BMI-1抑制剂PTC-596对人乳腺癌MCF-7细胞增殖、周期和凋亡的影响  

Effect of BMI-1 inhibitor PTC-596 on proliferation,cell cycle,and apoptosis of human breast cancer MCF-7cells

作　　者：李玉玲 杨建林[1] 崔芝 王静[1] 吕亚丰 曹春雨[1] LI Yuling;YANG Jianlin;CUI Zhi;WANG Jing;LV Yafeng;CAO Chunyu(College of Basic Medi-cal Sciences,China Three Gorges University,Hubei Key Laboratory of Tumor Microenvironment and Immuno-therapy,Yichang 443002,China)

机构地区：[1]三峡大学基础医学院,肿瘤微环境与免疫治疗湖北省重点实验室,湖北宜昌443002

出　　处：《实用医学杂志》2023年第18期2317-2322,2329,共7页The Journal of Practical Medicine

基　　金：国家自然科学基金资助项目(编号:81372265,30772590);湖北省卫生健康科研基金(编号:WJ2019H532);肿瘤微环境与免疫治疗湖北省重点实验室开放基金项目(编号:2019KZL01,2016KZL04)。

摘　　要：目的 分析BMI-1抑制剂PTC-596对人乳腺癌MCF-7细胞增殖、周期和凋亡的影响。方法以人乳腺癌MCF-7细胞为肿瘤细胞模型,正常培养为阴性对照组,25、50 nmol/L PTC-596处理SGC-7901细胞48 h为低、高药物浓度实验组。利用CCK8法分析细胞增殖能力;流式细胞术分别结合PI单染、DCFHDA探针、JC-1探针以及PI/FITC-Annexin V双染分析细胞周期、活性氧(reactive oxygen species, ROS)累积、线粒体膜电位和凋亡细胞比例;Western blot法检测细胞BIM-1蛋白和周期相关蛋白CyclinD1、CyclinB1、P21以及凋亡相关蛋白Bax、Bcl-2、c-PARP蛋白相对表达水平。结果 与对照组相比,PTC-596高效抑制人乳腺癌MCF-7细胞的增殖,处理48 h后IC_(50)为(49.33±7.02)nmol/L。低、高药物浓度实验组细胞中BMI-1表达显著减少(P <0.01)。实验组细胞中CyclinB1和P21相对表达增加,CyclinD1表达减少,细胞有丝分裂被抑制,出现G_2/M期周期阻滞;同时活性氧累积增多,线粒体膜电位下降,Bax表达上调,Bcl-2表达下调,c-PARP增加,凋亡细胞比例从2.04%显著上升为10.56%、26.74%。与对照组相比较,以上结果差异均有统计学意义(P <0.05)。结论 PTC-596高效杀伤人乳腺癌MCF-7细胞,其机制可能与抑制BMI-1、诱导细胞周期阻滞和内源性线粒体途径细胞凋亡密切相关。Objective To investigate the effect of a novel BMI-1 inhibitor PTC-596 on the proliferation,cell cycle,and apoptosis of human breast cancer MCF-7 cells.Methods MCF-7 cells treated with 0 nmol/L PTC-596 were used as control group;MCF-7 cells treated with 25,and 50 nmol/L PTC-596 as experimental group.The CCK8 assay was used to evaluate the celular proliferation ability;DCFH-DA probe/flow cytometry(FCM)was performed to detect the change of cellular reactive oxygen species(ROS).PI single-staining/FCM was used to analyze cell cycle;PI/FITC-Annexin V/FCM to analyze the proportion of apoptotic cells.JC-1/FCM was performed to detect the change of mitochondrial membrane potential(MMP);western blot to analyze the expression of BMI-1,cell cycle related proteion(CyclinD1,CyclinB1,and P21)and apoptosis related proteion(Bax,Bcl-2,and c-PARP).Results PTC-596 could significantly inhibit MCF-7 cell proliferation,with the IC50 value of 49.33±7.02 nmol/L(24 h).BMI-1 expression was significantly decreased in the low-and high-drug concentration experi-mental group(P<0.01).The relative expression of CyclinB1 and P21 in the experimental groups were increased(P<0.01),while the expression of CyclinD1 was decreased(P<0.01),and cell mitosis was inhibited(P<0.01),resulting in G2/M cycle arrest(P<0.01).At the same time,the accumulation of reactive oxygen species was in-creased(P<0.01);mitochondrial membrane potential was decreased(P<0.01).Bax expression was up-regulated(P<0.01),while Bcl-2 expression was down-regulated(P<0.01);c-PARP was increased(P<0.01),and the proportion of apoptotic cells was significantly increased from 2.04%to 10.56%and 26.74%(P<0.01).Conclusion PTC-596 can effectively inhibit the proliferation of human breast cancer MCF-7cells,and its mechanism may be closely related to the inhibition of BMI-1,the induction of cell cycle arrest,and the endogenous mitochondrial apoptosis.

关 键 词：原癌基因BMI-1 PTC-596 人乳腺癌MCF-7细胞 细胞增殖 细胞周期 凋亡 

分 类 号：R730.53[医药卫生—肿瘤]