摘要
目的:研究去甲斑蝥素对人卵巢癌SKOV3细胞的细胞毒作用及作用机制。方法:MTT法检测去甲斑蝥素对细胞增殖的影响,倒置显微镜和荧光显微镜观察细胞的形态学变化,流式细胞术检测细胞凋亡和活性氧(reactive oxygen species,ROS)的水平,蛋白印迹法检测相关蛋白的表达。结果:去甲斑蝥素能够时间和剂量依赖性地抑制SKOV3细胞生长。52μmol/L的去甲斑蝥素作用12~48 h后可使细胞核皱缩、染色质凝聚,形成明显的凋亡小体;还可时间依赖性地诱导细胞凋亡、G2/M期周期阻滞以及活性氧的产生。活性氧清除剂NAC可以显著降低去甲斑蝥素诱导的活性氧的产生,抑制细胞凋亡和G2/M期周期阻滞(P<0.05)。5 mmol/L NAC抑制去甲斑蝥素引起的Bax和p21表达上调,增加去甲斑蝥素引起的procaspase-3和Bcl-2的表达降低。结论:去甲斑蝥素通过升高活性氧的水平介导SKOV3细胞凋亡和G2/M期周期阻滞。
Objective: The study was aimed to investigate the cytotoxicity and mechanisms of norcantharidin in SKOV3 cells. Methods: MTT assay was performed to assess the inhibition ofnorcantharidinin SKOV3 cells. The morphological changes were examined under inverted microscope and fluorescence microscope.The cell cycle alteration, apoptosis and ROS generation were detected by flowcytometry. The expression levels of protein were examined by Western blotting. Results: Norcantharidin could inhibit SKOV3 cell proliferation in a time-and concentration-dependent manner. Remarkable morphologic changes were observed with treatment of 52 μmol/L norcantharidin for 12~48 h, including cell nuclear shrinkage, DNA condensation and apoptotic bodies. Norcantharidin also induced G2/M cell cycle arrest, increasedapoptosis and ROS level in a time-dependent manner.Besides,the ROS level, apoptosis and G2/Mcell cycle arrest of norcantharidin-treated cases were markedly decreased by the ROS scavenger NAC(P<0.05). After pretreated by 5 mmol/L NAC, enhancement of Bax, p21 expression and attenuation of procaspase-3 and Bcl-2 expression were thoroughly reversed compared with norcantharidin-treated case. Conclusion: Norcantharidin exhibited growth inhibition in SKOV3 cells, induced apoptosis and G2/Mcell cycle arrest by increasing the ROS level.
作者
董秀
冯晓丹
王英
韩兆丰
王晓波
李海波
DONG Xiu;FENG Xiaodan;WANG Ying;HAN Zhaofeng;WANG Xiaobo;LI Haibo(Liaoning University of Traditional Chinese Medicine,Shenyang 110847,Liaoning,China;Key Laboratory of Ministry of Education for TCM Viscera-State Theory and Applications,Liaoning University of Traditional Chinese Medicine,Shenyang 110847,Liaoning,China)
出处
《中华中医药学刊》
CAS
北大核心
2019年第8期1995-1999,I0023,I0024,共7页
Chinese Archives of Traditional Chinese Medicine
基金
辽宁省教育厅一般项目(L201603)
辽宁省科技厅博士科研启动基金项目(201501094)
沈阳市科技计划项目(F16-205-1-20)
