短发夹RNA对人端粒酶催化亚基表达的影响

The effect of short hairpin RNA on hTERT expression

目的:研究短发夹RNA(shRNAs)对人端粒酶催化亚基(hTERT)表达的影响。方法:将编码针对hTERT的shRNA的寡聚核苷酸克隆入哺乳细胞shRNA表达载体pUC18U6形成pUC18U6ht,然后以脂质体法转染入HepG2细胞。被pUC18U6和表达抗绿色荧光蛋白shRNA的pUC18U6GFPsir转染的HepG2细胞作为对照。转染细胞的hTERTmRNA以实时定量RT-PCR定量测定。结果:与对照相比,shRNA可使hTERTmRNA的水平降低49%(P<0.05)。结论:shRNAs抑制了hTERT的表达。

AIM: To study the effect of short hairpin RNA (shRNA) on hTERT expression. METHODS: Oligonucleotides encoding shRNA against hTERT was cloned into a mammalian shRNA expression vector pUC18U6 to form pUC18U6ht which was transfected into HepG2 cells by using liposome. HepG2 cells transfected by pUC18U6 and pUC18U6GFPsir which expressed shRNA against green fluorescent protein were used as controls. hTERT mRNA in the transfected cells was quantified by using real-time fluorescent RT-PCR. RESULTS: Compared with the controls, the short hairpin RNA against hTERT decreased the hTERT mRNA level by 49% (P<0.05). CONCLUSION: hTERT expression was reduced by the shRNA.