Abstract
Objective To construct SiRNA recombinant expression vector targeting Livin gene and research the effect of it in inhibition of human colon cancer cells proliferation and enhancement of chemotherapy sensitivity.Methods SiRNA recombinant expression vector targeting Livin gone Was constructed and transfected into human colon cancer cell.The effect of SiRNA recombinant expression vector was detected by RT-PCR and Western blot.Using auto-enzyme-labeled reading device with the 490nm wave,the value of A in each foramen was respectively measured after 24,48,72 hr of transfection by MTT reduction assay,and then calculated the inhibitor rate of tumor cell.After 48 hours of transfection,the change of the apoptosis rate was detected under the 488nm wave by flow cytometry.Results It was confirmed by restriction endonuclease and sequence analysis that SiRNA recombinant expression vector targeting Livin gene was constructed successfully.Inhibition ratio of Livin SiRNA at mRNA and protein levels were 30.18% and 28.88%.It could enhance chemotherapy sensitivity by combining 5-Fu.Conclusion The SiRNA recombinant expression vector targeting Livin gene has been constructed successfully and can inhibite the expression of Livin gene.It also can enhance chemotherapy sensitivity and apoptosis induction effect by combining 5-Fu in human colon cancer cells significantly. Key words: Livin; RNA interference; Gene therapy; Chemotherapy
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