中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
Chinese Journal of Experimental Surgery
2015年
10期
2514-2516
,共3页
微小RNA-449a%神经母细胞瘤%增殖%B细胞淋巴瘤/白血病-2
微小RNA-449a%神經母細胞瘤%增殖%B細胞淋巴瘤/白血病-2
미소RNA-449a%신경모세포류%증식%B세포림파류/백혈병-2
MicroRNA-449a%Human neuroblastoma%Proliferation%B cell lymphoma/leukemia-2
目的 检测微小RNA(miRNA,miR)-449a对神经母细胞瘤细胞株SK-N-SH增殖的影响并探讨其作用机制.方法 人工设计合成miR-449a模拟物,构建上调miR-449a表达的细胞株SK-N-SH;实时定量反转录聚合酶链反应(RT-qPCR)检测转染后miR-449a的表达水平;转染48 h后采用细胞计数试剂盒(CCK-8)法检测细胞24、48、72 h的增殖,膜联蛋白V(Annexin-V)/碘化丙锭(PI)法检测细胞凋亡;利用生物信息学方法预测miR-449a的候选靶基因.结果 转染miR-449amimic的细胞中miR-449a的表达水平升高3.5倍;过表达miR-449a后细胞生长速度较对照组细胞明显减慢;过表达miR-449a后细胞凋亡明显增加,细胞总凋亡比例为(21.94±3.37)％,而对照组细胞的总凋亡比例为(11.01±3.51)％,两者差异有统计学意义(P＜0.05);通过靶基因预测软件分析,抗凋亡基因B细胞淋巴瘤/白血病2(bcl-2)是miR-449a的靶基因,过表达miR-449amimic后,内源性bcl-2蛋白的表达水平显著下降.结论 miR-449a可能通过调控bcl-2的转录水平抑制神经母细胞瘤细胞增殖,促进凋亡.
目的 檢測微小RNA(miRNA,miR)-449a對神經母細胞瘤細胞株SK-N-SH增殖的影響併探討其作用機製.方法 人工設計閤成miR-449a模擬物,構建上調miR-449a錶達的細胞株SK-N-SH;實時定量反轉錄聚閤酶鏈反應(RT-qPCR)檢測轉染後miR-449a的錶達水平;轉染48 h後採用細胞計數試劑盒(CCK-8)法檢測細胞24、48、72 h的增殖,膜聯蛋白V(Annexin-V)/碘化丙錠(PI)法檢測細胞凋亡;利用生物信息學方法預測miR-449a的候選靶基因.結果 轉染miR-449amimic的細胞中miR-449a的錶達水平升高3.5倍;過錶達miR-449a後細胞生長速度較對照組細胞明顯減慢;過錶達miR-449a後細胞凋亡明顯增加,細胞總凋亡比例為(21.94±3.37)％,而對照組細胞的總凋亡比例為(11.01±3.51)％,兩者差異有統計學意義(P＜0.05);通過靶基因預測軟件分析,抗凋亡基因B細胞淋巴瘤/白血病2(bcl-2)是miR-449a的靶基因,過錶達miR-449amimic後,內源性bcl-2蛋白的錶達水平顯著下降.結論 miR-449a可能通過調控bcl-2的轉錄水平抑製神經母細胞瘤細胞增殖,促進凋亡.
목적 검측미소RNA(miRNA,miR)-449a대신경모세포류세포주SK-N-SH증식적영향병탐토기작용궤제.방법 인공설계합성miR-449a모의물,구건상조miR-449a표체적세포주SK-N-SH;실시정량반전록취합매련반응(RT-qPCR)검측전염후miR-449a적표체수평;전염48 h후채용세포계수시제합(CCK-8)법검측세포24、48、72 h적증식,막련단백V(Annexin-V)/전화병정(PI)법검측세포조망;이용생물신식학방법예측miR-449a적후선파기인.결과 전염miR-449amimic적세포중miR-449a적표체수평승고3.5배;과표체miR-449a후세포생장속도교대조조세포명현감만;과표체miR-449a후세포조망명현증가,세포총조망비례위(21.94±3.37)％,이대조조세포적총조망비례위(11.01±3.51)％,량자차이유통계학의의(P＜0.05);통과파기인예측연건분석,항조망기인B세포림파류/백혈병2(bcl-2)시miR-449a적파기인,과표체miR-449amimic후,내원성bcl-2단백적표체수평현저하강.결론 miR-449a가능통과조공bcl-2적전록수평억제신경모세포류세포증식,촉진조망.
Objective To investigate the effecof microRN(miRNA, miR)-449on proliferation and apoptosiof neuroblastomSK-N-SH celland the mechanism.MethodSK-N-SH cellwere transfected with mimicontrol and miR-449mimic.The expression level of miR-449wadetermined by real-time quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR).Cell proliferation a24, 48 and 72 h waexamined by cell counting kit-8 (CCK-8) kiafte48 h of transfection.Cell apoptosiwaexamined by annexin-V/ropidium iodide (PI).The putative targetfomiR-449were predicted using bioinformatics.ResultRT-qPCanalysiconfirmed thamiR-449waupregulated 3.5 timein miR-449mimicellthan in mimicontrol cells.miR-449over-expression inhibited cellgrowth a24, 48, and 72 h, respectively acompared with mimicontrol cells.miR-449over-expression could promote cell apoptosiwith apoptosirate being (21.94 ± 3.37)％, which watwofold highethan mimicontrol cell[(11.01 ± 3.51)％ ,P ＜ 0.05].cell lymphoma/leukemia-2 (bcl-2), an antiapoptotimolecule, waidentified to be directly targeof miR-449a.Endogenouprotein expression of bcl-2 wainhibited by miR-449overexpression in SK-N-SH cells.Conclusion miR-449inhibited tumogrowth, and promoted tumoapoptosithrough targeting bcl-2 in human neuroblastoma.
