miR-21通过下调PTEN增强人脑胶质瘤细胞对卡莫司汀耐药  

miR-21 enhances resistance of glioma cells to carmustine by down-re-gulating PTEN

作　　者：胡婷[1] 陈梅香[1] 翁泽平[1] 谢思明[1] 钟雪云[1] 

机构地区：[1]暨南大学医学院病理学系,广东广州510632

出　　处：《中国病理生理杂志》2012年第8期1345-1351,共7页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81072059;No.81101652);广东省高等学校科技创新重点项目(No.CXZD1110)

摘　　要：目的:探讨微小RNA-21(miR-21)在增强人脑胶质瘤细胞对卡莫司汀(BCNU)耐药中的作用及其可能的作用机制。方法:用jetPRIME将miR-21 mimics及阴性对照序列转入人脑胶质瘤细胞SWOZ2,用实时荧光定量PCR法检测BCNU敏感株SWOZ2与BCNU耐药株SWOZ2-BCNU、SWOZ2-miR-21 mimics细胞与对照组细胞中miR-21表达差异,用CCK-8检测这两对细胞对BCNU敏感度的差异,用Western blotting检测这两对细胞中第10号染色体同源缺失性磷酸酶及张力蛋白(PTEN)、磷酸化蛋白激酶B(p-Akt)和P-糖蛋白(P-gp)表达的差异。结果:SWOZ2-BCNU细胞miR-21的表达水平明显高于SWOZ2细胞,转染组SWOZ2-miR-21 mim-ics细胞中miR-21的表达量明显高于对照组;SWOZ2-BCNU细胞BCNU的半数抑制浓度(IC50)明显高于SWOZ2细胞,转染组细胞BCNU的IC50明显高于对照组;与SWOZ2细胞相比,SWOZ2-BCNU细胞中PTEN蛋白的表达明显降低,p-Akt和P-gp蛋白的表达都明显升高;转染后,与对照组相比,转染组细胞中PTEN蛋白的表达明显降低,而p-Akt和P-gp蛋白的表达皆明显升高。结论:miR-21可能通过下调PTEN蛋白表达,增强人脑胶质瘤细胞对BCNU耐药。AIM： To explore the function of miR - 21 in human glioma cells resistant to carmustine[ 1,3 - bis （2 -chloroethyl） -1 -nitrosourea, BCNU ] and to elucidate its related mechanism. METHODS： SWOZ2 cells were transfected with miR -21 mimics （SWOZ2 -miR -21 mimics） or miRNA mimics negative control （control group） by the method of jetPRIME. The real - time fluorescence quantitative PCR was used to detect and compare the levels of miR - 21 expression between BCNU - resistant cell line SWOZ2 - BCNU and BCNU - sensitive cell line SWOZ2, or between SWOZ2 - miR - 21 mimic group and control group. The drug sensitivity of these cells to BCNU was determined by CCK - 8 assay. The protein expression of phosphatase and tensin homology deleted on chromosome 10 （PTEN）, phosphorylated protein ki- nase B （p- Akt） and P- glycoprotein （P- gp） in these cells were also detected by Western blotting. RESULTS： The expression level of miR - 21 was remarkably higher in SWOZ2 - BCNU cells than that in SWOZ2 cells. The expression lev- el of miR-21 was significantly higher in SWOZ2 -miR-21 mimics group than that in control group. The half- maximal inhibitory concentration （ICw ） of BCNU was obviously higher for SWOZ2 - BCNU cells than that for SWOZ2 ceils. The ICso of BCNU was markedly higher in SWOZ2 - miR - 21 mimics group than that in control group. PTEN protein expression was remarkably lower, but p - Akt and P - gp protein expression levels were markedly higher in SWOZ2 - BCNU cells than those in SWOZ2 cells. The protein level of PTEN was significantly lower, but the protein levels of p - Akt or P - gp were distinctly higher in SWOZ2 -miR -21 mimics group than those in control group. CONCLUSION： miR -21 enhances the resistance of human glioma cells to BCNU by down - regulating the expression of PTEN protein.

关 键 词：神经胶质瘤 耐药 MIR-21 

分 类 号：R739.4[医药卫生—肿瘤]