CAJ | 학술논문

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.
목적 탐색microRNA-429(miR-429)대인폐선암세포SPC-A1증식억제적영향.방법 합성miR-429전체(Pre-miRTM miR-429 precursor),통과지질체전염지SPC-A1세포,형광정량PCR법검측miR-429표체수평,CCK-8법검측세포활성,Annexin V Assay검측세포조망정황,류식세포술검측세포주기정황,Western-blot검측세포주기단백정황.결과 pre-miR-429전염후,SPC-A1세포miR-429표체교대조조명현증고,차이유통계학의의(P＜0.001);CCK-8검측결과현시,전염pre-miR-429 48、72 h후,세포적증식활성교공백조화대조조명현하강,차이유통계학의의(P=0.0167、0.0383,P=0.0320、0.0465);Annexin V Assay현시pre-miR-429전염24h후,pre-miR-429조여대조조비교조망솔무명현구별,차이무통계학의의(P＞0.05);세포주기결과현시pre-miR-429조G0/G1기세포비례강저,S기세포비례증고,세포주기조체우S기,차이유통계학의의(P =0.0010,0.0006;P=0.0068,0.0133);pre-miR-429조Cyc-lin E단백표체여대조조비교무명현구별.결론 재폐선암세포SPC-A1중증가miR-429표체량,가억제폐암세포증식,촉진세포주기조체.miR-429재폐선암SPC-A1세포중가능발휘잠재적억암작용.
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.