【摘要】 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。更多还原

【Abstract】 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-32α 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 24 h 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 ofMMPs in a dose-dependent manner inPanc-1 and AsPC-1 cells, which is associated with the deactivation of Jak2/STAT3 signaling pathway.更多还原