【摘要】 目的:观察丹参酮ⅡA （TanⅡA）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TanⅡA组。不同浓度TanⅡA组分别加入终浓度为0.5、1.0、2.0、5.0和10.0mg·L^-1 TanⅡA,继续培养24h。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TanⅡA组细胞形态表观均发生改变。与空白组比较,1.0和2.0mg·L^-1 TanⅡA组HepG2细胞体积缩小,连接疏散,生长状态差,1.0、2.0、5.0和10.0mg·L^-1 TanⅡA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5.0mg·L^-1 TanⅡA组HepG2细胞迁移数减少。RT-PCR法检测,与0.5 mg·L^-1 TanⅡA组比较,1.0、2.0和5.0 mg·L^-1TanⅡA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10.0mg·L^-1 TanⅡA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^-1TanⅡA组HepG2细胞凋亡率升高（P<0.05或P<0.01）。结论:不同浓度TanⅡA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-9mRNA表达有关。更多还原

【Abstract】 Objective:To observe the inhibitory effect of tanshinoneⅡA（TanⅡA）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 TanⅡA groups were added with TanⅡA at the final concentrations of 0.5,1.0,2.0,5.0 and 10.0 mg·L^-1 TanⅡA 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 TanⅡA groups were changed.Compared with blank group,the cells in1.0 and 2.0 TanⅡA 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 TanⅡA 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 Ⅱ A 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 TanⅡA 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 TanⅡA 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 TanⅡA 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.更多还原