中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2015年
8期
1899-1901
,共3页
王公平%周博%金灿辉%汪增方%杨言通%陈晔%张伟%王珍珍%裴哲
王公平%週博%金燦輝%汪增方%楊言通%陳曄%張偉%王珍珍%裴哲
왕공평%주박%금찬휘%왕증방%양언통%진엽%장위%왕진진%배철
微小RNA-9%胃癌%SGC-7901%增殖%凋亡
微小RNA-9%胃癌%SGC-7901%增殖%凋亡
미소RNA-9%위암%SGC-7901%증식%조망
MicroRNA-9%Gastri caner%SGC-7901%Proliferation%Apoptosis.
目的 观察微小RNA(miRNA,miR)-9对胃癌细胞增殖及凋亡的影响,以及对B细胞淋巴瘤/白血病-2(bcl-2)及bcl-2相关X蛋白(bax)蛋白表达的调控作用.方法 将50 nmol/L的miR-9 mimics及阴性对照序列由Lipofectamine 2000转染入人胃癌SGC-7901细胞中;实时定量反转录聚合酶链反应(RT-qPCR)法检测转染后胃癌细胞内miR-9表达水平;细胞计数试剂盒(CCK-8)法检测细胞增殖能力;划痕实验检测细胞迁移能力;流式细胞术分析转染后细胞凋亡的变化;Western blot法检测bcl-2、bax蛋白表达水平.结果 转染24h,miR-9 mimics转染组细胞内miR-9表达水平较空白对照组明显升高,约为空白对照组的(45.69±2.67)倍,差异有统计学意义(F=179.45,P＜0.05);与空白对照组比较,miR-9 mimics转染组细胞增殖受抑制(P＜0.05),24、48、72 h细胞增殖抑制率分别为18.45％、35.12％、53.78％,凋亡细胞增多(P＜0.05),bcl-2蛋白表达下调,bax表达上调.结论 miR-9能够抑制SGC7901细胞增殖并诱导胃癌细胞凋亡,这可能与其抑制bcl-2蛋白的表达有关.
目的 觀察微小RNA(miRNA,miR)-9對胃癌細胞增殖及凋亡的影響,以及對B細胞淋巴瘤/白血病-2(bcl-2)及bcl-2相關X蛋白(bax)蛋白錶達的調控作用.方法 將50 nmol/L的miR-9 mimics及陰性對照序列由Lipofectamine 2000轉染入人胃癌SGC-7901細胞中;實時定量反轉錄聚閤酶鏈反應(RT-qPCR)法檢測轉染後胃癌細胞內miR-9錶達水平;細胞計數試劑盒(CCK-8)法檢測細胞增殖能力;劃痕實驗檢測細胞遷移能力;流式細胞術分析轉染後細胞凋亡的變化;Western blot法檢測bcl-2、bax蛋白錶達水平.結果 轉染24h,miR-9 mimics轉染組細胞內miR-9錶達水平較空白對照組明顯升高,約為空白對照組的(45.69±2.67)倍,差異有統計學意義(F=179.45,P＜0.05);與空白對照組比較,miR-9 mimics轉染組細胞增殖受抑製(P＜0.05),24、48、72 h細胞增殖抑製率分彆為18.45％、35.12％、53.78％,凋亡細胞增多(P＜0.05),bcl-2蛋白錶達下調,bax錶達上調.結論 miR-9能夠抑製SGC7901細胞增殖併誘導胃癌細胞凋亡,這可能與其抑製bcl-2蛋白的錶達有關.
목적 관찰미소RNA(miRNA,miR)-9대위암세포증식급조망적영향,이급대B세포림파류/백혈병-2(bcl-2)급bcl-2상관X단백(bax)단백표체적조공작용.방법 장50 nmol/L적miR-9 mimics급음성대조서렬유Lipofectamine 2000전염입인위암SGC-7901세포중;실시정량반전록취합매련반응(RT-qPCR)법검측전염후위암세포내miR-9표체수평;세포계수시제합(CCK-8)법검측세포증식능력;화흔실험검측세포천이능력;류식세포술분석전염후세포조망적변화;Western blot법검측bcl-2、bax단백표체수평.결과 전염24h,miR-9 mimics전염조세포내miR-9표체수평교공백대조조명현승고,약위공백대조조적(45.69±2.67)배,차이유통계학의의(F=179.45,P＜0.05);여공백대조조비교,miR-9 mimics전염조세포증식수억제(P＜0.05),24、48、72 h세포증식억제솔분별위18.45％、35.12％、53.78％,조망세포증다(P＜0.05),bcl-2단백표체하조,bax표체상조.결론 miR-9능구억제SGC7901세포증식병유도위암세포조망,저가능여기억제bcl-2단백적표체유관.
Objective To investigate the effect of microRNA (miR)-9 on proliferation and apoptosis of gastric cancer cells,and on the regulation of expression of bcl-2 and bax protein.Methods MiR-9 mimics and 50 nmol/L negative control sequence is composed of 2000/L Lipofectamine was transfected into human gastric cancer SGC-7901 cells;real-time reverse transcriptase-polymerase chain reaction (RT-qPCR) was detected after transfection in gastric cancer cells miR-9 expression level;ability of cell counting kit-8 (CCK-8) method to detect cell proliferation;changes of apoptosis after transfection were analyzed by flow cytometry;detection of bcl-2 Western blotting method,the expression of bax protein the level of.Results 24 cells were transfected with H,miR-9 mimics expression level of miR-26 in cells transfected group compared with blank control group increased significantly (P ＜ 0.05);Compared with the blank control group,cell proliferation was inhibited in miR-26mimics transfection group (P ＜ 0.05),the apoptotic cells increased (P ＜ 0.05),bcl-2 protein expression down regulated,the up regulation of bax expression.Conclusion MiR-26 can inhibit the proliferation of SGC7901 cells and induce apoptosis of gastric cancer cells,the expression of which may be related to the inhibition of bcl-2 protein.
