双硫仑联合顺铂对三阴性乳腺癌MDA-MB-231细胞的增殖抑制作用及其机制

doi:10.13481/j.1671-587x.20200316

摘要/Abstract

摘要： 目的：观察双硫仑（DSF）和顺铂（CDDP）联合使用对三阴性乳腺癌（TNBC） MDA-MB-231细胞的增殖抑制作用，并探讨其可能机制。方法：将MDA-MB-231细胞分为空白对照组、2μmol·L^-1DSF组、60μmol·L^-1CDDP组和联合组（60μmol·L^-1CDDP+1、2和4μmol·L^-1DSF）。MTT法检测各组MDA-MB-231细胞的存活率，流式细胞术检测各组MDA-MB-231细胞凋亡率，流式细胞术检测各组MDA-MB-231细胞中活性氧（ROS）水平；电感耦合等离子体质谱仪（ICP-MS）检测各组MDA-MB-231细胞中铂（Pt）水平。结果：MTT法和流式细胞术检测，作用72 h后，与60 μmol·L^-1 CDDP组比较，联合1组（60 μmol·L^-1CDDP+1 μmol·L^-1DSF）、联合2组（60 μmol·L^-1CDDP+2 μmol·L^-1DSF）和联合3组（60 μmol·L^-1CDDP+4 μmol·L^-1 DSF） MDA-MB-231细胞存活率降低（P<0.05）。作用24 h后，与60 μmol·L^-1 CDDP组和2 μmol·L^-1 DSF组比较，联合组（60 μmol·L^-1CDDP+2 μmol·L^-1 DSF） MDA-MB-231细胞凋亡率升高（P<0.05）。流式细胞术检测，与CDDP组比较，联合组（60 μmol·L^-1CDDP+2 μmol·L^-1 DSF） MDA-MB-231细胞中ROS水平升高（P<0.05）。ICP-MS检测，与CDDP组比较，联合组（20 μmol·L^-1CDDP+0.625 μmol·L^-1 DSF） TNBC MDA-MB-231细胞中Pt水平升高（P<0.05）。结论：DSF与CDDP联合使用能明显抑制MDA-MB-231细胞的增殖，其可能机制是通过提高MDA-MB-231细胞中ROS水平和Pt水平抑制肿瘤细胞生长。

关键词: 三阴性乳腺癌, MDA-MB-231细胞, 顺铂, 双硫仑, 活性氧

Abstract: Objective: To observe the inhibitory effect of combination of disulfiram (DSF) and cisplatin (CDDP) on the proliferation of triple-negative breast cancer (TNBC) MDA-MB-231 cells, and to explore its possible mechanism. Methods: The MDA-MB-231 cells in the logarithmic phase were divided into blank control group, DSF group, CDDP group and combination groups (60μmol·L^-1 CDDP+1,2 and 4μmol·L^-1DSF). MTT method was used to detect the survival rates of the MDA-MB-231 cells in various groups and flow cytometry was used to detect the apoptotic rates of the MDA-MB-231 cells in various groups, flow cytometry was used to detect the level of active oxygen(ROS)in the MDA-MB-231 cells in various groups, and Inductively Coupled Plasma-MassSpectrometry (ICP-MS) was used to detect the levels of Pt in the MDA-MB-231 cells in various groups. Results: The MTT method and flow cytometry results showed that after treated for 72 h, compared with 60μmol·L^-1 CDDP group,the survival rates of the MDA-MB-231 cells in combination 1 group (60μmol·L^-1CDDP+1μmol·L^-1DSF), combination 2 group (60μmol·L^-1CDDP+2μmol·L^-1DSF) and combination 3 group (60μmol·L^-1CDDP+4μmol·L^-1DSF) were decreased(P<0.05). After treated for 24 h, compared with 60μ mol·L^-1CDDP group and 2μ mol·L^-1DSF group, the apoptotic rate of the MDA-MB-231 cells in combination group (60μmol·L^-1CDDP+2μmol·L^-1 DSF) was increased (P<0.05). Compared with CDDP group, the level of ROS in the MDA-MB-231 cells in combination group(60μmol·L^-1CDDP+2μmol·L^-1 DSF) was increased (P<0.05).The ICP-MS results showed that compared with CDDP group, the level of Pt in the MDA-MB-231 cells in combination group(20μmol·L^-1CDDP+0.625μmol·L^-1 DSF) was increased (P<0.05). Conclusion: The combination of DSF and CDDP can significantly inhibit the proliferation of MDA-MB-231 cells,and its possible mechanism may be related to inhibiting the growth of tumor cells by increasing the ROS level and Pt content in the MDA-MB-231 cells.

Key words: triple-negative breast cancer, MDA-MB-231 cells, cisplatin, disulfiram, reactive oxygen species

中图分类号:

R737.9

文庆一, 焦苗苗, 谢潇, 彭敬予, 董凤歌, 魏雪, 杨明. 双硫仑联合顺铂对三阴性乳腺癌MDA-MB-231细胞的增殖抑制作用及其机制[J]. 吉林大学学报(医学版), 2020, 46(03): 523-529.

WEN Qingyi, JIAO Miaomiao, XIE Xiao, PEN Jingyu, DONG Fengge, WEI Xue, YANG Ming. Inhibitory effect of disulfiram combined with cisplatin on triple negative breast cancer MDA-MB-231 cells and its mechanism[J]. Journal of Jilin University(Medicine Edition), 2020, 46(03): 523-529.

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