丹参酮ⅡA对人肝癌HepG2细胞增殖和迁移的抑制作用及促凋亡作用

doi:10.13481/j.1671-587x.20190311

吉林大学学报(医学版) ›› 2019, Vol. 45 ›› Issue (03): 531-538.doi: 10.13481/j.1671-587x.20190311

• 基础研究 • 上一篇

丹参酮ⅡA对人肝癌HepG2细胞增殖和迁移的抑制作用及促凋亡作用

陈曦¹, 柏林², 王映映¹, 曹爽², 穆宏平², 王翊霖², 石浩², 张倩², 高鑫², 张成义², 张若文¹

1. 北华大学医学院病原生物学教研室, 吉林吉林 132011;
2. 北华大学药学院药理教研室, 吉林吉林 132011

收稿日期:2018-12-27 发布日期:2019-06-05
通讯作者: 张若文,副研究员,硕士研究生导师(Tel:0432-64608072,E-mail:zrw1828@163.com) E-mail:zrw1828@163.com
作者简介:陈曦(1968-),女,上海市人,教授,医学博士,主要从事中药毒理学方面的研究。
基金资助:
吉林省科技厅科技发展计划项目资助课题（20180101142JC）；吉林省教育厅"十三五"科技项目资助课题（JJKH20190662KJ）；国家级大学生创新创业训练计划项目资助课题（201711923009）

Inhibitory effect of tanshinone ⅡA on proliferation and migration of human liver cancer HepG2 cells and its apoptosis-promoting effect

CHEN Xi¹, BAI Lin², WANG Yingying¹, CAO Shuang², MU Hongping², WANG Yilin², SHI Hao², ZHANG Qian², GAO Xin², ZHANG Chengyi², ZHANG Ruowen¹

1. Department of Pathogenic Biology, College of Medical Sciences, Beihua University, Jilin 132011, China;
2. Department of Pharmacology, College of Pharmacy, Beihua University, Jilin 132011, China

Received:2018-12-27 Published:2019-06-05

摘要/Abstract

摘要： 目的：观察丹参酮ⅡA （Tan ⅡA）对人肝癌HepG2细胞增殖的抑制作用及诱发细胞发生迁移和凋亡作用，探讨其可能的作用机制。方法：常规体外培养人肝癌HepG2细胞，实验分为空白组和不同浓度Tan ⅡA组。不同浓度Tan ⅡA组分别加入终浓度为0.5、1.0、2.0、5.0和10.0 mg·L^-1Tan ⅡA，继续培养24 h。倒置显微镜观察各组HepG2细胞形态表观，MTT法检测各组HepG2细胞增殖抑制率，细胞划痕实验检测细胞迁移情况，RT-PCR法检测各组HepG2细胞中核转录因子κB （NF-κB）和基质金属蛋白酶9（MMP-9） mRNA的表达水平，流式细胞术检测各组不同细胞周期HepG2细胞百分比，TUNEL法检测各组HepG2细胞凋亡率。结果：不同浓度Tan ⅡA组细胞形态表观均发生改变。与空白组比较，1.0和2.0mg·L^-1Tan ⅡA组HepG2细胞体积缩小，连接疏散，生长状态差，1.0、2.0、5.0和10.0 mg·L^-1Tan ⅡA组HepG2细胞增殖抑制率明显升高（P<0.05），且呈剂量依赖性。细胞划痕实验检测，与空白组比较，2.0和5.0 mg·L^-1Tan ⅡA组HepG2细胞迁移数减少。RT-PCR法检测，与0.5 mg·L^-1Tan ⅡA组比较，1.0、2.0和5.0 mg·L^-1Tan ⅡA组HepG2细胞中MMP-9与NF-κB mRNA表达水平降低（P<0.05或P<0.01）。流式细胞术检测，与空白组比较，1.0、2.0、5.0和10.0 mg·L^-1Tan ⅡA组S期HepG2细胞百分比降低（P<0.05），G₀/G₁和G₂期细胞百分比升高（P<0.05）。TUNEL法检测，与空白组比较，0.5、1.0、2.0、5.0和10.0 mg·L^-1 Tan ⅡA组HepG2细胞凋亡率升高（P<0.05或P<0.01）。结论：不同浓度Tan ⅡA均可明显抑制人肝癌HepG2细胞的增殖与迁移，并诱发细胞凋亡，且呈一定的剂量依赖关系，其机制可能与抑制HepG2细胞中NF-κB和MMP-9 mRNA表达有关。

