Objective To investigated the effects of IL-32α on epithelial mesenchymal transition (EMT),metastasis and invasion of human pancreatic cancer cell lines Panc-1 and AsPC-1 and the underlining mechanism.Methods Human pancreatic cancer Panc-1 and AsPC-1 cells were treated with different concentrations of IL-32 α in vitro.EMT related marks E-cadherin,Vimentinand Zeb1,and extracellular matrix metalloproteinases (MMP-2 and MMP-9) were detected by immunofluorescence staining,Western blotting and real-time PCR,respectively.The activation of Jak2/STAT3 signaling proteins was detected by Western blotting.Results The RT-PCR and West blot showed that expressions of vimentin,Zeb1,MMP-2,MMP-9,p-Jak2,Jak2 and p-STAT3 mRNA and/or proteins in Panc-1 and AsPC-1 cells were inhibited after treated with IL-32α for 24h in a dose-dependent manner (P＜0.05);the expression of E-cadherin was upregulated (P＜0.05),while total STAT3 levels were not changed(P ＞0.05).The immunofluorescence assay showed that the expression of vimentin was increased after IL-32α treatment in a dose-dependent manner.Conclusion IL-32α can inhibit the epithelial mesenchymal transition,reduce secretion of MMPs in a dose-dependent manner inPanc-1 and AsPC-1 cells,which is associated with the deactivation of Jak2/STAT3 signaling pathway.%目的 观察IL-32 α作用于人胰腺癌Panc-1、AsPC-1细胞后上皮间质转化(EMT)、侵袭转移及Jak2/STAT3信号通路的改变,并探讨其可能的作用机制.方法 采用RT-PCR、Western blot及细胞免疫荧光技术,检测不同浓度IL-32α处理24h后胰腺癌Panc-1、AsPC-1细胞的EMT相关标志物(E-cadherin、Vimentin、Zeb1)、侵袭转移相关分子标志物(MMP-2、MMP-9)mRNA与蛋白以及Jak2/STAT3信号通路相关蛋白p-Jak2、Jak2、p-STAT3、STAT3的表达情况.结果 RT-PCR及Western blot均显示胰腺癌Panc-1、AsPC-1细胞中E-cadherin的mRNA与蛋白表达水平均呈IL-32α剂量依赖性上调(均P＜0.05);Vimentin、Zeb1、MMP-2及MMP-9的mRNA与蛋白表达水平均呈IL-32α剂量依赖性下调(均P＜0.05);p-Jak2、Jak2、p-STAT3的表达水平均呈IL-32 α剂量依赖性下调(均P＜0.05),STAT3无明显改变(P＞0.05).细胞免疫荧光染色显示Vimentin蛋白表达于细胞质中,且荧光强度呈IL-32 α剂量依赖性下调(均P＜0.05).结论 IL-32α在一定剂量范围内以剂量依赖性的方式抑制胰腺癌Panc-1、AsPC-1细胞的EMT和侵袭转移,可能与抑制Jak2/STAT3信号通路有关.
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