米托蒽醌通过活性氧诱导大鼠肝癌RH35细胞凋亡的研究

doi:10.16431/j.cnki.1671-7236.2016.12.024

《中国畜牧兽医》 ›› 2016, Vol. 43 ›› Issue (12): 3245-3250.doi: 10.16431/j.cnki.1671-7236.2016.12.024

米托蒽醌通过活性氧诱导大鼠肝癌RH35细胞凋亡的研究

张亚宏, 甘莹, 齐建国, 王建红

河南大学, 天然药物与免疫工程重点实验室, 开封 475004

收稿日期:2016-08-26 出版日期:2016-12-20 发布日期:2016-12-22
通讯作者: 齐建国, 王建红 E-mail:qijianguo@henu.edu.cn;jhworg@126.com
作者简介:张亚宏(1982-),女,山东济宁人,博士,副教授,研究方向:毒理学与肿瘤药理学,E-mail:zhangyahong_131@163.com
基金资助:
国家自然科学基金（U1404819）；河南省自然科学基金（132300410228）

Mitoxantrone Induced Apoptosis through ROS in Rat Hepatama RH35 Cell Line

ZHANG Ya-hong, GAN Ying, QI Jian-guo, WANG Jian-hong

Key Laboratory of Natural Medicine and Immuno-engineering, Henan University, Kaifeng 475004, China

Received:2016-08-26 Online:2016-12-20 Published:2016-12-22

摘要/Abstract

摘要：

试验旨在探讨米托蒽醌（mitoxantrone，MTN）对大鼠肝癌RH35细胞毒性及其作用机制。以MTT法检测MTN的细胞毒性，显微镜观察细胞形态变化，流式细胞仪检测细胞凋亡率及细胞中活性氧（reactive oxygen species，ROS）的产生，Western blotting检测相关蛋白的表达。结果显示，MTN时间和剂量依赖性地抑制RH35细胞的生长；15 μmol/L MTN作用于细胞24、48 h后可使细胞皱缩、变圆，并出芽形成明显的凋亡小体，且其可时间依赖性地诱导凋亡细胞比率的增加及ROS的产生；ROS清除剂NAC可以极显著降低MTN诱导的RH35细胞的ROS的产生和凋亡率（P<0.01）；15 mmol/L NAC可以下调MTN诱导的caspase-3、Bax及CytC表达增加，上调MTN诱导的Bcl-2表达降低。结果提示，MTN通过增加细胞内ROS而诱导RH35细胞发生凋亡。

关键词: 米托蒽醌; 凋亡; RH35细胞; 活性氧

Abstract:

The assay was aimed to study the cytotoxicity and mechanisms of mitoxantrone (MTN) in rat hepatama cell line RH35. MTT assay was performed to assess the cytotoxicity of MTN, and microscope was used to observe cellular morphologic changes. Apoptotic ratio and intracellular ROS generation were measured by flow cytometric analysis. The protein expression was examined by Western blotting analysis. The results showed that MTN inhibited RH35 cell growth in a time-and concentration-dependent manner. The morphologic changes were observed in the cells, including cell shrinkage, membrane blebbing and apoptotic bodies when treated with 15 μmol/L MTN for 24 and 48 h. And MTN also induced the increase of apoptotic ratio and generation of ROS in a time dependent manner. In addition, the intracellular ROS generation ratio and the apoptotic ratio of MTN treated group were extremely decreased by the ROS scavenger NAC (P<0.01). The pretreatment of RH35 cells with 15 mmol/L NAC thoroughly reversed the MTN-induced enhancement of caspase-3, Bax and CytC level and attenuation of Bcl-2 level. In conclusion, MTN induced apoptosis in RH35 cells through increasing the generation of ROS.

Key words: mitoxantrone; apoptosis; RH35 cell; ROS

中图分类号:

Q813.5

张亚宏, 甘莹, 齐建国, 王建红. 米托蒽醌通过活性氧诱导大鼠肝癌RH35细胞凋亡的研究[J]. 《中国畜牧兽医》, 2016, 43(12): 3245-3250.

ZHANG Ya-hong, GAN Ying, QI Jian-guo, WANG Jian-hong. Mitoxantrone Induced Apoptosis through ROS in Rat Hepatama RH35 Cell Line[J]. , 2016, 43(12): 3245-3250.

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米托蒽醌通过活性氧诱导大鼠肝癌RH35细胞凋亡的研究

Mitoxantrone Induced Apoptosis through ROS in Rat Hepatama RH35 Cell Line

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