土槿皮乙酸阻滞HepG2肝癌细胞于G2/M期并抑制其侵袭和迁移  

Pseudolaric acid B induces G2/M arrest and inhibits invasion and migration in HepG2 hepatoma cells

作　　者：李帅 郭莲怡 LI Shuai1 , GUO Lianyi2(1Graduate School, 2Department of Gastroenterology, First Affiliated Hospital, Jinzhou Medical University, Jinzhou 121000, Chin)

机构地区：[1]锦州医科大学研究生院,辽宁锦州121000 [2]锦州医科大学附属第一医院消化内科,辽宁锦州121000

出　　处：《细胞与分子免疫学杂志》2018年第1期59-64,共6页Chinese Journal of Cellular and Molecular Immunology

基　　金：辽宁省自然科学基金(201602280)

摘　　要：目的研究土槿乙酸(PAB)阻滞HepG2肝癌细胞周期并抑制其侵袭转移的机制。方法采用四甲基偶氮唑蓝(MTT)法观察PAB对人肝癌HepG2细胞增殖的影响;流式细胞术检测其对HepG2肝癌细胞周期的影响;免疫荧光细胞化学染色法检测PAB对HepG2肝癌细胞内α微管蛋白(α-tubulin)聚合形态及表达的影响;TranswellTM小室侵袭实验和划痕愈合实验检测PAB对肝癌HepG2细胞侵袭和迁移能力的影响;Western blot法检测细胞内α-tubulin、上皮钙黏蛋白(E-cadherin)、基质金属蛋白酶9(MMP-9)的蛋白水平。结果 PAB可剂量依赖性地抑制HepG2细胞增殖并将细胞阻滞于G2/M期;PAB可显著改变微管蛋白聚合态使其表达明显减弱,PAB处理的HepG2细胞侵袭和迁移能力明显降低;细胞内α-tubulin、MMP-9蛋白水平降低,E-cadherin蛋白水平增加。结论 PAB可以通过下调α-tubulin水平影响其聚合,将HepG2细胞周期阻滞于G2/M期,抑制HepG2肝癌细胞增殖,通过下调细胞骨架α-tubulin、MMP-9,上调E-cadherin,抑制HepG2肝癌细胞的侵袭和迁移。Objective To investigate the mechanisms of pseudolaric acid B （PAB） blocks cell cycle and inhibits invasion and migration in human hepatoma HepG2 cells. Methods The proliferation effect of PAB on HepG2 cells was evaluated by MTT assay. The effect of PAB on the cell cycle of HepG2 cells was analyzed by flow cytometry. Immunofluorescence cytochemical staining was applied to observe the effect of PAB on the α-tubulin polymerization and expression in HepG2 cells. TranswellTM chamber invasion assay and wound healing assay were performed to detect the influence of PAB on the migration and invasion ability of HepG2 cells. Western blotting was used to determine the expressions of α-tubulin, E-cadhedn and MMP-9 in HepG2 cells after treated with PAB. Results PAB inhibited the proliferation of HepG2 cells in a dose-dependent manner and blocked the cell cycle in G2/M phase. PAB significantly changed the polymerization and decreased the expression of α-tubulin. The capacities of invasion and migration of HepG2 cells treated by PAB were significantly depressed. The protein levels of α-tubulin and MMP-9 decreased while the E-cadherin protein level increased. Conclusion PAB can inhibits the proliferation of HepG2 cells by down-regulating the expression of α-tubulin and influencing its polymedzation, arresting HepG2 cells in G2/M phase. Meanwhile, PAB also can inhibit the invasion and migration of HepG2 cells by lowering cytoskeleton α-tubulin and MMP-9, and increasing E-cadherin.

关 键 词：土槿乙酸 肝癌HEPG2细胞 细胞增殖 细胞周期 侵袭 迁移 

分 类 号：Q279[生物学—细胞生物学] R285.5[医药卫生—中药学]