微小RNA-25-3p靶向下调甘油磷酸二酯酶磷酸结构域5基因对乳腺癌细胞顺铂耐药性的影响  

Effect of miR-25-3p targeted down-regulation of glycerophosphodiester phosphodiesterase domain 5 gene on cisplatin resistance of breast cancer cells

作　　者：缪玖麟 王建荣[1] 徐志敏 邱卫明 陈保华[1] 李新建[1] MIAO Jiulin;WANG Jianrong;XU Zhimin;QIU Weiming;CHEN Baohua;LI Xinjian(General Surgery Department,Yingtan Medical District,No.908 Hospital of Chinese People's Liberation Army Joint Support Force,Yingtan,Jiangxi 335000,China)

机构地区：[1]中国人民解放军联勤保障部队第九〇八医院鹰潭医疗区普通外科,江西鹰潭335000

出　　处：《安徽医药》2020年第10期1937-1942,共6页Anhui Medical and Pharmaceutical Journal

摘　　要：目的探讨微小RNA-25-3p(miR-25-3p)靶向甘油磷酸二酯酶磷酸结构域5(GDPD5)对乳腺癌细胞顺铂耐药性的影响及分子机制。方法本研究起止时间为2018年10月至2019年6月,人乳腺癌细胞MCF-7和乳腺癌顺铂耐药细胞MCF-7/DDP购于中国科学院上海细胞研究所,采用实时荧光定量PCR(qRT-PCR)和蛋白质印迹法(Western blot)检测正常乳腺癌细胞MCF-7和乳腺癌顺铂耐药细胞MCF-7/DDP中miR-25-3p、GDPD5和谷胱甘肽巯基转移酶π(GST-π)的表达水平。四甲基偶氮唑盐比色(MTT)法检测过表达miR-25-3p或过表达miR-25-3p联合顺铂对MCF-7/DDP细胞的增殖抑制作用,蛋白质印迹法检测GDPD5、GST-π和细胞周期蛋白D1(Cyclin D1)蛋白的表达水平。双荧光素酶报告基因实验和蛋白质印迹法验证miR-25-3p和GDPD5的靶向关系。结果与MCF-7细胞相比,MCF-7/DDP细胞中miR-25-3p的表达显著下调,GDPD5和GST-π的表达显著上调。过表达miR-25-3p后,MCF-7/DDP细胞存活率显著降低[(50.36±5.04)%比(100.00±10.11)%],CyclinD1相对表达水平降低(0.40±0.05)比(1.12±0.11),GST-π表达水平降低(0.35±0.04)比(1.15±0.12);且过表达miR-25-3p可降低MCF-7/DDP细胞的顺铂耐药性。miR-25-3p可靶向负性调控GDPD5的表达。过表达GDPD5可部分逆转miR-25-3p对MCF-7/DDP细胞增殖和顺铂耐药性的影响。结论miR-25-3p通过靶向下调GDPD5抑制MCF-7/DDP细胞存活,进而降低MCF-7/DDP细胞对顺铂的耐药性。Objective To investigate the effect of microRNA-25-3 p(miR-25-3 p)targeting glycerophosphodiester phosphodiesterase domain 5(GDPD5)on cisplatin resistance in breast cancer cells and its molecular mechanism.MethodsThe study was conducted from October 2018 to June 2019.Human breast cancer cells MCF-7 and cisplatin-resistant breast cancer cells MCF-7/DDP were purchased from Shanghai Institute of Cell Research,Chinese Academy of Sciences.Real-time fluorescent quantitative PCR(q RT-PCR)and Western blot were used to detect the expression level of miR-25-3 p,GDPD5 and glutathione transferase(GST-π)in breast cancer cells MCF-7 and cisplatin-resistant breast cancer MCF-7/DDP cells.Methyl thiazolyl tetrazolium assay(MTT)assay was used to detect the inhibitory effect of over-expressing miR-25-3 p or over-expressing miR-25-3 p combined with cisplatin on the proliferation of MCF-7/DDP cells,and Western blot was used to detect the expression of GDPD5,GST-πand CyclinD1 proteins.The dual luciferase reporter gene assay and Western blot were used to verify the targeting relationship between miR-25-3 p and GDPD5.ResultsCompared with MCF-7 cells,the expression of miR-25-3 p in MCF-7/DDP cells was significantly down-regulated,while the expression of GDPD5 and GST-πwas significantly up-regulated.After overexpression of miR-25-3 p,the survival rate of MCF-7/DDP cells was significantly reduced[(50.36±5.04)%vs.(100.00±10.11)%],and the relative expression of CyclinD1 was decreased(0.40±0.05)vs.(1.12±0.11);the expression of GST-πwasdecreased(0.35±0.04)vs.(1.15±0.12);and overexpression of miR-25-3 p could reduce the cisplatin resistance of MCF-7/DDP cells.miR-25-3 p negatively regulated the expression of GDPD5.Over-expression of GDPD5 partially reversed the effect of miR-25-3 p on the proliferation and cisplatin resistance of MCF-7/DDP cells.ConclusionmiR-25-3 p inhibits the survival of MCF-7/DDP cells by down-regulating GDPD5,thereby reducing the resistance of MCF-7/DDP cells to cisplatin.

关 键 词：乳腺肿瘤 抗药性 肿瘤 miR-25-3p 甘油磷酸二酯酶磷酸结构域5 顺铂 谷胱甘肽转移酶 

分 类 号：R737.9[医药卫生—肿瘤]