转染miRNA27a抑制物对卵巢癌耐药细胞系顺铂敏感性的研究

DOWN-REGULATE MIRNA27A AND ITS RELATIONSHIP WITH DRUG RESISTANCE IN HUMAN OVARIAN CANCER SKOV3/DDP

目的研究下调miRNA27a对卵巢癌细胞系skov3/DDP顺铂耐药性的影响。方法培养卵巢癌细胞系skov3,使用顺铂诱导,建立卵巢癌顺铂耐药细胞系skov3/DD,MTT法检测细胞耐药系数,Real-time RT-PCR检测miRNA27a在skov3/DDP及skov3中的表达情况,将miRNA27a抑制物转入skov3/DDP,Real-time RT-PCR检测MDR1基因的转录水平,流式细胞术检测细胞凋亡,Western Blot检测P-gp蛋白表达水平。结果与skov3相比,skov3/DDP耐药系数为5.08;miRNA27a在skov3/DDP中的表达明显上升;转染miRNA27a抑制物之后,与阴性对照相比,skov3/DDP中的MDR1的转录水平无明显差异(P>0.05);但与SKOV3相比,SKOV3/DDP中p-gp蛋白的表达水平明显升高,约为SKOV3的1.5倍(P<0.05),而转染了miRNA27a抑制物之后,SKOV3/DDP中P-gp的蛋白表达水平明显降低,与SKOV3相比,表达量减少了32.7%(P<0.05)。结论 miRNA27a在skov3/DDP中高表达,导致了skov3/DDP对顺铂产生了明显的耐药性,其机制主要是通过在P-gp的翻译水平诱导其耐药性的产生。

Objective To research the effect of cisplatin resistance with down-regulation miRNA27 ain ovarian cancer cell line skov3/DDP.Methods Cultivate the ovarian cancer cell line skov3,establish cisplatin resistant ovarian cancer cell line which induced by cisplatin.Detect the cell resistance coefficient by MTT,Detect the level of miRNA27 ain skov3cell and skov3/DDP cell by Real-time RT-PCR,then transfection the inhibitor of miRNA27 a,Real-time RT-PCR detect the Transcription level of MDR-1,detect the translation level of P-gp.Results Compared with skov3,the resistance index of skov3/DDP was 5.08;the level of miRNA27 ain skov3/DDP was significantly up-regulation.Compared with negative control,the Transcription level of MDR-1in skov3/DDP was not significant difference（P〉0.05）;but compared with SKOV3,the level of P-gp was significantly up-regulation（P〈0.05）,and the level of P-gp was down-regulation by transfection the inhibitor of miRNA27a（P〈0.05）.Conclusion The drug resistance of cisplatin in skov3/DDP was induced by over expressed of miRNA27 a,the main mechanism was up-regulation the translational level of P-gp.