长链非编码RNA RP11-770J1.3和跨膜蛋白25对紫杉醇耐药人乳腺癌细胞株耐药性的影响  

Effects of IncRNA RP11-770J1.3 and TMEM25 expression on paclitaxel resistance in human breast cancer cells

作　　者：李钰[1] 王月月 王海凤[1] 张凌宇[1] 丁勇兴[2] 陈素莲[3] 杨清玲[3] 陈昌杰[3] 

机构地区：[1]蚌埠医学院临床检验诊断学实验中心,安徽蚌埠233000 [2]蚌埠市第三人民医院普外肿瘤科,安徽蚌埠233000 [3]蚌埠医学院生物化学与分子生物学教研室,安徽蚌埠233000

出　　处：《浙江大学学报（医学版）》2017年第4期364-370,共7页Journal of Zhejiang University(Medical Sciences)

基　　金：安徽省教育厅自然科学重大项目(KJ2015ZD29,KJ2016SD37);安徽省自然科学基金(1508085MH159);安徽省高校学科(专业)拔尖人才学术资助重点项目(gxbjZD2016069);安徽省蚌埠市科技计划项目(20150309);蚌埠医学院研究生创新项目(Byycxz1607,Byycx1607,Byycx1615)

摘　　要：目的:探讨长链非编码RNA(lncRNA)RP11-770J1.3和跨膜蛋白25(TMEM25)对紫杉醇耐药人乳腺癌细胞株耐药性的影响。方法:利用实时定量PCR检测lncRNA RP11-770J1.3和TMEM25在人乳腺癌细胞MCF-7和紫杉醇耐药细胞株(MCF-7/PR)中的表达。在MCF-7/PR中转染lncRNA RP11-770J1.3和TMEM25的干扰片段,磺酰罗丹明B法检测MCF-7/PR对紫杉醇药物敏感性的变化,并运用实时定量PCR和蛋白质印迹法检测多药耐药相关蛋白(MRP)、乳腺癌耐药蛋白(BCRP)、多药耐药基因1(MDR1)及其编码产物P-gp mRNA和蛋白水平的改变。结果:lncRNA RP11-770J1.3和TMEM25在MCF-7/PR中表达上调(P<0.05或P<0.01)。与空白对照组和紫杉醇阴性对照组比较,干扰lncRNA RP11-770J1.3和TMEM25的表达可以提高MCF-7/PR对紫杉醇的药物敏感性,并下调耐药相关基因MRP、BCRP、MDR1及其编码产物P-gp的表达(均P<0.05);与单独干扰lncRNA RP11-770J1.3或TMEM25比较,同时干扰lncRNA RP11-770J1.3和TMEM25的作用更加明显(P<0.05)。结论:MCF-7/PR中lncRNA RP11-770J1.3和TMEM25的表达上调,联合干扰lncRNA RP11-770J1.3和TMEM25的表达可以提高MCF-7/PR对紫杉醇的敏感性。Objective： To investigate the effects of long non-coding RNA（IncRNA） RP11-770J1.3 and transmembrane protein 25 （TMEM25） on paclitaxel resistance in human breast cancer MCF-7/PR cell line. Methods： The expression of lncRNA RP11- 770J1.3 and TMEM25 in human breast cancer MCF-7 （paclitaxel sensitive） and MCF- 7/PR（ paclitaxel resistant） ceils were detected by quantitative RT-PCR. The synthetic interfering fragments of lncRNA RPll-770J1. 3 and TMEM25 were transfected into MCF-7/PR cells. Sulforhodamine B assay was used to detect the sensitivity of MCF-7/PR cells to paclitaxel after interference of lncRNA RP11-770J1.3 and TMEM25. The expression of muhidrug-resistance genes and proteins were detected by qRT-PCR and Western blot, respectively. Results： IncRNA RP11-770J1.3 and TMEM25 were highly expressed in MCF-7/PR cells, and were significantly down-regulated after transfection of synthetic interfering fragments. Down-regulation of IncRNA RP11-770J1. 3 and TMEM25 enhanced the sensitivity of MCF-7/PR cells to paclitaxel, and inhibited the expression of MRP, BCRP and MDR1/P-gp （ all P 〈 0.05 ）. Such effects were more significant when IncRNA RP11-770J1.3 and TMEM25 were both down-regulated （ all P 〈 O. 05）. Conclusion： lncRNA RPll-770J1. 3 and TMEM25 are highly expressed in MCF-7/PR ceils, and the down-regulation of lncRNA RP11-770J1.3 and TMEM25 can enhance paclitaxel sensitivity in MCF-7/PR cells.

关 键 词：RNA 膜蛋白质类/代谢 乳腺肿瘤/病理生理 聚合酶链反应 紫杉 酚/药理学 细胞系 肿瘤/病理学 肿瘤细胞 培养的 抗药性 肿瘤 

分 类 号：R737.9[医药卫生—肿瘤]