胰腺癌相关性成纤维细胞中前列腺环素合成酶对胰腺癌细胞作用研究  

Effects and mechanisms of PGIS in pancreatic cancer-associated fibroblasts on proliferation,migration,and invasion of pancreatic cancer cells

作　　者：潘越 刘勇 李壮 朱峰 PAN Yue;LIU Yong;LI Zhuang;ZHU Feng(Department of General Surgery,Sir RunRun Hospital,Nanjing Medical University,Nanjing 211112,China)

机构地区：[1]南京医科大学附属逸夫医院普外科,江苏南京211112

出　　处：《中国肿瘤外科杂志》2023年第5期476-485,共10页Chinese Journal of Surgical Oncology

基　　金：南京市卫健委医学科技发展基金项目(YKK19172)。

摘　　要：目的探究胰腺癌相关性成纤维细胞(CAFs)中的前列腺环素合成酶(PGIS/PTGIS)对胰腺癌细胞增殖、迁移和侵袭能力的影响及相关作用机制。方法利用GEPIA、Kaplan-Meier等数据库分析PGIS在胰腺癌及正常胰腺组织中的表达差异;通过q-PCR检测PGIS在CAFs及人胰腺星状细胞(HPSC)中的表达差异;通过CoCl_(2)化学诱导构建肿瘤微环境缺氧模型,检测胰腺癌及CAFs细胞的PGIS表达水平;利用siRNA转染、慢病毒感染的方式分别构建敲低和过表达PGIS的CAFs细胞,通过CCK-8增殖、Transwell迁移及侵袭等实验探究敲低和过表达PGIS的CAFs细胞对胰腺癌细胞增殖、迁移和侵袭能力的影响;通过小鼠胰腺原位成瘤探究过表达PGIS的CAFs细胞在体内对胰腺癌细胞增殖的影响;通过Western blot检测过表达PGIS的CAFs细胞对胰腺癌PI3K/Akt通路相关蛋白表达的影响。结果GEPIA、Kaplan-Meier数据库分析显示,与正常胰腺组织相比,PGIS在胰腺癌组织中高表达,并且与患者不良预后相关(P<0.05);与HPSC细胞相比,PGIS在CAFs细胞中显著高表达(P<0.05);胰腺癌及CAFs细胞在缺氧微环境中PGIS的表达水平明显上调(P<0.05);与对照组相比,敲低CAFs中的PGIS后,胰腺癌细胞的增殖速率明显减慢,细胞迁移和侵袭数量均明显减少(P<0.05);与对照组相比,过表达CAFs中的PGIS后,胰腺癌细胞的增殖速率明显加快,细胞迁移和侵袭数量均明显增多(P<0.05);与对照组相比,过表达PGIS的CAFs在体内可促进胰腺肿瘤的生长(P<0.05);与对照组相比,过表达CAFs中的PGIS后可明显上调VEGFA、p-Akt的蛋白表达水平(P<0.05),也可明显上调FGF-2、α-SMA及MMP-9的mRNA水平(P<0.05)。结论PGIS在胰腺癌组织中呈高表达,CAFs中的PGIS可能通过调节PI3K/Akt信号通路正向调控VEGFA的表达上调,促进胰腺癌细胞的增殖和侵袭,参与胰腺癌的发生、发展。Objective To explore the effects and related mechanisms of prostaglandin synthetase(PGIS/PTGIS)in pancreatic cancer-associated fibroblasts(CAFs)on the proliferation,migration,and invasion abilities of pancreatic cancer cells.Methods The expression difference of PGIS in pancreatic cancer and normal tissues was analyzed using GEPIA and Kaplan-Meier databases.The expression of PGIS in CAFs and human pancreatic stellate cells(HPSC)was detected by q-PCR.The hypoxia model of tumor microenvironment was constructed by chemical induction of CoCl_(2),and the PGIS expression level of pancreatic cancer and CAFs cells was detected.Lentivirus infection and siRNA transfection were used to construct CAFs cells with knockdown and overexpression of PGIS.The effects of CAFs cells with knockdown and overexpression of PGIS on the proliferation,migration,and invasion of pancreatic cancer cells were investigated by CCK-8 proliferation,Transwell migration,and invasion experiments.The effect of CAFs cells overexpressing PGIS on the proliferation of pancreatic cancer cells in vivo was investigated by in situ tumor formation of mouse pancreas.The effect of CAFs cells overexpressing PGIS on the expression of PI3K/Akt pathway related proteins in pancreatic cancer was detected by Western blot.Results GEPIA and Kaplan-Meier database analysis showed that compared with normal pancreatic tissues,PGIS was highly expressed in pancreatic cancer tissues and correlated with poor prognosis(P<0.05).Compared with HPSC cells,PGIS was significantly overexpressed in CAFs cells(P<0.05).The expression level of PGIS in pancreatic cancer and CAFs cells was significantly up-regulated in anoxic microenvironment(P<0.05).Compared with the control group,knocking down PGIS in CAFs significantly slowed down the proliferation rate of pancreatic cancer cells,and the number of cell migration and invasion were reduced significantly(P<0.05).Compared with the control group,overexpression of PGIS in CAFs significantly accelerated the proliferation rate of pancreatic cancer c

关 键 词：胰腺癌 前列腺环素合成酶 癌相关性成纤维细胞 PI3K/Akt信号通路 VEGFA 

分 类 号：R735.9[医药卫生—肿瘤]