渥曼青霉素联合丝裂霉素对人肝癌HepG-2细胞增殖和凋亡的影响.docVIP

渥曼青霉素联合丝裂霉素对人肝癌HepG-2细胞增殖和凋亡的影响 张健1 ，王洪林1，张潜1，赵媛2(400016重庆，重庆医科大学附属第一医院：肝胆外科1，眼科2) [摘要] 目的 研究渥曼青霉素联合丝裂霉素对肝癌HepG-2细胞增殖、凋亡及相关基因 Cyclin D1、Bad表达的影响。方法 MTT比色法检测细胞增殖状态；Hoechst 33258荧光染色观察凋亡细胞核形态学变化，流式细胞术检测肿瘤细胞周期和凋亡；逆转录聚合酶联反应及Western blot 检测PI3K/Akt信号传导通路相关基因和蛋白的表达。结果 联合用药组的细胞增殖抑制率显著高于单独用药组(P0.05)；联合用药作用24 h后流式细胞术显示细胞明显阻滞于G0/G1期；细胞凋亡率由处理前的（3.23±1.32）%提高到14.82±1.36）%(P0.01)；荧光显微镜下可见典型的细胞凋亡形态学改变；联合用药后细胞p-Akt活化程度降低，Cyclin D1表达量减少，Bad表达量增加（P0.05)。结论 渥曼青霉素与丝裂霉素联合应用，具有协同抗肿瘤效应。 [关键词] 肝癌；渥曼青霉素；丝裂霉素；凋亡；PI3K/Akt信号通路 [中图法分类号] R735.7 [文献标志码] A Effects of Wortmannin combined with mitomycin C on HepG-2 cell Zhang Jian, 1 Wang Hong-lin, 1 Zhang Qian 1 Zhao Yuan 2（Department of Hepatobiliary Surgery, First Affiliated Hospital , Chongqing Medical University, Chongqing 400016. 2 Department of Ophthalmology, First Affiliated Hospital,Chongqing Medical University, Chongqing 400016.） Abstract: Objective this study was to investigate the effects of Wortmannin combined use of mitomycin C on the proliferation,apoptosis and expressions of related CyclinD1.Bad genes in liver cancer.Methods Cell suppression rate were detected by MTT assay; The nucleus apoptosis of Hep-G-2 cells were examined by Hochest 33258; The change of cell cycle and apoptosis in HepG-2 cells was detected by flowcytometry (FCM),expression of PI3K/ Akt signaling transduction pathway related genes and proteins were tested by reverse transcription-polymerase chain reaction (RT-PCR) and Western Blot assay . Results The inhibition ratio of cell proliferation was significant higher in Combined use of drugs groups than in either single groups(P0.05); FCM show : HepG-2 cells were prohibited at G0/G1 phrase ,apoptosis rates of HepG-2 cells increased from （3.23±1.32）% to（14.82±1.36）% After incubation with Combined use of Wortmannin、MMC for 24 hours. (P 0.01).typical changes in apoptotic morphology of HepG-2 cells were observed by fluorescence mic-roscopy .activation of p-Akt was inh