| Gastric cancer(GC)is a global health problem,the third leading cause of cancerrelated deaths and the fifth most diagnosed malignancy worldwide.Despite recent advances in treatment,the prognosis for patients with advanced GC remains poor,with an overall 5-year survival rate of approximately 25%,mainly due to frequent recurrence,metastasis and drug resistance.Exosomes are 40-150 nm vesicles synthesized in the endosomal pathway of normal and tumor cells.They consist of lipid bilayers that carry miRNA and mRNA,cytokines,chemokines,growth factors,etc.within them.The communication between cells within the Tumor microenvironment(TME),whether paracrine,autocrine or even endocrine,cannot be separated from the communication role of exosomes in it.Currently,there are more studies on RNA of exosomal vesicle contents and less studies on protein level,so we will start our study from protein perspective.In this study,we first compared the differences in metastatic ability of different gastric cancer cell lines by migration invasion,proliferation assay and EMT index detection.We then extracted exosomes from high metastatic gastric cancer cells to treat low gastric cancer cells and found that their migration invasion,clone formation,proliferation ability,and EMT were all improved compared with the untreated group;by reviewing the literature and exosome database,we selected ENO1,a vesicle content,as the target of our study.We extracted protein and RNA to verify the expression content of ENO1 in different gastric cancer cells,and found that the high expression of ENO1 molecules in high metastatic gastric cancer cell lines was consistent with our above view,so we launched further studies.We established high metastatic cells stably overexpressing and knocking down ENO1 molecules,extracted exosomes of overexpressing ENO1 stably transduced cell lines,and identified them by transmission electron microscopy,nanoparticle tracking assay,and protein blotting assay,while exosomes of overexpressing ENO1 gastric cancer cell lines were used as stimulating factors to treat gastric cancer parental cells,compared with the group without exosomes and adding exosome inhibitor The migratory invasion ability was significantly enhanced in the group with the addition of overexpressed ENO1 exosomes compared to the GW4869 group.It may also activate PI3K/Akt signaling pathway to promote EMT.High levels of ENO1,a non-rate-limiting enzyme of the glycolytic pathway,can independently regulate the level of glycolysis in gastric cancer cells.We examined the extracellular ATP production,glucose consumption and lactate production in overexpressed and knockdown gastric cancer cell lines,and the results were the same as the in vitro functional assays described above.To further develop the discussion,we treated parental cells with the glycolysis inhibitor 2-DG and found that their migratory invasion ability was reduced;glycolysis was inhibited.2-DG treatment of ENO1-stable cell lines resulted in a decrease in migratory invasion ability in both the overexpressed ENO1 group and control cells,but the difference between them was significantly reduced compared to the untreated group,suggesting that 2-DG was able to reply to the ENO1 induced changes in the migratory invasion ability of gastric cancer cells.Therefore,we conclude that ENO1 promotes the epithelial-mesenchymal transition and migration-invasive ability of gastric cancer cells by increasing the glycolytic activation of PI3K/Akt signaling pathway.Through the above experiments we confirmed that high expression of ENO1-derived exosomes promoted gastric cancer cell metastasis by directly enhancing the glycolytic pathway.Liver cancer is one of the causes of cancer deaths worldwide and is a major challenge in global health care.Treatment of hepatocellular carcinoma depends on the size of the tumor,its location and liver function.Treatment measures include surgical resection,local ablation,transarterial chemoembolization and liver transplantation.Surgical resection remains the most effective treatment for early-stage liver cancer,but due to the insidious nature of early-stage liver cancer,many patients are usually diagnosed in the middle to late stages.Meanwhile,the 5-year survival rate of liver cancer patients remains low.Exosomes are extracellular vesicles 40-150 nm in diameter secreted by fusion of multivesicular bodies(MVB)with the plasma membrane,which can be secreted by many types of cells and are widely present in body fluids.Their contents,including nucleic acids,proteins and lipids,are important mediators of complex crosstalk in intercellular interactions.There is growing evidence that tumor cells can alter the tumor microenvironment by secreting EVs,which play an important role in tumorigenesis development.In this study,we demonstrate that heterogeneity between hepatocellular carcinoma cells with different metastatic potential is conveyed through exosomal communication.We previously found that the conditioned medium of high metastatic hepatocellular carcinoma cells promoted the migratory invasive ability and self-renewal ability of low metastatic hepatocellular carcinoma cells,and the promotion effect was diminished by removing the conditioned medium treatment with exosomes,so we investigated exosomes as a window for targeting heterogeneity.We first extracted exosomes from high metastatic potential hepatocellular carcinoma cells and identified them by particle size analysis,electron microscopy and protein blotting;meanwhile,immunofluorescence experiments demonstrated that exosomes from high metastatic potential hepatocellular carcinoma cells could be endocytosed by low metastatic hepatocellular carcinoma cells.We extracted high metastatic potential hepatocellular carcinoma cell exosomes to treat low metastatic hepatocellular carcinoma cells,and the results showed that exosomes promoted the migration invasion,self-renewal,proliferation ability and epithelial-mesenchymal transition phenotype of hepatocellular carcinoma cells.PI3K/Akt/mTOR signaling pathway to promote a range of malignant phenotypes. |
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