microRNA-429对人肺腺癌细胞SPC-A1增殖抑制的影响  

Effects of miR-429 on the inhibits of cell proliferation in the human lung adenocarcinoma cell line SPC-A1

作　　者：乐涵波[1] 竺王玉[1] 刘晓光[1] 何剑营[1] 陈冬冬[1] 续力云[1] 张永奎[1] 

机构地区：[1]温州医科大学附属舟山医院心胸外科肺癌研究中心,浙江316004

出　　处：《中华胸心血管外科杂志》2014年第12期733-735,共3页Chinese Journal of Thoracic and Cardiovascular Surgery

基　　金：舟山市科技局重大项目（2011C12039,10137）

摘　　要：目的 探索microRNA-429（miR-429）对人肺腺癌细胞SPC-A1增殖抑制的影响.方法 合成miR-429前体（Pre-miRTM miR-429 precursor）,通过脂质体转染至SPC-A1细胞,荧光定量PCR法检测miR-429表达水平,CCK-8法检测细胞活性,Annexin V Assay检测细胞凋亡情况,流式细胞术检测细胞周期情况,Western-blot检测细胞周期蛋白情况.结果 pre-miR-429转染后,SPC-A1细胞miR-429表达较对照组明显增高,差异有统计学意义（P＜0.001）;CCK-8检测结果显示,转染pre-miR-429 48、72 h后,细胞的增殖活性较空白组和对照组明显下降,差异有统计学意义（P=0.0167、0.0383,P=0.0320、0.0465）;Annexin V Assay显示pre-miR-429转染24h后,pre-miR-429组与对照组比较凋亡率无明显区别,差异无统计学意义（P＞0.05）;细胞周期结果显示pre-miR-429组G0/G1期细胞比例降低,S期细胞比例增高,细胞周期阻滞于S期,差异有统计学意义（P =0.0010,0.0006;P=0.0068,0.0133）;pre-miR-429组Cyc-lin E蛋白表达与对照组比较无明显区别.结论 在肺腺癌细胞SPC-A1中增加miR-429表达量,可抑制肺癌细胞增殖,促进细胞周期阻滞.miR-429在肺腺癌SPC-A1细胞中可能发挥潜在的抑癌作用.Objective To assess the impact of miR-429 on lung adenocarcinoma cell SPC-A1 growth inhibition.Methods Pre-miRTM miR-429 precursor was synthesized and transfected to the SPC-A1 cells by liposome; qRT-PCR assay was used to quantify the miR-429 expression levels; The proliferation of SPC-A1 cells was evaluated by Cell Counting Kit-8 （CCK8）.The cell apoptosis was evaluated by Annexin V Assay; The cell cycles of each group were assayed by flow cytometry;Western-blot was used to analyze the expression of cylines.Results The expression level of miR-429 was highly induced after transfection （P ＜ 0.001） ;CCK-8 assay showed the cell proliferation activity of pre-miR-429 group was lower than that of blank and control group 48h and 72 h after transfection（P =0.0167,0.0383,P =0.0320,0.0465）,whereas the apoptosis rate had no significant difference between pre-miR-429 and control 24h after transfection by Annexin V Assay（P ＞ 0.05） ; The flow cytometry at 48h after transfection showed that miR-429 decreased the percentage of cells in G1 phase,but increased in S phase,indicating the cell cycle arrest at S phase（P =0.0010,0.0010 ; P =0.0068,0.0133） ; however,the expression level of Cyclin E in pre-miR-429 group had no difference compared with control.Conclusion miR-429 could inhibit cell proliferation and promote cell cycle arrest of lung adenocarcinoma cell SPC-A1.miR-429 may play a potential tumor suppressor role in lung adenocarcinoma cell SPC-A1.

关 键 词：肺肿瘤 腺癌 miR-429 增殖 细胞周期 

分 类 号：R734.2[医药卫生—肿瘤]