group, COX-2shRNA1group.2、The27K Rat Genome Array from Beijing CapitalBio Corporation to findingdifferent expressed chemokine genes in empty vector transfection and COX-2shRNA1group in different time.3、The extraction and concentration of total RNA in hepatic stellate cells wereutilized respectively by Trizol. Use PCR method to detect the different expressiongene from the gene microarray.Results:1、The fluorescence microscope show that we can see green fluorescence inHSC-T6cells, cytoplasm in different time by COX-2shRNA1transfected intoHSC-T6cells, and the transfection efficiency was above70%;2、According to the standard of difference in gene chip fluorescence ratio of1.5times (Ratio values≥1.5and≤0.667),the expression of CXCL11、CX3CL1、CXCL10and CCL2were increased at48hours by COX-2shRNA1transfected in genemicroarray in HSC-T6cells;3、According to the standard of difference in gene chip fluorescence ratio of1.5times (Ratio values≥1.5and≤0.667),the expression of CX3CL1and CXCL10wereincreased at72hours by COX-2shRNA1transfected in gene microarray in HSC-T6cells; 4、RT-PCR results showed the expression of CX3CL1was increased andCXCl10、CXCL11、CCL2were decreased after COX-2shRNA1transfected into theHSC-T6cells at72h (p <0.05).Conclusions:1、The gene expression profile of chemokines take place changes after silenceCOX-2by shRNA in HSC-T6cells;2、The expression of chemokine gene is not the same in HSC-T6after theCOX-2shRNA1transfected into HSC-T6cells;3、RT-PCR test showed CX3CL1mRNA expression was significantly increased,and CXCL10mRNA、CXCL11mRNA、CCL2mRNA were significantly decreasedafter COX-2shRNA1transfected into the HSC-T6cells at72h.