Inhibition of hTERT gene by siRNA in SGC7901 gastric-carcinoma cells lines

Background and purpose:Stomach cancer is one of the most common malignancy of human being.In our country,it is one of leading cancer death,5-yr survival rate for the patients is just 15%-20%.At present,we do not have effective first line prevent measure for the disease,it becomes more and more important to pursue a effective method to prevent and treat gastric cancer.In our in vitro study,the inhibition of gastric-carcinoma cell SGC7901 proliferation by RNA interference could be observed.Methods:The stable screening-inhibition technique of RNAi was adopted to stably inhibit the expression of hTERT.Small hairpin interfering RNA(shRNA) targeting hTERT gene was designed,recombinant plasmid pGenesil-shRNA-hTERT was constructed,and the plasmid was transfected into gastric-carcinoma SGC7901 cells that clones were selected by G418 in order to establish gastric-carcinoma cell lines with stable expression of pGenesil-shRNA-hTERT.Real-time RT-PCR,MTT and PCR-TRAP were utilized to detect the alterations of hTERT mRNA expressions,telomerase activity and cell proliferation.Results:The effectiveness of RNAi could be observed in gastric-carcinoma SGC7901 cells with stable expression of pGenesil-shRNA-hTERT,in these cell lines,the expression of hTERT mRNA was obviously down-regulated.Meanwhile,the telomerase activity was significantly decreased.gastric-carcinoma SGC7901 cell proliferation was significantly inhibited in pGenesil-shRNA-hTERT group compared to negative control group.Conclusions:RNAi may continually and stably suppress hTERT mRNA expression and cell proliferation,which is a potential new approach for gene therapy of neoplasm.