肿瘤相关成纤维细胞来源外泌体ciRS-7经miR-7/IGF1R轴促进胰腺癌细胞糖酵解

The inhibiting effect of exosomal ciRS-7 derived from cancer associated fibroblasts on glycolysis of pancreatic cancer cells via miR-7/IGF1R axis

目的:探讨肿瘤相关成纤维细胞(CAFs)来源外泌体ciRS-7对胰腺癌细胞糖酵解代谢的影响。方法:采用超速离心法提取CAFs外泌体,并通过实时定量聚合酶链反应(RT-qPCR)检测外泌体中ciRS-7的表达及胰腺癌细胞糖酵解关键酶mRNA的表达;构建含ciRS-7野生及突变序列的荧光素酶检测报告质粒,在胰腺癌细胞中验证ciRS-7与miR-7的关系;蛋白质印迹(Western blot)法检测IGF1R/AKT蛋白表达。结果:ciRS-7在胰腺癌相关成纤维细胞的外泌体中表达上调(P<0.05);CAFs释放的外泌体促进胰腺癌细胞糖酵解和增殖(P<0.05);敲降胰腺癌相关成纤维细胞ciRS-7的表达,其外泌体可抑制胰腺癌细胞糖酵解相关基因的表达(P<0.05);ciRS-7在胰腺癌细胞中经miR-7/IGF1R/AKT通路促进癌细胞糖酵解(P<0.05)。结论:ciRS-7在胰腺癌相关成纤维细胞的外泌体中高表达,下调ciRS-7具有抑制胰腺癌细胞糖酵解的作用。

Objective:To investigate the effect of exosomal ciRS-7 derived from cancer associated fibroblasts(CAFs)on glycolysis of pancreatic cancer cells.Methods:Exosomes derived from CAFs were extracted by ultracentrifugation,and the real-time quantitative polymerase chain reaction(RT-qPCR)was used to detect the expression of exosomal ciRS-7 and the mRNAs of key glycolytic enzymes in pancreatic cancer cells.ciRS-7-WT or ciRS-7-MT was used to verify the relationship between ciRS-7 and miR-7 in pancreatic cancer by Dual Luciferase Reporter Assay.The protein changes of IGF1R/AKT axis in pancreatic cancer were analyzed by Western blot.Results:The expression of ciRS-7 was up-regulated in the exosomes derived from CAFs.The exosomes released by CAFs promoted glycolytic metabolism and proliferation of pancreatic cancer cells.The exosomes released by pancreatic CAFs with ciRS-7 knock-down could inhibit the expression of glycolysis-related genes in pancreatic cancer cells.And ciRS-7 promoted glycolysis in pancreatic cancer cells through modulating the miR-7/IGF1R/AKT pathway.Conclusion:Exosomal ciRS-7 was up-regulated in pancreatic CAFs,and the downregulation of ciRS-7 expression in CAFs could suppress the glycolysis metabolism in pancreatic cancer cells.