IL-32α抑制胰腺癌细胞上皮间质转化和侵袭转移作用机制研究

Effect of IL-32α on epithelial-mesenchymal transition and invasion/metastasis of pancreatic cancer cells and its mechanism

目的观察IL-32α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-32α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-32α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。

Objective To investigated the effects of IL-32αon epithelial mesenchymal transition(EMT),metastasis and invasion of human pancreatic cancer cell lines Panc-1and AsPC-1and the underlining mechanism.Methods Human pancreatic cancer Panc-1and AsPC-1cells were treated with different concentrations of IL-32αin vitro.EMT related marks E-cadherin,Vimentinand Zeb1,and extracellular matrix metalloproteinases(MMP-2and MMP-9)were detected by immunofluorescence staining,Western blotting and real-time PCR,respectively.The activation of Jak2/STAT3signaling 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,Jak2and p-STAT3mRNA and/or proteins in Panc-1and AsPC-1cells were inhibited after treated with IL-32αfor24h in a dose-dependent manner(P<0.05);the expression of E-cadherin was upregulated(P<0.05),while total STAT3levels were not changed(P>0.05).The immunofluorescence assay showed that the expression of vimentin was increased after IL-32αtreatment in a dose-dependent manner.Conclusion IL-32αcan inhibit the epithelial mesenchymal transition,reduce secretion of MMPs in a dose-dependent manner inPanc-1and AsPC-1cells,which is associated with the deactivation of Jak2/STAT3signaling pathway.