左旋棉酚联合低浓度阿霉素诱导纤维肉瘤细胞凋亡及其机制  

Apoptosis induced by levo-gossypol combined with low concentration adriamycin in HT1080 cells and its mechanism

作　　者：张昭军[1] 周洋[1] 阿尔孜古丽.吐尔逊 锡林宝勒日 刘朝政[1] 张顺利[1] 

机构地区：[1]新疆医科大学附属肿瘤医院骨与软组织科,乌鲁木齐830011 [2]新疆医科大学第一附属医院医学研究中心,乌鲁木齐830011

出　　处：《重庆医学》2013年第20期2372-2375,共4页Chongqing medicine

摘　　要：目的研究左旋棉酚联合低浓度阿霉素(ADM)诱导纤维肉瘤HT1080细胞凋亡及其作用机制。方法采用噻唑蓝(MTT)检测药物对细胞的抗增殖作用,应用瑞氏-姬姆萨(WG)染色、赫斯特33258(hoechst 33258)染色、透射电镜观察细胞的形态学变化,流式细胞仪检测细胞的凋亡率和细胞周期。蛋白免疫印迹法(Western blotting)检测Bcl-2、Bax蛋白的表达,比色分析法检测细胞半胱氨酸天冬氨酸蛋白酶(caspase)-3、caspase-9的活性变化。结果左旋棉酚与低浓度ADM对HT1080细胞具有抑制增殖作用,联合应用作用增强,有浓度与时间依赖性。药物作用后出现细胞凋亡的特征性改变,琼脂糖凝胶电泳观察到梯状带型,流式细胞仪检测发现两药能诱导肿瘤细胞凋亡,联合应用更强。左旋棉酚使细胞阻滞于G1期,ADM使细胞阻滞于S期,联合应用S期的细胞明显增加。细胞内Bcl-2蛋白表达减少,Bax蛋白表达增多,同时出现caspase-9、caspase-3的相继活化,联合应用变化更加明显。结论左旋棉酚与低浓度ADM对纤维肉瘤HT1080细胞有明显抑制增殖及诱导凋亡的作用,联合应用作用加强。Objective To investigate apoptosis inducted by levo-gossypol combined with low concentration adriamycin in HT1080 cells and its mechanisms. Methods The anti-proliferation of drugs on cells was detected by MTT assay. The changes of cells morphology were observed by Wright-Giemsa staining, Hoechst 33258 staining and transmission electron microscopy. The ap- optosis rate and cell cycle were detected by flow cytometry. And the protein expression of Bcl-2 and Bax in HT1080 cell were detec- ted by Western-Blot. The changes of caspase-3 and caspase-9 were detected by colorimetric assay. Results Levo-gossypol and low concentration adriamycin showed inhibition of HT1080 cells in a dose and time dependent. The combination had more obvious effect. Levo-gossypol and low concentration adriamycin induced apoptosis of HT1080 cell, as evidenced by typical morphological changes. Flow cytometry detected that the drugs induced apoptosis of HT1080 cell. The combination had more obvious apoptosis-in- ducing effect. Levo-gossypol could induce G~ arrest and adriamycin could induce S arrest. The combination had more obvious in- crease of S arrest. Levo-gossypol and low concentration adriamycin can result in decrease of Bcl-2 protein,increase of Bax protein, and activation of caspase-9 and caspase-3. The combination had more obvious changes. Conclusion Levo-gossypol and low concen- tration adriamycin have the effect of anti-proliferation and induction of apoptosis in HT1080 cell. The combination has more obvious effect.

关 键 词：左旋棉酚 阿霉素 HT1080 细胞凋亡 

分 类 号：R738[医药卫生—肿瘤]