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Journal of ZheJiang University(Medical Science)  2017, Vol. 46 Issue (4): 364-370    DOI: 10.3785/j.issn.1008-9292.2017.08.04
    
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
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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 wordsRNA      Membrane proteins/metabolism      Breast neoplasms/physiopathology      Polymerase chain reaction      Paclitaxel/pharmacology      Cell line, tumor/pathology      Tumor cells, cultured      Drug resistance, neoplasm     
Received: 05 May 2017      Published: 25 August 2017
CLC:  R737.9  
Cite this article:

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.

URL:

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


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

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 

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