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浙江大学学报（医学版）
浙江大学学报（医学版）  2017, Vol. 46 Issue (4): 364-370    DOI: 10.3785/j.issn.1008-9292.2017.08.04
乳腺癌分子医学专题     
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
李钰1, 王月月1, 王海凤1, 张凌宇1, 丁勇兴2, 陈素莲3, 杨清玲3, 陈昌杰3
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
Effects of lncRNA RP11-770J1.3 and TMEM25 expression on paclitaxel resistance in human breast cancer cells
LI Yu1, WANG Yueyue1, WANG Haifeng1, ZHANG Lingyu1, DING Yongxing2, CHEN Sulian3, YANG Qingling3, CHEN Changjie3
1. Clinical Testing and Diagnose Experimental Center, Bengbu Medical College, Bengbu 233000, China;
2. Department of Oncology, the Third People's Hospital of Bengbu, Bengbu 233000, China;
3. Department of Biochemistry & Molecular Biology, Bengbu Medical College, Bengbu 233000, China
 全文: PDF(2245 KB)  
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。
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    
Abstract:

Objective:To investigate the effects of long non-coding RNA(lncRNA) 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) cells were detected by quantitative RT-PCR. The synthetic interfering fragments of lncRNA RP11-770J1.3 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 multidrug-resistance genes and proteins were detected by qRT-PCR and Western blot, respectively. Results:lncRNA 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 lncRNA 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 lncRNA RP11-770J1.3 and TMEM25 were both down-regulated (all P<0.05). Conclusion:lncRNA RP11-770J1.3 and TMEM25 are highly expressed in MCF-7/PR cells, and the down-regulation of lncRNA RP11-770J1.3 and TMEM25 can enhance paclitaxel sensitivity in MCF-7/PR cells.
Key words: RNA    Membrane proteins/metabolism    Breast neoplasms/physiopathology    Polymerase chain reaction    Paclitaxel/pharmacology    Cell line, tumor/pathology    Tumor cells, cultured    Drug resistance, neoplasm
收稿日期: 2017-05-05 出版日期: 2017-08-25
CLC:  R737.9  
基金资助:

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）
通讯作者: 陈昌杰(1968-),男,博士,教授,硕士生导师,主要从事肿瘤分子生物学研究;E-mail:tochenchangjie@163.com;http://orcid.org/0000-0001-9646-320X     E-mail: tochenchangjie@163.com
作者简介: 李钰(1991-),女,硕士研究生,主要从事感染性疾病分子生物学机制研究;E-mail:liyu1234july@163.com;http://orcid.org/0000-0002-8465-6183
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引用本文:

李钰 等. 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[J]. 浙江大学学报（医学版）, 2017, 46(4): 364-370.

LI Yu, WANG Yueyue, WANG Haifeng, ZHANG Lingyu, DING Yongxing, CHEN Sulian, YANG Qingling, CHEN Changjie. Effects of lncRNA RP11-770J1.3 and TMEM25 expression on paclitaxel resistance in human breast cancer cells. Journal of ZheJiang University(Medical Science), 2017, 46(4): 364-370.

链接本文:

http://www.zjujournals.com/xueshu/med/CN/10.3785/j.issn.1008-9292.2017.08.04    或    http://www.zjujournals.com/xueshu/med/CN/Y2017/V46/I4/364

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