MRPS5对膀胱癌细胞增殖能力及干性特征的影响  

Effect of MRPS5 on proliferation and stemness of bladder cancer cells

作　　者：刘启梁[1] 刘丽梅 刘春刚 钱程 黄庆愿[1] 

机构地区：[1]第三军医大学:高原军事医学系病理生理学与高原病理学教研室,重庆400038 [2]西南医院全军临床病理学研究所,重庆400038

出　　处：《第三军医大学学报》2015年第6期558-562,共5页Journal of Third Military Medical University

摘　　要：目的探讨线粒体核糖体蛋白S5(mitochondrial ribosomal protein S5,MRPS5)对人膀胱癌细胞的增殖能力及其中干细胞干性特征的影响。方法 Western blot检测人膀胱癌及癌旁正常组织MRPS5的表达;构建靶向干扰MRPS5的慢病毒载体,感染膀胱癌细胞株T24,以干扰Scramble序列作为对照,Western blot检测MRPS5干扰效率;细胞活力实验检测细胞增殖能力;流式细胞术检测细胞周期;细胞成球实验观察成球能力;Western blot检测肿瘤干性转录因子Nanog,Oct4,c-Myc和Sox2的表达;小鼠皮下移植瘤实验观察T24体内成瘤能力。结果 MRPS5在膀胱癌组织中的表达高于癌旁正常组织;慢病毒干扰载体PLKO.1-sh MRPS5有效,且干扰MRPS5表达后T24细胞增殖能力下降(P<0.05),周期阻滞于S期,干性因子表达均下调,成球率无变化(P>0.05)但成球直径明显减小;同时小鼠体内成瘤体积减小[(0.784±0.278)vs(0.500±0.245)cm3,P<0.05],瘤质量减轻[(0.862±0.372)vs(0.412±0.248)g,P<0.05]。结论 MRPS5在膀胱癌中较高表达,且干扰MRPS5表达能抑制T24增殖并抑制T24中的干细胞生物学特征。Objective To investigate the effect of mitochondrial ribosomal protein S5 （MRPS5） on proliferation and sternness of human bladder cancer cells. Methods The expression of MRPS5 protein in human bladder cancer tissues and adjacent normal tissues was detected by Western blotting. Lentiviral vector targeting MRPS5 was constructed to infect bladder cancer cell line T24, and lentiviral vector with scramble sequence was used as control. The MRPS5 interference efficiency was analyzed by Western blotting. The cell proliferation was detected by cell viability assay. Cell cycle was measured by flow cytometry. Sphere forming ability was observed by sphere formation experiment. The expression of cancer stem cell related transcription factors （Nanog, Oct4, c-Myc and Sox2 ） was detected by Western blotting. Subcutaneous transplantation tumor experiment of NOD/SCID mice was used to reflect in vivo tumorigenicity. Results The expression of MRPS5 protein in the bladder cancer tissues was higher than that in the adjacent normal tissues. Lentivira] vector PLKO. 1-shMRPS5 was effective for interference to MRPS5 expression. After interference, T24 cell proliferation was decreased （P 〈 0. 05 ）, and the cell cycle was arrested in S phase. Meanwhile, the expression of Nanog, Oct4, c-Myc and Sox2 proteins was all decreased, and the sphere forming ability was not changed （P 〉 0.05）, but the sphere sizes were significantly decreased. In addition, the in vivo tumor volume （0.784 ± 0.278 vs 0. 500 ±0. 245 cm3, P 〈 0. 05 ） and weight （0.862 ± 0. 372 vs 0. 412±0. 248 g, P 〈 0. 05 ） were decreased. Conclusion MRPS5 protein is highly expressed in bladder cancer tissues, and the interference to MRPS5 expression can inhibit the proliferation and sternness of T24 cells.

关 键 词：膀胱癌 线粒体核糖体蛋白S5(MRPS5) 增殖 肿瘤干细胞 

分 类 号：R394.2[医药卫生—医学遗传学] R730.23[医药卫生—基础医学]