CAJ | 학술논문

目的:研究用RNA干扰(RNA interference,RNAi)技术抑制转录因子Snail表达后,对人肺腺癌A549细胞上皮-间充质转分化表型和体外侵袭能力的影响.方法:构建能表达针对Snail的小干扰RNA(Small interfering RNA,siRNA)的RNA干扰载体(Snail siRNA vector)和表达不针对任何已知mRNA的siRNA的阴性对照RNA干扰载体(control siRNA vector),分别转染A549细胞,新霉素抗性筛选得到Snail表达受抑制的A549-siSnail细胞和Snail表达未受影响的A549-siControl细胞.针对非转染(A549-nontransfection)、A549-siSnail、A549-siControl三组细胞,分别采用RT-PCR和Western blot技术检测Snail、α-平滑肌肌动蛋白(α-SMA)和E-钙粘素表达,用Boyden chamber模型检测细胞侵袭能力.结果: A549-nontransfection组:Snail和α-SMA表达强阳性,E-钙粘素表达弱阳性；A549-siSnail组:和A549-nontransfection组相比,Snail和α-SMA表达显著减弱(P<0.01),E-钙粘素表达显著增强(P<0.01),Boyden chamber穿膜细胞数显著减少(P<0.01)；A549-siControl组:Snail、α-SMA和E-钙粘素表达,及Boyden chamber穿膜细胞数和A549-nontransfection组无显著差异(P>0.05).结论: 通过RNA干扰阻滞Snail表达能有效地抑制A549细胞上皮-间充质转分化及体外侵袭能力.Snail可能在肺腺癌上皮-间充质转分化及侵袭过程中扮演重要角色,抑制Snail表达可能成肺腺癌治疗的可行策略.
목적:연구용RNA간우(RNA interference,RNAi)기술억제전록인자Snail표체후,대인폐선암A549세포상피-간충질전분화표형화체외침습능력적영향.방법:구건능표체침대Snail적소간우RNA(Small interfering RNA,siRNA)적RNA간우재체(Snail siRNA vector)화표체불침대임하이지mRNA적siRNA적음성대조RNA간우재체(control siRNA vector),분별전염A549세포,신매소항성사선득도Snail표체수억제적A549-siSnail세포화Snail표체미수영향적A549-siControl세포.침대비전염(A549-nontransfection)、A549-siSnail、A549-siControl삼조세포,분별채용RT-PCR화Western blot기술검측Snail、α-평활기기동단백(α-SMA)화E-개점소표체,용Boyden chamber모형검측세포침습능력.결과: A549-nontransfection조:Snail화α-SMA표체강양성,E-개점소표체약양성；A549-siSnail조:화A549-nontransfection조상비,Snail화α-SMA표체현저감약(P<0.01),E-개점소표체현저증강(P<0.01),Boyden chamber천막세포수현저감소(P<0.01)；A549-siControl조:Snail、α-SMA화E-개점소표체,급Boyden chamber천막세포수화A549-nontransfection조무현저차이(P>0.05).결론: 통과RNA간우조체Snail표체능유효지억제A549세포상피-간충질전분화급체외침습능력.Snail가능재폐선암상피-간충질전분화급침습과정중분연중요각색,억제Snail표체가능성폐선암치료적가행책략.