MST1R抑制剂BMS-777607对乳腺癌MCF-7细胞增殖的抑制和凋亡诱导作用

doi:10.13481/j.1671-587x.20200304

摘要/Abstract

摘要： 目的：探讨巨噬细胞刺激1受体（MST1R）抑制剂BMS-777607对乳腺癌MCF-7细胞增殖和凋亡的影响，并阐明其作用机制。方法：采用不同浓度BMS-777607处理乳腺癌MCF-7细胞，将细胞分为对照组（0 μmol·L^-1 BMS-777607组）和0.5、1.0、2.0、5.0、10.0、15.0及20.0 μmol·L^-1 BMS-777607组。采用MTT法检测各组MCF-7细胞增殖率，克隆形成实验检测各组MCF-7细胞存活率，EDU成像和EDU流式细胞术检测各组MCF-7细胞增殖率，Hoechst33342染色法检测各组MCF-7细胞凋亡形态表现，流式细胞术检测各组MCF-7细胞凋亡率，Western blotting法检测各组MCF-7细胞中ERK、p-ERK、Akt、p-Akt、PARP、Cleaved PARP、Bax、Caspase-3、Cleaved Caspase-3、Caspase-9和Cleaved Caspase-9蛋白表达水平。结果：MTT检测，与对照组比较，5和10 μmol·L^-1BMS-777607组MCF-7细胞增殖率升高（P<0.05或P<0.01）。克隆形成实验，与对照组比较，5和20 μmol·L^-1BMS-777607组MCF-7细胞存活率升高（P<0.05或P<0.01）。EDU掺入法检测，与对照组比较，10和20 μmol·L^-1BMS-777607组MCF-7细胞增殖率升高（P<0.05或P<0.01）。Hoechst33342荧光染色，对照组MCF-7细胞核淡染，10μmol·L^-1BMS-777607组MCF-7细胞核少部分浓染、明亮，20 μmol·L^-1BMS-777607组MCF-7细胞核大部分浓染，细胞核染色质固缩、明亮。双染流式细胞术检测，与对照组比较，10和20 μmol·L^-1BMS-777607组MCF-7细胞凋亡率降低（P<0.05）。Western blotting法检测，与对照组比较，10、15和20 μmol·L^-1BMS-777607组MCF-7细胞中p-ERK和p-Akt蛋白表达水平明显降低（P<0.05或P<0.01），PARP、CleavedPARP、Bax、CleavedCaspase-3及CleavedCaspase-9蛋白表达水平明显升高（P<0.05或P<0.01）。结论：MST1R抑制剂BMS-777607能够抑制乳腺癌MCF-7细胞增殖并诱导其凋亡，其作用机制与抑制p-ERK和p-Akt表达和促进CleavedPARP、Bax、CleavedCaspase-9及CleavedCaspase-3表达有关。

关键词: BMS-777607, 乳腺肿瘤, 细胞增殖, 细胞凋亡, 巨噬细胞刺激1受体

Abstract: Objective: To investigate the effects of macrophage stimulating 1 receptor (MST1R) inhibitor BMS-777607 on the proliferation and apoptosis of the breast cancer MCF-7 cells, and to elucidate the mechanisms. Methods: The breast cancer MCF-7 cells treated with different concentrations of BMS777607 were divided into control group (0μmol·L^-1 BMS-777607 group) and 0.5,1.0,2.0,5.0,10.0,15.0,and 20.0μmol·L^-1 BMS-777607 groups. MTT method was used to detect the proliferation rates of the MCF-7 cells in various groups, and clone formation assay was used to detect the survival rates of the MCF-7 cells in various groups; EDU imaging and EDU flow cytometry methods were used to detect the proliferation rates of the MCF-7 cells in various groups,and Hoechst33342 staining was used to detect the apoptotic morphology of the MCF-7 cells in various groups; flow cytometry was used to detect the apoptotic rates of the MCF-7 cells in various groups,and Western blotting method was used to detect the expression levels of ERK,p-ERK, Akt,p-Akt, PARP,Cleaved PARP, Bax, Caspase-3,Cleaved Caspase-3,Caspase-9 and Cleaved Caspase-9 proteins in the MCF-7 cells in various groups. Results: The MTT results showed that compared with control group, the proliferation rates of the MCF-7 cells in 5 and 10μmol·L^-1 BMS-777607 groups were increased (P<0.05 or P<0.01). The clone formation assay results showed that compared with control group, the survival rates of the MCF-7 cells in 5 and 20μmol·L^-1 BMS-777607 groups were increased (P<0.05 or P<0.01).The EDU incorporation results showed that compared with control group, the proliferation rates of the MCF-7 cells in 10 and 20μmol·L^-1 BMS-777607 groups were increased (P<0.05 or P<0.01).The Hoechst33342 fluorescence staining results showed that the MCF-7 cells in control group showed the light staining, a small number of MCF-7 cells in 10μmol·L^-1 BMS-777607 group showed the bright nuclei,and most of the MCF-7 cells in 20μmol·L^-1 BMS-777607 group showed the bright nuclei. The flow cytometry results showed that compared with control group, the apoptotic rates of the MCF-7 cells in 10 and 20μmol·L^-1 BMS-777607 groups were decreased (P<0.05). The Western blotting results showed that compared with control group, the expression levels of p-ERK and p-Akt proteins in the MCF-7 cells in 10, 15, and 20μmol·L^-1BMS-777607 groups were significantly decreased(P<0.05 or P<0.01), and the expression levels of PARP, Cleaved PARP, Bax, Cleaved Caspase-3, and Cleaved Caspase-9 proteins were significantly increased(P<0.05 or P<0.01). Conclusion: MST1R inhibitor BMS777607 can inhibit the proliferation of MCF-7 cells and induce the apoptosis, and their mechanisms are related to the inhibition of p-ERK and p-Akt expressions and the promotion of expressions of Cleaved PARP, Bax, Cleaved Caspase-9 and Cleaved Caspase-3.

Key words: BMS-777607, breast neoplasms, cell proliferation, apoptosis, macrophage stimulating 1 receptor

中图分类号:

R737.9

孔雯聪, 贺武斌, 苏荣健, 贾答淇, 谷艳娇, 王月, 杜晓媛. MST1R抑制剂BMS-777607对乳腺癌MCF-7细胞增殖的抑制和凋亡诱导作用[J]. 吉林大学学报(医学版), 2020, 46(03): 451-457.

KONG Wencong, HE Wubin, SU Rongjian, JIA Daqi, GU Yanjiao, WANG Yue, DU Xiaoyuan. Inhibitory effect and apoptosisinduction of MST1R inhibitor BMS-777607 on proliferation of breast cancer MCF-7 cells[J]. Journal of Jilin University(Medicine Edition), 2020, 46(03): 451-457.

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