关键词: 丹参酮ⅡA, 肝肿瘤, 细胞迁移, 细胞凋亡, 核转录因子, 基质金属蛋白酶9

Abstract: Objective:To observe the inhibitory effect of tanshinone ⅡA (Tan ⅡA) on the proliferation of liver cancer HepG2 cells and its inductive effect on the migration and apoptosis of HepG2 cells, and to explore the possible mechanism. Methods:The human liver cancer HepG2 cells were cultured in vitro. The HepG2 cells were divided into blank group and different concentrations of Tan ⅡA groups. The cells in different concentrations of Tan ⅡA groups were added with Tan ⅡA at the final concentrations of 0.5, 1.0, 2.0, 5.0 and 10.0 mg·L^-1 Tan ⅡA and cultured for 24 h.The morphology of HepG2 cells in various groups was observed under inverted microscope. The inhibitory rates of proliferation of HepG2 cells in various groups were detected by MTT assay. The migration of HepG2 cells in various groups were evaluated by cell scratch assay. The expression levels of nuclear factor-κB (NF-κB) and matrix metalloproteinase-9 (MMP-9) mRNA were detected by RT-PCR method. Flow cytometry was used to detect the percentages of HepG2 cells at different cell cycles in various groups,and the apoptotic rates of HepG2 cells in various groups were detected by TUNEL method. Results:The morphology of HepG2 cells in different concentrations of Tan ⅡA groups were changed. Compared with blank group, the cells in 1.0 and 2.0 Tan ⅡA groups showed shrinkage,scattered connection and poor growth,and the inhibitory rates of proliferation of HepG2 cells in 1.0, 2.0, 5.0, and 10.0 mg·L^-1 Tan ⅡA groups were significantly increased (P<0.05) in a dose-dependent manner.The cell scratch assay results showed that with the increasing of Tan ⅡA concentration, the migration number of the cells in 2.0 and 5.0 mg·L^-1 Tan ⅡA groups were decreased significantly. Compared with 0.5 mg·L^-1 TanⅡA group, the expression levels of MMP-9 and NF-κB mRNA in the HepG2 cells in 1.0,2.0,and 5.0 mg·L^-1 Tan ⅡA groups were decresed(P<0.05 or P<0.01). The flow cytometry results showed that compared with blank group, the percentages of HepG2 cells in S phase in 1.0,2.0,5.0, and 10.0 mg·L^-1 Tan ⅡA groups were decreased (P<0.05),and the percentages of HepG2 cells in G₀/G₁ and G₂ phases were increased(P<0.05); the TUNEL results showed that compared with blank group, the apoptotic rates of HepG2 cells in 0.5, 1.0, 2.0, 5.0, and 10.0 mg·L^-1 Tan ⅡA groups werer increased (P<0.05 or P<0.01). Conclusion:Different concentrations of Tan ⅡA could significantly inhibit the proliferation and migration of human liver cancer HepG2 cells, and induce the apoptosis in a dose-dependent manner; its mechnasim may be related to the inhibition of the expressions of NF-κB and MMP-9 mRNA.

Key words: tanshinone ⅡA, liver neoplasms, cell migration, apoptosis, nuclear factor-κB, matrix metalloproteinase-9

中图分类号:

R735.7

陈曦, 柏林, 王映映, 曹爽, 穆宏平, 王翊霖, 石浩, 张倩, 高鑫, 张成义, 张若文. 丹参酮ⅡA对人肝癌HepG2细胞增殖和迁移的抑制作用及促凋亡作用[J]. 吉林大学学报(医学版), 2019, 45(03): 531-538.

CHEN Xi, BAI Lin, WANG Yingying, CAO Shuang, MU Hongping, WANG Yilin, SHI Hao, ZHANG Qian, GAO Xin, ZHANG Chengyi, ZHANG Ruowen. Inhibitory effect of tanshinone ⅡA on proliferation and migration of human liver cancer HepG2 cells and its apoptosis-promoting effect[J]. Journal of Jilin University(Medicine Edition), 2019, 45(03): 531-538.

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丹参酮ⅡA对人肝癌HepG2细胞增殖和迁移的抑制作用及促凋亡作用

Inhibitory effect of tanshinone ⅡA on proliferation and migration of human liver cancer HepG2 cells and its apoptosis-promoting effect

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