三氧化二砷对骨肉瘤细胞株MG63／dox多药耐药的逆转作用及其机制  

Reversal effect and mechanism of arsenic trioxide on multidrug resistance of human osteosarcoma cells MG63 / dox

作　　者：丰涛[1] 沈赞[2] 

机构地区：[1]苏州大学附属儿童医院检验科,江苏苏州215025 [2]上海交通大学附属第六人民医院肿瘤内科,200233

出　　处：《临床肿瘤学杂志》2016年第1期6-11,共6页Chinese Clinical Oncology

基　　金：国家自然科学基金资助项目(81172105)

摘　　要：目的 探讨三氧化二砷（As2O3）对骨肉瘤阿霉素（ADM）耐药细胞株MG63/dox的逆转作用及其机制。方法 采用不同浓度ADM（1～1000 ng／ml）和As2O3（0.25～16 μmol／L）分别单独作用于MG63和MG63/dox细胞48 h，并选取2 μmol／L As2O3联合不同浓度ADM（1～1000 ng／ml）作用于MG63/dox细胞24、48、72 h，同时设不加任何药物处理的对照组。应用CCK-8法检测两种药物对MG63和MG63／dox细胞增殖的影响，流式细胞术检测2 μmol／L As2O3、200 ng／ml ADM单独或联合处理48 h后MG63／dox细胞的周期分布和凋亡率，蛋白免疫印迹法检测2 μmol／L As2O3、200 ng／ml ADM单独或联合处理48 h后MG63／dox细胞中P-gp、Bcl-2和Bax蛋白的表达水平。结果 As2O3和ADM单药作用于MG63/dox细胞48 h，细胞增殖均未受到影响；4、8、16 μmol/L As2O3和100、200、400、500、1000 ng/ml ADM分别作用于MG63细胞48 h，细胞增殖明显受到抑制（P＜0.05）。As2O3联合ADM抑制MG63／dox细胞增殖的作用明显高于ADM单药组和对照组（P＜0.05），且抑制作用呈时间依赖性。与对照组相比，两药联合处理组的细胞凋亡率升高，G0／G1期细胞比例降低，G2／M期细胞比例升高，以上差异均有统计学意义（P＜0.05）。两药联合处理组MG63／dox细胞中Bax蛋白表达水平明显升高，而P-gp和Bcl-2蛋白表达水平明显降低，与对照组比较，差异均有统计学意义（P＜0.05）。结论 As2O3联合ADM能够抑制骨肉瘤耐药细胞MG63／dox的增殖，这一作用可能与诱导细胞凋亡、上调Bax表达以及下调P-gp和Bcl-2表达有关。Objective To investigate the effect of arsenic trioxide（As2O3） on human osteosarcoma cells MG63 and drug resistant cells MG63/dox, and to explore its possible mechanism. Methods MG63 and MG63/dox cells were respectively treated with adriamycin （ ADM, 1-1000 ng/ml） or As2O3 （ 0. 25-16 μmol/L） for 48 h ; 2 μmol/L As2O3 in combination with ADM （ 1-1000 ng/ml） was employed to treat MG63/dox cells for 24, 48, 72 h. Meanwhile, negative control group was set. The effect of As2O3 and ADM on proliferation of MG63/dox cell were detected by CCK-8 assay. The cell cycle and apoptotic rate of MG63/dox cells after treatment with As2O3（2 Ixmol/L） and ADM（200 ng/ml） alone and their combination for 48 h were detected by flow cytometry, and the expression levels of P-gp, Bcl-2 and Bax proteins treated by As2O3（2 μmol/L） or ADM（200 ng/ml） alone or in combination for 48 h were ana- lyzed by Western blotting, respectively. Results As2O3 or ADM alone treated MG63/dox cells for 48 h, the proliferation of cells had no obvious changes. The concentration of 4, 8, 16 μmol/L As2O3 and 100, 200, 400, 500, 1000 ng/ml ADM respectively treated MG63 cells for 48 h, the cell proliferation was inhibited （P〈0. 05）. As2O3 in combination with ADM markedly inhibited cell prolifera- tion in MG63/dox cells compared with ADM group and control group（P〈0. 05）in time-dependent manner. Compared with the control group, As2O3 in combination with ADM increased the apoptosis of MG63/dox ceils and Gz/M phase cell cycle arrest, but reduced Go/ G1 phase cell arrest（P〈0. 05）. Furthermore, the expression of Bax protein was found to be up-regulated in MG63/dox cells, and the expressions of P-gp and Bcl-2 proteins were lower than those in control group（P〈0. 05）. Conclusion As2O3 combined with ADM can inhibit the proliferation of MG63/dox cells. This effect may associate with induction of apoptosis, up-regulation of Bax expression and down-regulation of P-gp and Bcl-2 expressions.

关 键 词：三氧化二砷(As2O3) 骨肉瘤 MG63细胞株 MG63/dox细胞株 多药耐药 凋亡 

分 类 号：R738.1[医药卫生—肿瘤]