金荞麦提取物对人宫颈癌Hela细胞增殖和诱导凋亡的影响  

Anti-proliferative and apoptosis-inducing activity of Rhizoma Fagopyri Cymosi Extract in cervical cancer Hela cells

作　　者：潘朝旺[1] 王伟[2] 祁学章[2] 杨家林[1] 周先云[1] Pan Chaowang;Wang Wei;Qi Xuezhang;Yang Jialin;Zhou Xianyun(Ezhou Polytechnic,Ezhou 436009;Ezhou Central Hospital,Ezhou 436009)

机构地区：[1]鄂州职业大学医学院,鄂州436000 [2]鄂州市中心医院,鄂州436000

出　　处：《中药药理与临床》2018年第5期96-100,共5页Pharmacology and Clinics of Chinese Materia Medica

基　　金：鄂州职业大学科研项目(2017YB06)

摘　　要：目的:探讨金荞麦提取物对人宫颈癌Hela细胞增殖、凋亡作用及可能机制。方法:经不同浓度金荞麦提取物(0.4、2、10、50、250 g/ml)处理体外培养的人宫颈癌Hela细胞,采用MTT法检测细胞增殖活性;经6 g/ml金荞麦提取物处理的Hela细胞,相差显微镜和荧光显微镜观察细胞形态,Annexin V-FITC/PI染色法检测细胞凋亡,琼脂糖凝胶电泳法检测DNA碎片,ELISA法检测Caspase-1、3、8、9活性,Western blot法检测PARP、细胞色素C含量、Bcl-2、Bax蛋白表达。结果:与空白对照组比较,金荞麦提取物浓度为0.2 g/ml开始起效,24h内明显抑制Hela细胞增殖,并呈时间和剂量依赖性;与空白对照组比较,6 g/ml金荞麦提取物处理的Hela细胞出现明显的核固缩、核碎裂,细胞凋亡率、DNA碎片明显增加;与金荞麦提取物组比较,金荞麦提取物与caspase抑制剂混合组DNA碎片明显减少,细胞成活率明显升高;与作用0h组比较,6 g/ml金荞麦提取物处理的Hela细胞PARP裂片、细胞色素C、Bax和Caspase-3、8、9明显增加,Bcl-2明显减少。结论:金荞麦提取物可抑制Hela细胞增殖、诱导细胞凋亡,其机制可能与通过促进诱导凋亡蛋白Bax表达和抑制抗凋亡蛋白Bcl-2表达,从而增加细胞色素C和Caspases有关。Objective:To investigate the effect and possible mechanism of Rhizoma Fagopyri Cymosi Extract(RFCE) on proliferation and apoptosis of human cervical carcinoma Hela cells.Methods:After cervical carcinoma Hela cells were incubated with different concentrations of RFCE(0.4,2,10,50,250 g/ml),the cell proliferation activity was examined by MTT assay.After Hela cells were incubated with 6 g/ml RFCE,the cell morphological changes were observed by phase-contrast microscope and fluorescence microscope,apoptotic cells were detected by Annexin V-FITC/PI apoptosis detection kit,DNA fragmentation was analysed by agarose gel electrophoresis,activity of Caspases-1,3,8,9 was detected by ELISA kits,Western blot was used to detect the PARP cleavage,cyt c release and level of Bcl-2 family protein.Results:Compared with control group,proliferation of Hela cells was obviously inhibited by RFCE from the dose of 0.2 g/ml,in dose-time dependence within 24 hours.After incubated with 6 g/ml RFCE,the cell nuclear condensation and fragmentation was exhibited,while the apoptosis rate and DNA fragmentation was significantly increased.Compared with RFCE group,the DNA fragmentation was significantly decreased and cell survival rate was significantly increased in the RFCE and caspase inhibitors mixed group.After incubated with 6 g/ml RFCE,the PARP cleavage,cyt c release,Bax level and Caspase-3,8,9 was significantly increased and the level of Bcl-2 significantly decreased,comparing with the 0 h group.Conclusion:RFCE was suggested to inhibit proliferation of Hela cell and induce apoptosis.The possible mechanisms is promoting the expression of apoptotic protein Bax and inhibiting the expression of anti-apoptotic protein Bcl-2 which may increase cyt c and Caspase.

关 键 词：金荞麦提取物 宫颈癌 凋亡 

分 类 号：R285.5[医药卫生—中药学]