CAJ | 학술논문

目的探讨沉默信号传导与转录激活基因3(STAT3)对人胰腺癌细胞系SW1990上皮间质转换的影响及机制.方法应用RNA干扰技术,建立STAT3基因稳定沉默的胰腺癌细胞系SW1990.MTT法检测细胞的增殖情况,流式细胞仪检测细胞周期.体外细胞侵袭实验观察侵袭能力的改变.实时PCR、蛋白印迹方法检测STAT3、P-STAT3、TWIST、Snail和E-cadherin基因的mRNA、蛋白含量.结果 STAT3基因沉默后,细胞增殖水平下降(P＜0.05),细胞侵袭能力下降.实时PCR、蛋白印迹结果显示STAT3、P-STAT3蛋白表达下降,TWIST基因表达量极少,Snail mRNA和蛋白表达明显下调(P＜0.05),E-cadherin明显上调(P＜0.05).结论 STAT3信号通路在胰腺癌上皮间质转换中起着重要作用.沉默STAT3基因可抑制人胰腺癌细胞上皮间质转换过程.
목적 탐토침묵신호전도여전록격활기인3(STAT3)대인이선암세포계SW1990상피간질전환적영향급궤제.방법 응용RNA간우기술,건립STAT3기인은정침묵적이선암세포계SW1990.MTT법검측세포적증식정황,류식세포의검측세포주기.체외세포침습실험관찰침습능력적개변.실시PCR、단백인적방법검측STAT3、P-STAT3、TWIST、Snail화E-cadherin기인적mRNA、단백함량.결과 STAT3기인침묵후,세포증식수평하강(P＜0.05),세포침습능력하강.실시PCR、단백인적결과현시STAT3、P-STAT3단백표체하강,TWIST기인표체량겁소,Snail mRNA화단백표체명현하조(P＜0.05),E-cadherin명현상조(P＜0.05).결론 STAT3신호통로재이선암상피간질전환중기착중요작용.침묵STAT3기인가억제인이선암세포상피간질전환과정.
Objective To investigate the effect and mechanism of RNAi-mediated STAT3 gene silencing on epithelial-to-mesenchymal transition (EMT) of human pancreatic cancer cells.Methods Lentivirus vector mediating RNA interference targeting STAT3 was constructed in SW1990 cell line.The invasion ability of SW1990 cells was determined by cell invasion assay in vitro.Cell proliferation and cell cycle of SW1990 cells were also detected.The expression of EMT related genes such as STAT3,P-STAT3,Twist,Snail and E-cadherin were analyzed by reverse transcription-PCR,real-time PCR,and Western blotting.Results Silencing of STAT3 with RNAi not only markedly reduced proliferation but also greatly decreased the invasion ability of SW1990 cells.The mRNA level and protein expression of Snail decreased significantly (P ＜ 0.05),but those of E-cadherin increased significantly (P ＜ 0.05),compared to parental cells.However,no difference was on the expression of Twist in SW1990 cell line.Conclusions STAT3 signaling pathway plays an important role in the process of EMT.Silencing of STAT3 with RNAi can significantly inhibit EMT by downregulating expression of Snail and E-cadherin in pancreatic cancer cells.