南方医科大学学报

南方医科大学学报 ›› 2014, Vol. 34 ›› Issue (07): 917-.

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曲古抑菌素对HepG2细胞增殖的影响及相关机制

石庆强，左国伟，冯子强，赵绿翠，罗念，游智梅，夏菁，李丹阳，李静，陈地龙   

  • 出版日期:2014-07-20 发布日期:2014-07-20

Inhibitory effect of trichostatin A on HepG2 cell proliferation and the mechanisms

  • Online:2014-07-20 Published:2014-07-20

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摘要： 目的研究曲古抑菌素（TSA）对HepG2 细胞凋亡的影响及可能的机制。方法取对数生长期HepG2 细胞，TSA（50~
500 nmol/L）诱导，同时设空白对照组，用CCK-8检测细胞增殖影响；流式细胞术检测细胞周期；Annexin V-FTIC/PI双染细胞检
测凋亡；倒置显微镜观察细胞形态并采图；Western blot检测HepG2细胞中β-catenin、HDAC1、HDAC3、H3K9、CyclinD1、Bax等
蛋白的表达。定量PCR方法检测HDAC1，HDAC3基因的表达。结果CCK-8检测结果显示TSA对肝癌HepG2细胞生长抑制
呈剂量和时间依赖性；流式细胞术检测细胞周期表明，药物组细胞G0/G1期所占比例增加，S期所占比例下降，G2/M所占比例增
加；细胞凋亡检测表明，空白对照组凋亡率（6.22±0.25）%；250 nmol/L 组凋亡率（7.17±0.20）%；500 nmol/L 组凋亡率（18.14±
0.42）%；倒置显微镜下观察可见，随着时间推移空白对照组细胞生长良好；而药物组细胞则出现变形，有大量漂浮细胞；Western
blot检测结果显示β-catenin、H3K9和Bax蛋白表达水平均明显升高，CyclinD1、HDAC1、HDAC3的表达水平则呈下降趋势；定
量PCR结果显示：HDAC1、HDAC3 基因的表达无明显变化。结论TSA可通过抑制HDAC的活性，促进组蛋白乙酰化，活化
Wnt/β-catenin信号通路来发挥其抑制HepG2细胞增殖，诱导周期阻滞及凋亡的作用。

Abstract: Objective To investigate the inhibitory effect of trichostatin A (TSA) on the proliferation of HepG2 cells and explore
the underlying mechanism. Methods HepG2 cells exposed to different concentrations of TSA for 24, 48, or 72 h were examined
for cell growth inhibition using a cell counting kit, changes in cell cycle distribution with flow cytometry, cell apoptosis with
annexin V-FTIC/PI double staining, and cell morphology changes under inverted microscope. The expressions of beta-catenin,
HDAC1, HDAC3, H3K9, cyclinD1 and Bax proteins in the exposed cells were detected by Western blotting, and the
expressions of HDAC1 and HDAC3 mRNAs by quantitative fluorescent PCR. Results Exposure to TSA caused significant
dose- and time-dependent inhibition of HepG2 cell proliferation (P<0.05) and resulted in increased cell percentage in G0/G1 and
G2/M phases and decreased cell percentage in S phase. The apoptotic index in the control group was (6.22 ± 0.25)% , which
increased to (7.17±0.20)% and (18.14±0.42)% after exposure to 250 and 500 nmol/L TSA, respectively. Exposure to 250 and 500
nmol/L TSA also caused cell morphology changes with numerous floating cells. The expressions of beta-catenin, H3K9 and Bax
proteins were significantly increased and CyclinD1, HDAC1, and HDAC3 protein expressions decreased in TSA-treated cells,
but the expressions of HDAC1 and HDAC3 mRNAs showed no significant changes. Conclusion TSA can inhibit the
proliferation of HepG2 cells and induce cell cycle arrest and apoptosis by inhibiting HDAC activity, promoting histone
acetylation, and activating Wnt/beta-catenin signaling pathway.