山西医科大学学报2025，Vol.56Issue(4)：369-379,11.DOI:10.13753/j.issn.1007-6611.2025.04.005
胰腺星状细胞Cav-1敲低调控Shh通路促进胰腺癌细胞增殖
Cav-1 knockdown in pancreatic stellate cells promotes pancreatic cancer cell proliferation through Shh signaling
摘要
Abstract
Objective To investigate the effect and mechanism of Caveolin-1(Cav-1)expression in pancreatic stellate cells(PSCs)on the proliferation of pancreatic cancer cells.Methods In vivo,Aspc-1 cells with Gli-1 knockdown and PSCs with Cav-1 knock-down were implanted subcutaneously into nude mice.The growth of tumor cells in vivo was measured.The vascular density MVD in tu-mor tissues was detected by immunohistochemical staining of CD31 and the protein expression of CD31 in tumor tissues was detected by Western blot.In vitro,Cav-1 expression in PSCs was knocked down to investigate the changes of PSCs cytokine secretion.After Gli-1-knockdown Aspc-1 cells were co-cultured with PSCs,and then treated with cyclopamine(a Hedgehog pathway SMO factor inhibitor)or n-acetylcysteine(NAC),and then the invasion and proliferation capacities of Aspc-1 cells were investigated.Nrf2 expression in PSCs was overexpressed to investigate the changes of cytokine secretion and ROS production in PSCs.ELISA was used to determine the secretion of Shh,MMP2,bFGF and IL-6 in PSCs.Western blot was performed to detect the protein expression levels of Shh,Gli-1,CyclinA1,CyclinD1 and Nrf2 in PSCs and Aspc-1 cells.The invasion ability of Aspc-1 was detected by Transwell,and the prolif-eration ability of Aspc-1 cells was detected by MTT and[3H]thymine incorporation assay.The tubule formation ability of human um-bilical vein endothelial cells(HUVECs)was detected.The secretions of Shh,MMP2,bFGF and IL-6 in PSCs were determined by ELISA.Results In vivo,the tumor volume,MVD and CD31 protein expression were significantly increased in nude mice after incu-bated with Cav-1-knockdown PSCs(P<0.05).When the expression of Gli-1 was knocked down in Aspc-1,the elevated tumor growth rate induced by Cav-1-knockdown PSCs was significantly decreased(P<0.05).In vitro,after Cav-1 was knocked down in PSCs,the se-cretions of Shh,MMP2,bFGF and IL-6 in PSCs supernatant were significantly increased(P<0.05),the protein expression of Nrf2 was significantly increased(P<0.05),the expression levels of Gli-1 protein,CyclinA1 and CyclinD1 proteins in Aspc-1 cells were signifi-cantly increased(P<0.05),and the invasion and proliferation abilities of Aspc-1 cells were also significantly increased(P<0.05).Cy-clopamine significantly inhibited the elevated proliferation rate of Aspc-1 cells induced by Cav-1-knockdown PSCs(P<0.05).NAC sig-nificantly decreased the upregulated expression of Gli-1 in Aspc-1 induced by Cav-1-knockdown PSCs(P<0.05),and inhibited the tu-bule formation ability of HUVECs induced by Cav-1-knockdown PSCs(P<0.05).Overexpression of Nrf2 in PSCs inhibited the secre-tions of Shh,MMP2,bFGF and IL-6 in Cav-1-knockdown PSCs supernatant(P<0.05),and decreased the expression of Shh protein in Cav-1-knockdown PSCs(P<0.05).Conclusion Cav-1 knockdown in PSCs could activate Shh pathway by downregulating the expres-sion of Nrf2 to promote the proliferation of pancreatic cancer cells.关键词
胰腺星状细胞/Caveolin-1/核因子红细胞相关因子2(Nrf2)/Shh信号通路/胰腺癌/细胞增殖Key words
pancreatic stellate cells/Caveolin-1/Nrf2/Shh signaling/pancreatic cancer/cell proliferation分类
临床医学引用本文复制引用
邵珊,赵新汉,雷建军..胰腺星状细胞Cav-1敲低调控Shh通路促进胰腺癌细胞增殖[J].山西医科大学学报,2025,56(4):369-379,11.基金项目
国家自然科学基金资助项目(81502066,81702908) （81502066,81702908）
陕西省自然科学基础研究计划(面上)项目(2019JM-115,2018JQ8030) （面上）
西安交通大学基础科研业务费(xzy012019090) （xzy012019090）
