缺氧激活胰腺星状细胞促进胰腺癌进展

Hypoxia activates PSCs to promote the progression of pancreatic cancer

目的探讨缺氧在星状细胞活化及胰腺癌进展中的重要作用。方法通过缺氧培养胰腺星状细胞(PSCs)及共培养PSCs与胰腺癌Panc-1细胞,以常氧培养作为对照,免疫荧光检测各组PSCs的活化状态,ELISA检测各组PSCs白介素-6(IL-6)、基质衍生因子-1(SDF-1)、血管内皮生长因子A(VEGF-A)的分泌量,Westernblot检测胰腺癌细胞E-cadherin和Vimentin蛋白表达,Transwell小室侵袭实验检测各组Panc-1细胞的侵袭能力。HIF-1αshRNA稳定转染胰腺癌细胞,免疫荧光检测各组PSCs的活化状态,ELISA检测各组PSCsIL-6、SDF-1、VEGF-A的分泌量。结果缺氧可激活胰腺星状细胞,增加星状细胞IL-6、SDF-1、VEGF-A的分泌量,明显增加星状细胞,促进胰腺癌Panc-1细胞的侵袭能力,并上调HIF-1α和Vimentin蛋白,下调E-cadherin蛋白。靶向沉默HIF-1α基因后,缺氧失去了活化胰腺星状细胞及胰腺癌细胞侵袭和EMT的诱导作用。结论缺氧通过HIF-1α激活胰腺星状细胞诱导胰腺癌细胞侵袭及细胞上皮-间质转分化过程。

Objective To investigate the effects of hypoxia on the activation of pancreatic stellate cells (PSCs) and progression of pancreatic cancer. Methods In the present study,PSCs were cultured under normoxic or hypoxic conditions or co-cultured with pancreatic cancer cell line Panc-1. Then,the shRNA of HIF- 1α was stably transfected into PSC cells. Activation of PSCs was detected by immunofluorescence. Secretions of IL-6,SDF-1 and VEGF-A in PSCs were determined by ELISA assay. Invasion of Panc-1 was detected by the Transwell assay. The proteins of E-cadherin and Vimentin in Panc-1 cells were determined by Western blot. Results Hypoxia activated PSCs and significantly increased IL-6,SDF-1 and VEGF-A secretion in PSCs. Moreover,hypoxia significantly upregulated the PSCs-induced invasion of Panc-1 cells,increased the protein expressions of HIF- 1α and Vimentin,and decreased the protein expression of E-cadherin. Furthermore,knockdown of HIF- 1α obviously abrogated hypoxia-induced PSC activation and IL-6,SDF-1 and VEGF-A secretion in PSCs,and abolished hypoxia-enhanced epithelial-mesenchymal transition and invasion of Panc-1 cells. Conclusion Hypoxia enhances the epithelial-mesenchymal transition and invasion of PSCs and strengthens HIF- 1α expression via activating PSCs.