miR-21抑制物对A549细胞球增殖的影响及机制  

The role and mechanism of miR-21 inhibitor on proliferation of A549 spheres

作　　者：胡欣春[1,2] 魏益平[3] 喻东亮[3] 彭金华[3] 徐建军[3] 

机构地区：[1]南昌大学医学院 [2]江西省胸科医院胸外科 [3]南昌大学第二附属医院心胸外科,江西南昌330006

出　　处：《中国老年学杂志》2014年第9期2456-2459,共4页Chinese Journal of Gerontology

基　　金：国家自然科学基金(No.81160293);国家重大科学仪器设备开发专项(No.2011YQ170067);江西省教育厅青年科学基金(No.GJJ10067)

摘　　要：目的研究miR-21抑制物在A549细胞球增殖的作用及其相关机制。方法在无血清培养基中培养A549细胞球,形成细胞球后进行CD133+CD44+表达检测。实验分为A549细胞球组:未经处理;IHN组:miR-21抑制物转染A549细胞球组;IHN-NC组:miR-21抑制物阴性对照物转染A549细胞球组。Western印迹检测A549细胞和A549细胞球中DKK2、β-catenin蛋白以及QPCR检测miR-21在两种细胞中的表达;合成miR-21抑制物和阴性对照,分别转染A549细胞球48 h后,应用QPCR检测转染前后miR-21的表达变化,以及使用Western印迹分析DKK2、β-catenin蛋白的表达变化,利用噻唑蓝(MTT)检测12、24、48 h细胞的增殖能力。结果流式细胞术检测A549细胞球中CD133+CD44+阳性率为65.800%;IHN组miR-21的表达以及β-catenin蛋白的表达较A549细胞球组和IHN-NC组显著下调(P<0.05),而DKK2蛋白表达显著升高(P<0.05);MTT结果示IHN组和IHN-NC组在12、24、48 h时抑制率显著高于A549 sphere组(P<0.05),IHN组在24、48 h时相点抑制率明显高于IHN-NC组(P<0.05)。结论miR-21抑制物可有效抑制A549 sphere的增殖,其可能成为治疗肺癌的潜在靶点。Objective To study the mechanism and role of miR-21 inhibitors on proliferation of A549 sphere.Methods A549 spheres were cultured in serum-free medium and positive rate of CD 133+CD44+in A549 spheres were detected by FACS .There were A549 sphere, IHN and IHN-NC groups.The expressions of DKK2 andβ-catenin protein and miR-21 were assayed by Western blot and QPCR re-spectively between A549 cells and A549 spheres.miR-21 inhibitiors and negative control were synthetized .A549 spheres were treated with miR-21 inhibitors and negative control respectively for 48 h, and then the expression of miR-21 was detected by Real-time PCR, and the DKK2 andβ-catenin protein expressions were assayed by Western-blot.The inhibition rate of A549 spheres was detected by MTT in 12, 24, 48 h timepoint.Results CD133+CD44+A549 cells were sorted.Positive rate of CD133+CD44+was 65.800%in A549 spheres.Compared with those of A549 sphere and IHN-NC groups , the expressions of miR-21 and β-catenin protein in IHN group were significantly decreased and DKK2 protein expression was higher (P<0.05).Compared with that of A549 sphere group, the proliferation inhibition rate of IHN and IHN-NC group in 12, 24, 48 h were higher (P<0.05 respectively), and the proliferation inhibition rate of IHN was significant higher than that of IHN-NC group(P<0.05).Conclusions miR-21 inhibitors could effectively inhibit the A549 sphere proliferation, and it might be-come a novel target for treatment non small cell lung cancer .

关 键 词：MIR-21 抑制物 肺癌 DKK2 

分 类 号：R73[医药卫生—肿瘤]