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目的:探讨新型凋亡分子TNF相关凋亡诱导配体(TRAIL)对人肝癌细胞系的作用及生物学活性的影响。方法:免疫细胞化学法检测HepG2细胞表面膜结合型TRAIL的表达,ELISA法检测HepG2细胞分泌型TRAIL的表达。MTT和TUNEL法分别进行细胞毒实验和凋亡的检测。HepG2细胞内端粒酶的活性由TRAP-PCR法检测,端粒酶催化亚单位hTERT的表达由流式细胞术进行检测。结果:HepG2肝癌细胞系表面组成性表达TRAIL分子,并有适量的sTRAIL呈分泌表达。细胞毒实验显示TRAIL能够显著抑制肝癌细胞的生长,TUNEL检测结果显示TRAIL具有诱导肝癌细胞凋亡的效应。端粒酶的活性检测显示,TRAIL能够有效抑制肝癌细胞系内端粒酶的活性以及端粒酶催化亚单位的表达。结论:TRAIL是一个有效的抑癌分子,能够通过诱导细胞凋亡、抑制端粒酶活性等途径控制肿瘤细胞的生长和进一步杀伤肿瘤细胞。
Objective: To investigate the effect of TNF-related apoptosis-inducing ligand (TRAIL) on human hepatoma cell line and its biological activity. Methods: Immunocytochemistry was used to detect the expression of membrane bound TRAIL in HepG2 cells. The expression of secreted TRAIL in HepG2 cells was detected by ELISA. MTT and TUNEL assay cytotoxicity and apoptosis were detected. The activity of telomerase in HepG2 cells was detected by TRAP-PCR. The expression of telomerase catalytic subunit hTERT was detected by flow cytometry. Results: Hepatocellular carcinoma cell line HepG2 constitutively expressed TRAIL molecules and secreted sTRAIL. Cytotoxicity assay showed that TRAIL can significantly inhibit the growth of hepatoma cells, TUNEL assay showed that TRAIL has the effect of inducing apoptosis of hepatoma cells. Detection of telomerase activity showed that TRAIL can effectively inhibit the activity of telomerase and the expression of telomerase catalytic subunit in hepatoma cell line. Conclusion: TRAIL is a potent tumor suppressor molecule that can control the growth of tumor cells and kill tumor cells by inducing apoptosis and inhibiting telomerase activity.