桦褐孔菌醇诱导人卵巢癌SKOV3细胞株凋亡及其分子机制研究  

Effect of Inotodiol on Cell Proliferation and Apoptosis of Human Ovary Cancer Cell Line SKOV3 and Its Molecular Mechanism

作　　者：钟秀宏[1] 赵丽微[1] 赵芬琴[2] 杨淑艳[1] 徐野[1] 任旷[1] 钟越[1] 叶龙[1] 白杨[1] 杨宁江[3] 

机构地区：[1]吉林医药学院,吉林吉林132013 [2]河南大学药学院,河南开封475004 [3]吉林医药学院附属医院,吉林吉林132011

出　　处：《中国药学杂志》2014年第3期191-194,共4页Chinese Pharmaceutical Journal

基　　金：吉林省自然科学基金项目(201215226);吉林省卫生厅项目(2011z062);吉林医药学院大学生科研基金资助项目(吉医学科字[2011]第03号)

摘　　要：目的研究桦褐孔菌醇(inotodiol)诱导人卵巢癌细胞SKOV3凋亡及其对相关蛋白Bcl-2、Bax和caspase-3表达的影响。方法选用人卵巢癌SKOV3细胞株进行体外培养,四甲基偶氮唑蓝法检测桦褐孔菌醇对人卵巢癌SKOV3细胞增殖的影响;应用透射电镜观察细胞形态学变化;通过TUNEL染色测定凋亡率;通过免疫细胞化学法检测凋亡相关蛋白Bcl-2,Bax和caspase-3的表达。结果桦褐孔菌醇抑制人卵巢癌细胞增殖,呈时间-剂量依赖性;形态学检测出现典型的细胞凋亡改变。50mg·L-1桦褐孔菌醇作用人卵巢癌SKOV3细胞24、48 h的凋亡率分别为(21.91±2.84)%和(43.83±3.22)%,肿瘤细胞凋亡率(AR)随药物作用时间的延长而上升,显示明显的时效关系(P<0.05)。免疫细胞化学法染色显示,桦褐孔菌醇组人卵巢癌SKOV3细胞Bcl-2基因蛋白的阳性表达率明显降低,而Bax、caspase-3基因蛋白阳性表达率明显升高。结论桦褐孔菌醇对人卵巢癌SKOV3细胞株有抗增殖和诱导凋亡作用,该药可能是通过下调凋亡抑制基因Bcl-2的表达和上调促进凋亡基因Bax的表达及启动caspase凋亡信号而发挥抗癌作用。OBJECTIVE To investigate the effect of inotodiol in inducing apoptosis of human ovary cancer cell line SKOV3, and its influence on the related apoptosis proteins such as Bcl-2, Bax and caspase-3. METHODS Human ovary cancer SKOV3 cells were cultured in vitro. Cytotoxcity of inotodiol was determined by MTT assay. Morphological changes of apoptotic cells were observed by transmission electron microscope（TEM）. Apoptotic rates of SKOV3 cells were confirmed by TUNEL（terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling）assay. The expressions of Bcl-2, Bax and caspase-3 were detected by immunocytochemical staining method. RESULTS Inotodiol can inhibit the SKOV3 cells growth in a time-and dose-dependent manner. After treatment with inotodiol, SKOV3 cells showed typical apoptotic features under TEM. After human ovary cancer cell line SKOV3 were treated with inotodiol in concentration of 50 mg稬-1for 24 and 48 h, their apoptotic rates were（21.91？2.84）% and（43.83？3.22）%, respectively. The apoptotic rate of SKOV3 cells induced by inotodiol increased in a time-dependent manner（P〈0.05）. Immunocytochemical staining showed that the expression of Bcl-2 decreased obviously, whereas those of Bax and caspase-3 increased significantly in inotodiol group, compared with the control group. CONCLUSION Inotodiol can inhibit the proliferation and induce the apoptosis of human ovary cancer cell line SKOV3 through up-regulation of Bax protein, down-regulation of Bcl-2 protein and activation of caspase apoptosis signal.

关 键 词：桦褐孔菌醇 卵巢癌 凋亡 

分 类 号：R965[医药卫生—药理学]