Abstract
Survivin gene may be a good target for cancer gene therapy because it is over expressed in a variety of human tumors including human hepatocellular carcinoma but not in differen- tiated adult tissues. To explore the effects of the siRNA of survivin gene inducing apoptosis in human hepatocellular cancer cells, three siRNAs cpusiRNA1, cpusiRNA2 and cpusiRNA3 were designed and transferred into human hepatocellular carcinoma cell line HepG2 (HepG2) by lipofection. MTT test showed that the growth of HepG2 decreased when it was transfected with 25nM, 50nM, 100nM, 150nM, 200nM, 400nM siRNA respectively after 48 hours. And the change of mRNA and protein of survivin gene and p53 gene had been detected by RT-PCR and Western blot. Cells presented an increase in apoptosis index was assayed by flow cytometry. Small interfering RNA can exert a knockdown of survivin gene expression and up regulation of p53 gene to induce apoptosis and to inhibit the growth of HepG2.
Highlights
Hepatocellular carcinoma (HCC) was the most pernicious cancer with a high mortality rate
Many studies have shown that the survivin gene was a new member of inhibitors of the apoptosis protein (IAP) family, which had been implicated in both control of cell division and inhibition of apoptosis
Its blocking action on gene expression had been successfully observed in rat and human cells cultured in vitro, and the knockdown of genes in cells had been achieved [11,15].A study[16] had shown that 21-25 nt small interference RNA could mediate specific gene silencing in mammal cells
Summary
Hepatocellular carcinoma (HCC) was the most pernicious cancer with a high mortality rate. Several preclinical studies had demonstrated that downregulation of survivin expression, accomplished through the use of small interfering RNA to increase the apoptotic rate and to reduce tumor growth potential. It could and efficiently degrade mRNA, resulting in post uctranscriptional gene silencing (PTGS) [7,8,9,10,11,12,13,14, 15], which was a natural mechanism in organisms underlying the resistance to virus invasion and inhibition of transposon mobility. Three small interfering RNAs were designed according to the sequence of survivin gene and they were transferred into human hepatocellular carcinoma cell line HepG2 by lipofection to observe survivin and p53 gene expression changes and their effects on cell apoptosis and growth, Published Online February 2009 in SciRes. http://www.scirp.org/journal/jbise
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