Ji Chen-Feng, Ji Yu-Bin
Engineering Research Center of Natural Anticancer Drugs, Ministry of Education, Harbin University of Commerce, Harbin, Heilongjiang 150076, P.R. China ; Center of Research on Life Science and Environmental Science, Harbin University of Commerce, Harbin, Heilongjiang 150076, P.R. China.
Exp Ther Med. 2014 May;7(5):1317-1322. doi: 10.3892/etm.2014.1606. Epub 2014 Mar 6.
Numerous studies have demonstrated that podophyllotoxin and its derivatives exhibit antitumor effects. The aim of the present study was to investigate SGC-7901 cell apoptosis and the underlying mechanism induced by podophyllotoxin. SGC-7901 cells were treated with varying concentrations of podophyllotoxin. MTT assays and flow cytometry were used to evaluate the effects of podophyllotoxin on the proliferation and apoptosis of SGC-7901 cells, while fluorescence inverted microscopy was used to observe the morphology of SGC-7901 cells that had been dyed with Hoechst 33258. In addition, laser scanning confocal microscopy was used to analyze the mitochondrial membrane potential (MMP) of SGC-7901 cells dyed with Rhodamine 123. Western blotting was performed to analyze the expression levels of cytochrome (cyt-), caspase-9 and caspase-3 in the SGC-7901 cells. The results indicated that podophyllotoxin was capable of inhibiting growth and inducing the apoptosis of SGC-7901 cells in a dose-dependent manner, causing cell cycle arrest at the G/M phase. After 48 h of treatment, the apoptotic morphology of SGC-7901 cells was clear, exhibiting cell protuberance, concentrated cytoplasms and apoptotic bodies. Following 24 h of treatment, the MMP of the SGC-7901 cells decreased. In addition, after 48 h, the expression of cyt- was shown to be upregulated, while the expression levels of pro-caspase-9 and pro-caspase-3 in the SGC-7901 cells were shown to be downregulated. In conclusion, apoptosis can be induced in SGC-7901 cells by podophyllotoxin, potentially via a mitochondrial pathway, indicating that podophyllotoxin may be a potent agent for cancer treatment.
众多研究表明,鬼臼毒素及其衍生物具有抗肿瘤作用。本研究旨在探讨鬼臼毒素诱导SGC - 7901细胞凋亡及其潜在机制。用不同浓度的鬼臼毒素处理SGC - 7901细胞。采用MTT法和流式细胞术评估鬼臼毒素对SGC - 7901细胞增殖和凋亡的影响,同时用荧光倒置显微镜观察经Hoechst 33258染色的SGC - 7901细胞的形态。此外,用激光扫描共聚焦显微镜分析经罗丹明123染色的SGC - 7901细胞的线粒体膜电位(MMP)。进行蛋白质免疫印迹法分析SGC - 7901细胞中细胞色素c(cyt - )、半胱天冬酶 - 9和半胱天冬酶 - 3的表达水平。结果表明,鬼臼毒素能够抑制SGC - 7901细胞的生长并诱导其凋亡,且呈剂量依赖性,导致细胞周期停滞于G/M期。处理48小时后,SGC - 7901细胞的凋亡形态明显,表现为细胞突起、细胞质浓缩和凋亡小体。处理24小时后,SGC - 7901细胞的MMP降低。此外,48小时后,cyt - 的表达上调,而SGC - 7901细胞中前半胱天冬酶 - 9和前半胱天冬酶 - 3的表达水平下调。综上所述,鬼臼毒素可能通过线粒体途径诱导SGC - 7901细胞凋亡,表明鬼臼毒素可能是一种有效的癌症治疗药物。