瘦素对HepG2细胞端粒酶及端粒酶反转录酶的调控作用研究  

The biological actions of leptin-induced activity of telomerase and telomerase reverse transcriptase in HepG2

作　　者：曹坤[1] 李海洋[2] 

机构地区：[1]贵州省肿瘤医院,贵州贵阳550002 [2]贵州医科大学附属医院,贵州贵阳550004

出　　处：《贵州医药》2016年第12期1242-1245,共4页Guizhou Medical Journal

基　　金：贵州省科学技术基金资助项目(黔科合J字[2011]2235号)

摘　　要：目的探讨瘦素(leptin)对人肝癌细胞株HepG2的增殖机制及对端粒酶(telomerase,TA)与端粒酶反转录酶(hTERT)活性表达的影响。方法噻唑蓝(MTT)比色法检测不同浓度leptin作用不同时间对HepG2细胞的增殖影响;流式细胞仪分析其细胞周期;TRAP-PCR银染法检测不同浓度leptin作用HepG2细胞后端粒酶的活性;实时荧光定量PCR、蛋白免疫印迹(Western-blot)分析leptin作用24h后hTERT mRNA及蛋白水平的表达情况。结果leptin可以促进HepG2细胞增殖,具有浓度-时间依赖性;流式细胞仪结果显示leptin可以改变HepG2的细胞周期,使S期比例升高;leptin可以活化HepG2细胞端粒酶,上调HepG2hTERT mRNA及蛋白水平的表达。结论leptin可能通过上调HepG2hTERT表达,增强端粒酶活性,改变肝癌细胞株HepG2的生物学行为。Objective To analyze the biological actions of leptin-induced activity of telomerase in HepG2 liver cancer cells and provide a new explanation for obesity-related HCC.Methods Effects of different concentrations of leptin（40ng/mL,80ng/mL,120ng/mL）on HepG2 were detected with colorimetric assay by Methyl thiazol tetrazoliu（MTT）after incubation periods of 24 h,48h,and 72 h.Flow cytometry was performed to assess cell cycle progression of different concentrations of leptin as stated above after each 24 hincubation period.Telomerase activity after different concentrations of intervention in HepG2 was assessed using TRAP-silver staining Telomerase Detection Kit.mRNA and protein expression level of hTERT after different concentrations of intervention in HepG2 were assessed using real-time RT-PCR and western blot analysis.Results Leptin could improve the proliferation rate of HepG2 cells.The effect was in does and time-depended partly by MTT method.In the flow cytometry analysis,leptin could promote HepG2 cells entering the S phase from the G0/G1 phase.It was found that leptin activated telomerase in a dose-dependent manner.leptin upregulated the expression of Human Reverse Tanscriptase（hTERT）at mRNA and protein levels.Conclusion Leptin could promote the proliferation of HepG2 cells and promote HepG2 cells entering the S phase from the G0/G1 phase.Leptin is a key regulator of the malignant properties of hepatocellular carcinoma cells through modulation of hTERT,a critical player of oncogenesis.

关 键 词：瘦素 HEPG2细胞 端粒酶 端粒酶反转录酶 

分 类 号：R977.9[医药卫生—药品]