中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
Chinese Journal of Experimental Surgery
2015年
9期
2196-2198
,共3页
微小RNA-10b%胰腺癌%增殖
微小RNA-10b%胰腺癌%增殖
미소RNA-10b%이선암%증식
MicroRNA-10b%Pancreatic cancer%Proliferation
目的 观察微小RNA(miRNA,miR)-10b对入胰腺癌细胞AsPC-1细胞(AsPC-1)增殖凋亡的影响.方法 重组真核表达载体pPG-CMV-EGFP-miR-10b(实验组)及空载体(对照组)转染AsPC-1细胞株.实时定量聚合酶链反应(Real-time PCR)和荧光显微镜检测载体瞬时表达的效率;细胞计数试剂盒(CCK-8)检测细胞生长曲线;流式细胞仪检测不同组间细胞的凋亡;Western blot检测细胞B细胞淋巴瘤/白血病-2蛋白家族促凋亡调节蛋白(Bim)、髓样细胞白血病-1(MCL-1)蛋白的表达.结果 与对照组(0.002 529±0.000 339)比较,实验组miR-10b表达量(0.020 500±0.007700)增加10倍,差异有统计学意义(P＜0.05).细胞生长曲线提示,实验组细胞增殖活性较对照组增高24％,差异有统计学意义(P＜0.05);实验组细胞凋亡率为3.0％,对照组为10.1％,差异有统计学意义(P＜0.05);实验组细胞Bim蛋白的相对表达水平(0.256 ±0.012)低于对照组(0.652±0.042),差异有统计学意义(P＜0.05);实验组细胞MCL-t蛋白的相对表达水平(0.515±0.025)高于对照组(0.154 ±0.068),差异均有统计学意义(P＜0.05).结论 miR-10b过表达可调控胰腺癌细胞Bim、MCL-1蛋白的表达,促进细胞增殖,抑制凋亡.
目的 觀察微小RNA(miRNA,miR)-10b對入胰腺癌細胞AsPC-1細胞(AsPC-1)增殖凋亡的影響.方法 重組真覈錶達載體pPG-CMV-EGFP-miR-10b(實驗組)及空載體(對照組)轉染AsPC-1細胞株.實時定量聚閤酶鏈反應(Real-time PCR)和熒光顯微鏡檢測載體瞬時錶達的效率;細胞計數試劑盒(CCK-8)檢測細胞生長麯線;流式細胞儀檢測不同組間細胞的凋亡;Western blot檢測細胞B細胞淋巴瘤/白血病-2蛋白傢族促凋亡調節蛋白(Bim)、髓樣細胞白血病-1(MCL-1)蛋白的錶達.結果 與對照組(0.002 529±0.000 339)比較,實驗組miR-10b錶達量(0.020 500±0.007700)增加10倍,差異有統計學意義(P＜0.05).細胞生長麯線提示,實驗組細胞增殖活性較對照組增高24％,差異有統計學意義(P＜0.05);實驗組細胞凋亡率為3.0％,對照組為10.1％,差異有統計學意義(P＜0.05);實驗組細胞Bim蛋白的相對錶達水平(0.256 ±0.012)低于對照組(0.652±0.042),差異有統計學意義(P＜0.05);實驗組細胞MCL-t蛋白的相對錶達水平(0.515±0.025)高于對照組(0.154 ±0.068),差異均有統計學意義(P＜0.05).結論 miR-10b過錶達可調控胰腺癌細胞Bim、MCL-1蛋白的錶達,促進細胞增殖,抑製凋亡.
목적 관찰미소RNA(miRNA,miR)-10b대입이선암세포AsPC-1세포(AsPC-1)증식조망적영향.방법 중조진핵표체재체pPG-CMV-EGFP-miR-10b(실험조)급공재체(대조조)전염AsPC-1세포주.실시정량취합매련반응(Real-time PCR)화형광현미경검측재체순시표체적효솔;세포계수시제합(CCK-8)검측세포생장곡선;류식세포의검측불동조간세포적조망;Western blot검측세포B세포림파류/백혈병-2단백가족촉조망조절단백(Bim)、수양세포백혈병-1(MCL-1)단백적표체.결과 여대조조(0.002 529±0.000 339)비교,실험조miR-10b표체량(0.020 500±0.007700)증가10배,차이유통계학의의(P＜0.05).세포생장곡선제시,실험조세포증식활성교대조조증고24％,차이유통계학의의(P＜0.05);실험조세포조망솔위3.0％,대조조위10.1％,차이유통계학의의(P＜0.05);실험조세포Bim단백적상대표체수평(0.256 ±0.012)저우대조조(0.652±0.042),차이유통계학의의(P＜0.05);실험조세포MCL-t단백적상대표체수평(0.515±0.025)고우대조조(0.154 ±0.068),차이균유통계학의의(P＜0.05).결론 miR-10b과표체가조공이선암세포Bim、MCL-1단백적표체,촉진세포증식,억제조망.
Objective To observe the effect of microRNA (miRNA,miR)-10b on the proliferation and apoptosis of human pancreatic cancer cell line AsPC-1.Methods The recombinant eukaryotic expression plasmid vector with miR-10b sequence (pPG-CMV-EGFP-miR-10b) was transfected into AsPC-1 cells with lipofectamine.The efficiency of the instant expression was confirmed by real-time quantitative polymerase chain reaction (Real-time PCR) and fluorescence microscope.The difference of the proliferation and apoptosis of the cells was evaluated by cell counting kit-8 (CCK-8) assay and flow cytometry.The expression of B-cell lymphoma/Leukemia-2 interacting mediator of cell death (Bim) and myeloid cell leukemia-1 (MCL-1) was detected by Western blotting.Results The expression of miR-10b in the sense group (0.020 500 ±0.007 700) was increased by 10 folds,as compared with the control (0.002 529 ±0.000 339,P ＜0.05).The ratio of cell proliferation in the experimenal group was 24％ higher than that of control group (P ＜ 0.05),and the apoptosis ratio in the experimenal group was 3.0％,lower than that of the control (10.1％).The expression of Bim and MCL-1 protein in the experimental group (0.256 ±0.012 and 0.515 ±0.025) was different with that of control (0.652 ±0.042 and 0.154 ± 0.068,P ＜ 0.05).Conclusion The overexpression of miR-10b can regulate the expression of Bim and MCL-1 protein,promote proliferation and inhibit the apoptosis of the AsPC-1 cells in vitro.
