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目的 研究重组人肿瘤坏死因子 (rhTNF)对端粒酶活性的调节作用。方法 将不同浓度rhTNF加入肝癌细胞株HepG2和HepG1 6细胞 ,以TRAP ELISA方法测定其端粒酶活性 ;采用Lipofect脂质体转染法将携带端粒酶催化亚单位—端粒酶逆转录酶 (hTERT)启动子的质粒转染至肝癌细胞株HepG2细胞 ,2h后加入不同浓度的rhTNF ,孵育 48h后 ,分别检测其报告基因萤虫素酶活性。 结果 在 10～ 10 0 0IU/ml浓度范围内 ,rhTNF可抑制端粒酶活性 ,同时在该浓度范围内 ,rhTNF可显著抑制hTERT启动子的表达 ,其抑制作用与rhTNF剂量呈正相关。结论 rhTNF杀伤肿瘤细胞的机制可能与其抑制hTERT的表达有关。
Objective To study the regulatory effect of recombinant human tumor necrosis factor (rhTNF) on telomerase activity. Methods Different concentrations of rhTNF were added into HepG2 and HepG1 6 cell lines, and the telomerase activity was determined by TRAP ELISA. Lipofect liposome transfection method was used to carry telomerase catalytic subunit-telomerase reverse transcriptase hTERT) plasmid was transfected into HepG2 cells. After 2 h, different concentrations of rhTNF were added. After incubation for 48 h, the reporter gene activity of luciferase was detected. Results The concentration of 10 ~ 10 0 IU / ml rhTNF inhibited telomerase activity. At the same time, rhTNF inhibited the expression of hTERT promoter in a dose - dependent manner. The inhibitory effect was positively correlated with the dosage of rhTNF. Conclusion The mechanism of rhTNF killing tumor cells may be related to its inhibition of hTERT expression.