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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。
Objective: To investigate the effect of short hairpin RNA (shRNAs) on the expression of human telomerase catalytic subunit (hTERT). Methods: The oligonucleotides encoding shRNA targeting hTERT were cloned into mammalian shRNA expression vector pUC18U6 to form pUC18U6ht, and then transfected into HepG2 cells by liposome. HepG2 cells transfected with pUC18U6 and pUC18U6GFPsir expressing anti-green fluorescent protein shRNA served as controls. The hTERT mRNA of transfected cells was quantitated by real-time quantitative RT-PCR. Results: shRNA reduced the hTERT mRNA levels by 49% (P <0.05) compared to the control. Conclusion: shRNAs inhibit hTERT expression.