RNA干扰抑制Snail表达对于胃癌细胞上皮间充质转化以及体外侵袭的影响

INFLUENCE OF INHIBITION OF Snail EXPRESSION BY RNA INTERFERENCE ON EPITHELIAL-MESENCHYMAL TRANSITION AND INVASION ABILITY OF HUMAN GASTRIC CARCINOMA CELL SGC-7901 IN VITRO

目的用RNA干扰(RNAinterference,RNAi)技术抑制转录因子Snail表达,观察其对人胃腺癌SGC-7901细胞上皮-间充质转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 interferingRNA,si RNA)的RNA干扰载体(Snail si RNAvector)和表达不针对任何已知mRNA的si RNA的阴性对照RNA干扰载体(control si RNAvector),分别转染SGC-7901细胞,筛选得到Snail表达受抑制的SGC-7901-siSnail细胞和Snail表达未受影响的SGC-7901-siControl细胞。分别采用RT-PCR和Western blot技术检测非转染组、SGC-7901-siSnail、SGC-7901-siControl三组细胞Snail、α-平滑肌肌动蛋白(α-SMA)和E-cadherin表达,用Boyden chamber模型检测细胞侵袭能力。结果 SGC-7901-siSnail组与SGC-7901-nontransfection组相比,Snail和α-SMA表达显著减弱(P<0.01),E-cadherin表达显著增强(P<0.01),Boyden chamber穿膜细胞数显著减少(P<0.01);SGC-7901-siControl组中Snail、α-SMA、E-cadherin表达、Boyden chamber穿膜细胞数分别和SGC-7901-nontransfection组比较无显著差异(P>0.05)。结论通过RNA干扰阻滞Snail表达能有效地抑制SGC-7901细胞上皮-间充质转化及体外侵袭能力。Snail可能在胃腺癌上皮-间充质转化及侵袭过程中扮演重要角色,抑制Snail表达可能成胃腺癌治疗的可行策略。

Objective To investigate the effect of RNA interference（RNAi） targeting transcription factor Snail on epithelial-mesenchymal transition（EMT） and invasion ability of human gastric carcinoma cell SGC-7901 in vitro.Methods RNA interference plasmids that express small interfering RNA（siRNA） targeting Snail（Snail siRNA vector） or express siRNA that do not match any known human coding mRNA（control siRNA vector）were designed,constructed,and lipotransfected into SGC-7901 cell line.SGC-7901-si Snail cells expressing suppressed Snail or SGC-7901-si-Control cells expressing uninfluenced Snail was selected by neomycin resistance.In SGC-7901-nontransfection,SGC-7901-siSnail and SGC-7901-siControl groups,Snail,alpha-smooth muscle actin（α-SMA） and E-cadherin expression were examined by reverse transcription-polymerase chain reaction（RT-PCR） and Western blot,and the invasion ability was examined by the Boyden chamber model.Results SGC-7901-nontransfection group： Both Snail and α–SMA expressions were strongly positive,but E-cadherin expression was weakly.SGC-7901-siSnail group： Compared with those in the SGC-7901-nontransfection group,Snail and α-SMA expressions,and the numbers of SGC-7901 cells permeating the septum of Boyden chamber（NCS） were decreased,but E-cadherin expression was increased significantly（P〈0.01）.SGC-7901-siControl group： Compared with those in the SGC-7901-nontransfection group,Snail,α-SMA and E-cadherin expression,and NCS were similar.Conclusion RNAi targeting Snail can inhibit EMT and invasion of SGC-7901 cells efficiently in vitro.Snail might play a crucial role in EMT and invasion of lung carcinoma,and suppression of Snail expression might be a promising strategy for the treatment of human lung carcinoma.