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白花蛇舌草对人胃癌细胞系MNK-45线粒体膜电位及凋亡相关基因表达的影响

[Effects of hedyotis diffusa on mitochondrial membrane potential and expressions of apoptosis-related genes in human gastric cancer cell line MNK-45].

作者信息

DU Yang, Shao Shu-Li, Jiao Kai-He, Liu Xiang-Lu, Feng Yuan, Zhang Wei-Wei

机构信息

College of Life Sciences, Agriculture and Forestry, Qiqihar University, Qiqihar 161006, China.

Heilongjiang Provincial Key Laboratory of Resistance Gene Engineering and Protection of Biodiversity in Cold Areas, Qiqihar University, Qiqihar 161006, China.

出版信息

Zhongguo Ying Yong Sheng Li Xue Za Zhi. 2020 Mar;36(2):171-175. doi: 10.12047/j.cjap.5956.2020.038.

Abstract

To investigate the effects of hedyotis diffusa (injection) on mitochondrial membrane potential and expressions of apoptosis-related genes in human gastric cancer cell line MNK-45 cells. The human gastric cancer MNK-45 cells were divided into 4 groups, each group was set with 3 replicates. The control group was MNK-45 cells without added hedyotis diffusa; the 3 groups of experimental groups were treated with hedyotis diffusa at final concentrations of 20 , 30, 40 μg / ml respectively; each group was incubated for 48 h in a 5% carbon dioxide incubator, and the morphological changes of the cells were observed under a laser confocal microscope. Mitochondrial membrane potential was detected by flow cytometry. The expressions of (), and genes and proteins were detected by qRT-PCR and Western blot respectively. Compared with the control group, the mitochondrial membrane potentials of MNK-45 cells were significantly reduced in the hedyotis diffusa treated groups at final concentrations of 20, 30, and 40 μg / ml (<0. 01). The gene expressions of , , and were significantly up-regulated (<0. 01) and their protein expressions were also significantly increased (<0. 05 or <0. 01). The 40 μg / ml hedyotis diffusa treatment group performed best. In the final concentration range of 20 ~ 40 μg / ml, hedyotis diffusa can reduce human gastric cancer MNK-45 cells mitochondrial membrane potential, induce apoptosis and up-regulate Cyt c, caspase3 and caspase9 gene expressions.

摘要

探讨白花蛇舌草(注射液)对人胃癌细胞系MNK-45细胞线粒体膜电位及凋亡相关基因表达的影响。将人胃癌MNK-45细胞分为4组,每组设3个复孔。对照组为未添加白花蛇舌草的MNK-45细胞;3组实验组分别用终浓度为20、30、40μg/ml的白花蛇舌草处理;每组在5%二氧化碳培养箱中孵育48 h,在激光共聚焦显微镜下观察细胞形态变化。采用流式细胞术检测线粒体膜电位。分别采用qRT-PCR和Western blot检测()、和基因及蛋白的表达。与对照组相比,终浓度为20、30、40μg/ml的白花蛇舌草处理组MNK-45细胞的线粒体膜电位显著降低(<0.01)。、和的基因表达显著上调(<0.01),其蛋白表达也显著增加(<0.05或<0.01)。40μg/ml白花蛇舌草处理组效果最佳。在20~40μg/ml的终浓度范围内,白花蛇舌草可降低人胃癌MNK-45细胞线粒体膜电位,诱导细胞凋亡,并上调Cyt c、caspase3和caspase9基因表达。

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