【摘要】 目的:观察南赤瓟醇提物对宫颈癌HeLa细胞增殖与凋亡作用的影响,并探讨其可能的作用机制。方法:以0.05～1.00 mg/ml南赤瓟醇提物分别处理人宫颈癌HeLa细胞24、48、72 h后,以四甲基偶氮唑蓝比色法测定南赤瓟醇提物对HeLa细胞的生长抑制作用,并在倒置相差显微镜下进行一般形态学观察;用0.20和0.50mg/ml的南赤瓟提物处理HeLa细胞48h后,Annexin V-FITC/PI染色测定细胞凋亡率,Hoechst33258染色观察细胞形态变化,荧光定量PCR法检测凋亡相关基因Bcl-2、Bax、Caspase-9、Caspase-3mRNA的表达。结果:不同浓度(0.05～1.00 mg/ml)的南赤瓟醇提物具有抑制宫颈癌HeLa细胞增殖、促进细胞凋亡的作用,呈明显的时间-剂量依赖性;南赤瓟醇提物(0.20和0.5mg/ml)处理能显著抑制HeLa细胞增殖,促进其凋亡(P<0.01);上调Bax,Caspase-9和Caspase-3 mRNA表达,下调Bcl-2 mRNA表达和Bcl-2与Bax的比值(P<0.05或P<0.01)。结论:南赤瓟醇提物可抑制宫颈癌HeLa细胞增殖,促进细胞凋亡,其机制可能与上调Bax、Caspase-9、Caspase-3和下调Bcl-2凋亡相关基因表达有关。更多还原

【Abstract】 Objective:To investigate the effect of alcohol extraction from Thldiantha nudiflora Hemsl on proliferation and apoptosis of cervix cancer HeLa cells,and to explore the possible mechanism.Methods: HeLa cells were treated with different concentration of alcohol extraction from Thldiantha nudiflora Hemsl(0.05 ～ 1.00mg/ml) at different times(24,48,72 h),Cell viability was determined by MTT assay,and morphological changes of HeLa cells were visualized under reversal phase microscope;After HeLa cells were treated with alcohol extraction from Thldiantha nudiflora Hemsl(0.20 and 0.50 mg/ml),Cell morphology was examined by hoechst33258 through fluorescence microscope;Annexin V/PI double-staining was performed to detect apoptosis of Hela cells by flow cytometry;The Expressions of Bcl-2,Bax,Caspase-9 and Caspase-3 mRNA were detected by Real-time PCR,and Bcl-2/Bax ratio was analyzed.Results: Alcohol extraction from Thldiantha nudiflora Hemsl inhibited the proliferation and induced apoptosis of HeLa cells on dose-and time-depending manner,and the differences among different concentration and time groups were significant(P < 0.05,P < 0.01,respectively);Alcohol extraction from Thldiantha nudiflora Hemsl(0.20 and 0.50 mg/ml) might significantly inhibit proliferation,promote apoptosis(P < 0.05,P < 0.01,respectively);up-regulate the mRNA expression levels of Bax,Caspase-9 and Caspase-3,down-regulate expression level of Bcl-2 and Bcl-2/Bax ratio compared with the control group(P < 0.05,P < 0.01,respectively).Conclusion: Alcohol extraction from Thldiantha nudiflora Hemsl could effectively inhibit proliferation and induce apoptosis in cervical cancer HeLa cells.The mechanism may be related to regulating the expressions of the apoptosisrelated genes Bcl-2,Bax and Caspase-9,Caspase-3.更多还原