国际肿瘤学杂志
國際腫瘤學雜誌
국제종류학잡지
JOURNAL OF INTERNATIONAL ONCOLOGY
2013年
6期
476-479
,共4页
杜叶平%武春梅%房树志%吴洁%苗晋华
杜葉平%武春梅%房樹誌%吳潔%苗晉華
두협평%무춘매%방수지%오길%묘진화
微RNAs%结肠肿瘤%细胞增殖%细胞迁移分析
微RNAs%結腸腫瘤%細胞增殖%細胞遷移分析
미RNAs%결장종류%세포증식%세포천이분석
MicroRNAs%Colonic neoplasms%Cell proliferation%Cell migration assays
目的 研究miR-200b对人结肠癌sw620细胞E-cadherin表达的调节作用,探讨miR-200b对sw620细胞增殖及迁移的影响.方法 采用实时PCR (RT-PCR)技术分别检测转染pEGP-miR-200b后24h和72 h sw620细胞中miR-200b的表达变化;采用RT-PCR技术和Western blot方法检测转染miR-200b后对E-cadherin表达的影响;采用四甲基偶氮唑蓝法(MTT)和划痕擦伤实验检测转染miR-200b对sw620细胞增殖、迁移的影响.结果 miR-200b转染sw620细胞48 h和72 h后,miR-200b相对表达量较对照组显著上调(t=11.579,P＜0.01;t=11.579,P＜0.01);且抑制了细胞的增殖能力,第3天抑制作用最显著,抑制率为55.34％;细胞的迁移速度受到明显的抑制,两组在24、48、72 h的迁移距离有显著差异(t=11.579,P＜0.01;t=10.419,P＜0.01;t=6.955,P＜0.01).同时细胞中E-cadherin基因的mRNA和蛋白表达水平显著升高(t=10.432,P＜0.01;t=8.325,P＜0.01).结论 上调miR-200b表达可抑制结肠癌细胞的生物活性,该作用可能与E-cadherin的表达有关.
目的 研究miR-200b對人結腸癌sw620細胞E-cadherin錶達的調節作用,探討miR-200b對sw620細胞增殖及遷移的影響.方法 採用實時PCR (RT-PCR)技術分彆檢測轉染pEGP-miR-200b後24h和72 h sw620細胞中miR-200b的錶達變化;採用RT-PCR技術和Western blot方法檢測轉染miR-200b後對E-cadherin錶達的影響;採用四甲基偶氮唑藍法(MTT)和劃痕抆傷實驗檢測轉染miR-200b對sw620細胞增殖、遷移的影響.結果 miR-200b轉染sw620細胞48 h和72 h後,miR-200b相對錶達量較對照組顯著上調(t=11.579,P＜0.01;t=11.579,P＜0.01);且抑製瞭細胞的增殖能力,第3天抑製作用最顯著,抑製率為55.34％;細胞的遷移速度受到明顯的抑製,兩組在24、48、72 h的遷移距離有顯著差異(t=11.579,P＜0.01;t=10.419,P＜0.01;t=6.955,P＜0.01).同時細胞中E-cadherin基因的mRNA和蛋白錶達水平顯著升高(t=10.432,P＜0.01;t=8.325,P＜0.01).結論 上調miR-200b錶達可抑製結腸癌細胞的生物活性,該作用可能與E-cadherin的錶達有關.
목적 연구miR-200b대인결장암sw620세포E-cadherin표체적조절작용,탐토miR-200b대sw620세포증식급천이적영향.방법 채용실시PCR (RT-PCR)기술분별검측전염pEGP-miR-200b후24h화72 h sw620세포중miR-200b적표체변화;채용RT-PCR기술화Western blot방법검측전염miR-200b후대E-cadherin표체적영향;채용사갑기우담서람법(MTT)화화흔찰상실험검측전염miR-200b대sw620세포증식、천이적영향.결과 miR-200b전염sw620세포48 h화72 h후,miR-200b상대표체량교대조조현저상조(t=11.579,P＜0.01;t=11.579,P＜0.01);차억제료세포적증식능력,제3천억제작용최현저,억제솔위55.34％;세포적천이속도수도명현적억제,량조재24、48、72 h적천이거리유현저차이(t=11.579,P＜0.01;t=10.419,P＜0.01;t=6.955,P＜0.01).동시세포중E-cadherin기인적mRNA화단백표체수평현저승고(t=10.432,P＜0.01;t=8.325,P＜0.01).결론 상조miR-200b표체가억제결장암세포적생물활성,해작용가능여E-cadherin적표체유관.
Objective To study the effects of miR-200b on proliferation and migration of sw620 colon cancer cells,and its regulation effect on E-cadherin expression.Methods The expressions of miR-200b in sw620 cells at 24 h and 72 h after pEGP-miR-200b transfection were detected by real-time PCR (RT-PCR).The change of the expression level of E-cadherin after miR-200b transfection was detected using the methods of RT-PCR and Western blot.The proliferation and migration abilities were measured by MTT and scratch test after miR-200b transfection.Results The expressions of miR-200b in sw620 cells at 24 h and 72 h after pEGP-miR-200b transfection raised significantly compared to the control group (t =11.579,P ＜ 0.01 ; t =11.579,P ＜0.01).MiR-200b transfection inhibited the proliferation abilities of sw620 cells.It is the most significant of the inhibitory effect on the third day and the inhibition rate was 55.34％.MiR-200b transfection markedly inhibited the migration abilities of sw620 cells.The two groups had significant difference in the migration distance of 24,48,72 h (t =11.579,P ＜0.01 ; t =10.419,P ＜0.01 ; t =6.955,P ＜0.01).The mRNA and protein expressions of E-cadherin gene increased significantly by transfecting miR-200b gene in sw620 cells (t =10.432,P ＜ 0.01 ; t =8.325,P ＜ 0.O1).Conclusion Up-regulated expression of miR-200b could inhibite the proliferation and migration abilities of sw620 colon cancer cells.The involved molecular mechanism is probably related to the change of E-cadherin expression.
