盐霉素对人黑色素瘤M21细胞增殖及自噬流的影响

doi:10.11904/j.issn.1002-3070.2018.03.002

实用肿瘤学杂志 ›› 2018, Vol. 32 ›› Issue (3): 203-207.doi: 10.11904/j.issn.1002-3070.2018.03.002

盐霉素对人黑色素瘤M21细胞增殖及自噬流的影响

刘雅菁¹, 刘国钰¹, 王树滨², 方征宇¹

1.深圳北京大学香港科技大学医学中心(深圳 518036);
2.北京大学深圳医院肿瘤科

收稿日期:2018-04-10 出版日期:2018-07-10 发布日期:2018-07-10
通讯作者: 方征宇,E-mail:fangzy796@163.com
作者简介:刘雅菁,女,(1993-),硕士研究生,从事肿瘤耐药机制方面的研究
基金资助:
广东省自然科学基金(编号:2015A030313754);深圳市2017年课题(编号:JCYJ20170306161757367)

Effect of salinomycin on proliferation and autophagic flow of human malignant melanoma M21 cells

LIU Yajing¹,LIU Guoyu¹,WANG Shubin²,FANG Zhengyu¹

1.Institute of Biomedical Research,Shenzhen PKU-HKUST Medical Center,Shenzhen 518036,China;
2.Department of Cancer Surgery,Peking University Shenzhen Hospital

Received:2018-04-10 Online:2018-07-10 Published:2018-07-10

摘要/Abstract

摘要： 目的探讨盐霉素对人黑色素瘤M21细胞增殖及自噬流的影响。方法采用MTS法检测细胞生存率,计算半数抑制浓度IC₅₀值。观察给药后细胞形态学变化。采用流式细胞仪检测细胞的凋亡率。Western blot检测给药后M21细胞内自噬相关蛋白LC3B表达情况。透射电镜下观察给药后M21细胞内是否有自噬小体产生。Western blot和免疫荧光检测p62蛋白水平和定位变化。结果盐霉素对人黑色素瘤M21细胞具有明显的增殖抑制作用,IC₅₀值为(1.38±0.18)μM。给药后细胞增殖速度减慢,同时细胞中出现明显的空泡。盐霉素不能促进细胞发生凋亡,但能使细胞内LC3B-Ⅱ/LC3B-Ⅰ的比值逐渐增加。在透射电镜下观察到自噬小体的增多与堆积。盐霉素作用后p62蛋白水平些许升高,逐渐在胞质内聚集成团状,表明自噬流被抑制。结论盐霉素能够抑制人黑色素瘤M21细胞的增殖,其机制可能与诱导自噬小体增多、抑制自噬流有关。

关键词: 黑色素瘤, M21细胞株, 盐霉素, 自噬

Abstract: Objective The objective of this study was to investigate the effect of salinomycin on proliferation and autophagic flux of human melanoma M21 cells.Methods The cell survival rate was determined by MTS assay and IC₅₀ values(half inhibitory concentration)were calculated.The morphological changes of cells after salinomycin administration were observed under optical microscope.Flow cytometry was used to examine the apoptosis rate of M21 cells.Western blot was used to detect the expression of autophagic-related protein LC3B and p62 in M21 cells.The presence of autophagosomes in M21 cells after salinomycin administration was observed under transmission electron microscope.Western blot and immunofluorescence were used to detect the level of p62 protein and localizing changes in M21 cells.Results Salinomycin significantly inhibited proliferation of M21 cells,and the IC₅₀ values were(1.38±0.18)μM.After salinomycin administration,the proliferation rate of M21 cells was slowed down,and obvious vacuoles appeared in the cells.Salinomycin could not only induce cell apoptosis,but it also increased the ratio of LC3B-Ⅱ/LC3B-Ⅰ in M21 cells.The increase and accumulation of autophagosomes were directly observed under transmission electron microscope.The level of p62 protein was slightly elevated after salinomycin treatment and gradually aggregated into the cytoplasm,indicating that autophagic flux was inhibited.Conclusion Salinomycin can inhibit the proliferation of human malignant melanoma M21 cells,and its mechanism may be related to the accumulation autophagosomes granules and inhibition of autophagic flux.

Key words: Malignant melanoma, M21 cell line, Salinomycin, Autophagy

中图分类号:

R739.5

刘雅菁, 刘国钰, 王树滨, 方征宇. 盐霉素对人黑色素瘤M21细胞增殖及自噬流的影响[J]. 实用肿瘤学杂志, 2018, 32(3): 203-207.

LIU Yajing, LIU Guoyu, WANG Shubin, FANG Zhengyu. Effect of salinomycin on proliferation and autophagic flow of human malignant melanoma M21 cells[J]. PRACTICAL ONCOLOGY JOURNAL, 2018, 32(3): 203-207.

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盐霉素对人黑色素瘤M21细胞增殖及自噬流的影响

Effect of salinomycin on proliferation and autophagic flow of human malignant melanoma M21 cells

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