Mus81基因沉默对MCF-7人乳腺癌细胞系增殖凋亡及化疗敏感性的影响  

Effects of methyl methansulfonate and UV sensitive gene clone 81 on proliferation,apoptosis and chemosensitivity of MCF-7 human breast cancer cell line

作　　者：常颖智 赵俐 张年伟 熊白玉 吴帆 CHANG Yingzhi;ZHAO Li;ZHANG Nianwei;XIONG Baiyu;WU Fan(Department of Breast Surgery,Guangzhou First People′s Hospital,the Second Affiliated Hospital of South China University of Technology,Guangzhou 510180,China;不详)

机构地区：[1]广州市第一人民医院,华南理工大学附属第二医院乳腺外科,广州510180 [2]广州市红十字会医院普通外科,广州510220

出　　处：《实用医学杂志》2022年第12期1475-1479,1485,共6页The Journal of Practical Medicine

基　　金：国家自然科学基金面上项目(编号:81974442);广东省自然科学基金面上项目(编号:2020A1515010799);广州市医药卫生科技项目(编号:20131A011006)。

摘　　要：目的研究Mus81基因沉默对MCF-7人乳腺癌细胞系增殖、凋亡和化疗敏感性的影响并初步探讨其调控机制。方法首先构建Mus81沉默的MCF-7乳腺癌细胞系,使用噻唑蓝(MTT)法和平板克隆实验检测细胞生长和增殖,碘化丙锭(PI)染色法和膜联蛋白V(Annexin V)-APC染色法检测细胞周期及凋亡的变化。qRT-PCR检测Mus81沉默后下游基因的表达,MTT法检测化疗药物的50%抑制浓度(IC50)及逆转指数(RI)。结果(1)MTT生长曲线和平板克隆实验显示Mus81基因沉默后MCF-7细胞系增殖显著减缓(P<0.001),细胞克隆数明显下降(P<0.001)。(2)Annexin V-APC和PI染色检测显示MCF-7细胞凋亡率升高(P<0.001),并出现明显的G2/M期阻滞。(3)MTT检测结果显示,Mus81沉默后MCF-7细胞对顺铂、阿霉素、表阿霉素和5-氟尿嘧啶的IC50值明显降低,RI依次为5.118、3.070、3.027及8.997。(4)qRT-PCR及Western blot检测下游信号通路表达的结果显示,Mus81沉默后MCF-7细胞中的APAF1、APC和PTEN基因表达升高,MAPK3和MAPK1基因表达降低。结论Mus81基因沉默可明显抑制MCF-7人乳腺癌细胞系的生长增殖并诱导G2/M期阻滞及细胞凋亡,还可提高MCF-7细胞对顺铂、阿霉素、表阿霉素和5-氟尿嘧啶的敏感性,其机制可能与调控APAF1、APC、PTEN、MAPK1及MAPK3等下游基因的表达有关,提示Mus81可能是一个潜在的乳腺癌治疗靶点。Objective To study the effect of Mus81 gene silencing on the proliferation,apoptosis and chemosensitivity of MCF-7 human breast cancer cell line,and to preliminarily explore its regulatory mechanism.Methods Mus81-silenced MCF-7 breast cancer cell line was initially constructed.Cell growth and proliferation were detected by thiazolyl blue(MTT)method and plate cloning assay.Propidium iodide(PI)staining and Annexin V-APC staining were used to detect changes in cell cycle and apoptosis.qRT-PCR detected the expression of downstream genes after Mus81 was silenced,and MTT method detected the 50%inhibitory concentration(IC50)and reversal index(RI)of chemotherapy drugs.Results The MTT growth curve and plate cloning assay showed that the proliferation of MCF-7 cell line was significantly retarded after Mus81 gene silencing(P<0.001),and the number of cell clones was significantly decreased(P<0.001).Annexin V-APC and PI staining showed that the apoptotic rate of MCF-7 cells increased(P<0.001),and obvious G2/M phase arrest developed.MTT test results showed that the IC50values of MCF-7 cells to cisplatin,doxorubicin,epirubicin and 5-fluorouracil were significantly reduced after Mus81 silencing,and the RI was 5.118,3.070,3.027 and 8.997 respectively.The results of qRT-PCR and Western blot for detecting the expression of downstream signaling pathways showed that the expressions of APAF1,APC and PTEN genes in MCF-7 cells increased after Mus81 silencing,while the expressions of MAPK3 and MAPK1 genes decreased.Conclusions Mus81 gene silencing can significantly inhibit the growth and proliferation of the MCF-7 human breast cancer cell line and induce G2/M phase arrest and apoptosis.It can also increase the sensitivity of MCF-7 cells to cisplatin,doxorubicin,epirubicin and 5-fluorouracil.The mechanism may be related to the regulation of the expressions of downstream genes APAF1,APC,PTEN,MAPK1 and MAPK3,suggesting that Mus81 may be a potential therapeutic target of breast cancer.

关 键 词：甲磺酸盐及紫外线敏感性81号基因 乳腺癌 增殖 凋亡 化疗敏感性 

分 类 号：R737.9[医药卫生—肿瘤]