miRNA-199a-5p对骨肉瘤细胞增殖及自噬的影响  

Effect of miRNA-199a-5p on Cell Proliferation and Autophagy in Osteosarcoma

作　　者：刘念[1] 张红[2] 薛中柱[3] 

机构地区：[1]郑州大学附属南阳市中心医院骨二科,南阳473400 [2]郑州大学附属南阳市中心医院肾病风湿免疫科,南阳473400 [3]郑州大学附属南阳市中心医院医务科,南阳473400

出　　处：《肿瘤防治研究》2017年第3期197-201,共5页Cancer Research on Prevention and Treatment

摘　　要：目的探讨mi RNA-199a-5p对人骨肉瘤细胞株U2OS增殖及自噬的影响及其可能的作用机制。方法实时定量PCR法检测32例骨肉瘤患者的病理组织标本及配对癌旁正常组织中mi RNA-199a-5p的表达水平。将mi RNA-199a-5p mimics转染入U2OS细胞,使其过表达mi RNA-199a-5p,采用CCK-8法和流式细胞仪检测细胞增殖及周期的变化;Western blot检测周期相关蛋白(cyclin D1、P27、p Rb)及自噬相关蛋白(Beclin 1、LC3Ⅱ/Ⅰ)表达水平。结果 mi RNA-199a-5p在骨肉瘤组织及癌细胞U2OS中低表达,且表达水平与肿块大小、远距转移、Enneking临床分期及病理分级密切相关。过表达mi RNA-199a-5p后,U2OS细胞的增殖能力显著降低(P<0.05),同时cyclin D1、p Rb表达水平显著下降,而P27、LC3Ⅱ/Ⅰ、Beclin 1的表达水平显著增加(P<0.05)。结论 mi RNA-199a-5p能够抑制人骨肉瘤细胞株U2OS的增殖并激活细胞自噬;其作用机制可能与下调cyclin D1、上调P27、抑制p Rb蛋白的磷酸化、造成细胞周期G1/S阻滞有关。提示mi RNA-199a-5p可以作为骨肉瘤临床治疗的潜在作用位点。Objective To investigate the effect of mi RNA-199a-5p on cell proliferation and autophagy in osteosarcoma cell line U2 OS and its possible mechanism.Methods The expression of mi RNA-199a-5p were detected by q RT-PCR after transfection with the mi RNA-199a-5p mimics in U2 OS cells in paraffin section and paired adjacent normal tissue from 32 patients with osteosarcoma.The proliferation were measured by CCK-8 assay and flow cytometry assay.The expression of cell cycle related protein（cyclin D1,P27,p Rb） and autography related protein（Beclin 1,LC3Ⅱ/Ⅰ） were measured by Western blot assay.Results Mi RNA-199a-5p was lowly expressed in both osteosarcoma tissues and U2 OS cells（P〈0.05）.The expression level of mi RNA-199a-5p was correlated with the tumor size,distant metastasis,Enneking stage and pathological grade stage（P〈0.05）.Overexpression of mi RNA-199a-5p suppressed U2 OS cell proliferation（P〈0.05）,and downregulated the expression of cyclin D1 and p Rb,while upregulated the expression of P27,Beclin 1 and LC3Ⅱ/Ⅰ（P〈0.05）.Conclusion mi RNA-199a-5p suppresses cell proliferation but induces cell autophagy.And its mechanism may be related to thedownregulation of cyclin D1 and upregulation of P27 protein expression,then supressed the phosphorylation of p Rb,resulting in G1/S arrest in human osteosarcoma cell line U2 OS.It suggests that mi RNA-199a-5p may be used as a potential new target for targeted therapy of osteosarcoma.

关 键 词：miRNA-199a-5p 骨肉瘤 增殖 周期 自噬 

分 类 号：R738[医药卫生—肿瘤]