三叶青提取物对宫颈癌HeLa细胞增殖的影响及作用机制研究

Effect of Radix Tetrastigmae extracts on proliferation of HeLa cells and its mechanism

目的分析三叶青提取物对宫颈癌HeLa细胞增殖的影响,并探讨其作用机制。方法取对数生长期的HeLa细胞分为5组,分别加入0(对照组)、1、2、4、8mg/ml三叶青提取物。培养24或48h后,采用MTT法检测三叶青提取物对HeLa细胞增殖的影响;培养48h后于倒置显微镜下观察细胞形态,采用流式细胞仪检测细胞凋亡情况,采用Western blot法检测HeLa细胞迁移相关蛋白基质金属蛋白酶MMP-2、MMP-9和细胞凋亡相关蛋白p53、Bax的表达情况。结果三叶青提取物抑制HeLa细胞增殖,且具有浓度依赖性,当药物浓度达到8mg/ml时,24、48h细胞抑制率分别为38.4%、47.2%。三叶青提取物使HeLa细胞胞质浓缩、伪足缩短、体积变小等形态学改变。三叶青提取物通过诱导p53及Bax蛋白表达上调(4mg/ml组与对照组相比,p53与Bax表达水平分别为2.21倍和1.79倍,均P<0.05)促进细胞凋亡;且MMP-2、MMP-9蛋白表达下调,三叶青提取物通过该途径抑制HeLa细胞迁移。结论三叶青提取物对宫颈癌HeLa细胞增殖起抑制作用。其或通过上调p53及Bax蛋白表达诱导HeLa细胞凋亡,通过下调MMP-2及MMP-9表达影响HeLa细胞侵袭和迁移。

Objective To investigate the effects of Radix Tetrastigmae(RT)extracts on proliferation of HeLa cells and the underlying mechanism.Methods HeLa cells in logarithmic growth phase were treated with RT extracts of concentrations 0(control group),1,2,4,and 8mg/ml for 24 and 48h.MTT method was used to detect the effect of RT extracts on proliferation of HeLa cells.After 48h of incubation,cell morphology was observed under an inverted microscope,and cell apoptosis was detected by flow cytometry.Expression of matrix metalloproteinase MMP-2,MMP-9,and apoptosis-related proteins p53 and Bax in HeLa cells were detected by Western blot.Results RT extracts inhibited HeLa cell proliferation in a concentration-dependent manner.When the concentration of RT extracts reached 8mg/mL,the inhibiting rates were 38.4%and 47.2%after incubation for 24h and 48h,respectively.RT extracts induced morphological changes including cytoplasmic concentration,pseudo pod shortening and volume reduction of HeLa cells.Western blot results showed that RT extracts up-regulated p53 and Bax expression(2.21 and 1.79 folds when compared 4mg/mL group vs.control group,all P<0.05),and down-regulated MMP-2 and MMP-9 expression.Conclusion Radix Tetrastigmae extract has inhibitory effect on proliferation of HeLa cells.It may induce HeLa cell apoptosis by up-regulating p53 and Bax protein expression,and affect HeLa cell invasion and migration by down-regulating MMP-2 and MMP-9 expression.