TMPyP4对肝癌细胞HepG2生长抑制作用的研究  

Inhibitory effect of TMPyP4 on HepG2 cell growth

作　　者：阎敬[1] 贺安宁[2] 郝凤进[1] 关一夫[1] 

机构地区：[1]中国医科大学基础医学院生物化学与分子生物学教研室,辽宁沈阳110122 [2]中国医科大学附属第一医院肿瘤研究所第一研究室,辽宁沈阳110001

出　　处：《中华肿瘤防治杂志》2016年第12期775-779,共5页Chinese Journal of Cancer Prevention and Treatment

基　　金：国家自然科学基金(31070705);辽宁省科技厅项目(2013225089)

摘　　要：目的 TMPyP4是G-四链体的配体,能够诱导形成G-四链体结构并增强G-四链体结构的稳定性,加强G-四链体作为负调控因子抑制MET基因的表达;G-四链体结构的形成可以选择性的抑制肿瘤细胞增殖。本研究旨在分析TMPyP4对肝癌细胞HepG2生长抑制作用的研究,为MET高表达的肿瘤靶向治疗提供新思路。方法采用CCK-8方法分析HepG2细胞增殖变化,通过Hoechst 33342细胞核染色和AnnexinⅤ-FITC/PI双染流式细胞术分析HepG2细胞凋亡水平,以及利用流式细胞术分析HepG2细胞周期分布的变化。结果 CCK-8分析HepG2细胞增殖显示,与对照组比较,10μmol/L TMPyP4对HepG2细胞存活率无显著影响,20、50、100和200μmol/L TMPyP4组HepG2细胞存活率分别为(82.28±1.99)%、(68.88±3.06)%、(55.57±2.76)%和(50.32±5.78)%,F=120.542,P<0.001,说明高于20μmol/L的TMPyP4显著抑制细胞增殖;流式细胞术分析细胞凋亡不明显,只有高浓度TMPyP4诱导少量HepG2细胞发生了凋亡,并出现了坏死细胞;100和200μmol/L TMPyP4的实验组细胞经Hoechst 33342染色,有少数HepG2细胞表现出细胞核固缩、致密,形态不规则,染色不均一,呈现凋亡表现;不同浓度的TMPyP4处理HepG2细胞使G0/G1期细胞增加了20%,S期细胞减少,G2/M期细胞也减少,表示HepG2细胞发生了G0/G1细胞周期阻滞。结论 G-四链体的配体TMPyP4显著抑制了肝癌细胞HepG2的细胞增殖,诱导少量HepG2细胞凋亡,并使HepG2细胞发生了G0/G1细胞周期阻滞。OBJECTIVE TMPyP4 is a ligand of G quadruplex, which can stabilize the G-quadruplex structure in the promoter of MET gene and enhance its inhibitory effect on its expression. The G-quadruplex structures of protooncogenes can inhibit the proliferation of cancer cell. This study aims to analyze the effect of TMPyP4 on HepG2 cell growth by sta bilizing the G-quadruplex structure in MET promoter, which will provide a new way for targeting tumor therapy. METH- ODS CCK-8 assay was performed to analyze the proliferation of HepG2 cell. Hoeehst 33342 dyeing for nucleus was used to reveal the cell apoptosis. The flow cytometry was utilized to show the cell apoptosis and the cell cycle distribution. RESULTS CCK-8 assay showed that the cell viabilities in 20, 50, 100 and 200 μmol/L TMPyP4 groups were （82.28± 1.99） %, （68.88 ±3.06）% , （55.57±2.76）% and （50.32± 5.78）%, indicating the significant inhibitory effect on cell proliferation （F= 120. 542, P〈0. 001）. But 10 μmol/L TMPyP4 had no effect on the cell proliferation of HepG2. Minor cell apoptosis only happened when being treated by high concentration of TMPyP4, in which nucleus exhibited chromatin pycnosis, uneven dyeing by Hoechst 33342 and irregular shape. HepG2 cells at G0/G1 phase increased by 20% and decreased at G2 and S phase, which indicated that TMPyP4 induced the cell cycle arrest at G0/G1 phase. CONCLUSIONS TMPyP4 can stabilize G-quadruplex structure in the promoter of MET, significantly inhibit the HepG2 cell proliferation, induce G0/G1-phase cell cycle arrest and make minor HepG2 cell apoptosis.

关 键 词：TMPyP4 细胞增殖 细胞凋亡 细胞周期阻滞 MET启动子G-四链体 

分 类 号：R285[医药卫生—中药学]