Cholangiocarcinoma derived exosomes break down vascular integrity and cause metastasis by triggering endoplasmic reticulum stress in vascular endothelial cells
LI Zhengzheng1, SUN Hongwei2, ZHANG Nan1, SU Huafang3
1.Department of Neurology, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325015, China; 2.Department of Hepatobiliary Surgery, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325015, China; 3.Department of Chemotherapy, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325015, China
LI Zhengzheng,SUN Hongwei,ZHANG Nan, et al. Cholangiocarcinoma derived exosomes break down vascular integrity and cause metastasis by triggering endoplasmic reticulum stress in vascular endothelial cells[J]. JOURNAL OF WEZHOU MEDICAL UNIVERSITY, 2019, 49(4): 235-242.
Abstract：Objective: To explore the effects of cholangiocarcinoma exosomes on the endothelial tight junction and the possible underlying mechanism. Methods: Exosomes were isolated from the supernatant of cholangiocarcinoma cell line CCLP and human intrahepatic bile duct epithelial cells (HIBEC) by ultracentrifugation according to the well-established protocol and were administrated to human umbilical vein endothelial cells (HUVEC), PBS was chosen as control. Endothelial monolayer permeability was assessed in vitro and tight junction proteins (TJPs) were evaluated by Western blot and immunofluorescence. BALB/c null mice were divided as three groups (PBS, exoCCLP and exoHIBEC), which were administrated with PBS, CCLP exosomes or HIBEC exosomes via tail vein respectively for indicated durations. FITC-Dextran or CCLP were then injected intravenously and leakage of FITC-Dextran in lung and liver, or pulmonary metastases were assessed. The endoplasmic reticulum stress related genes and proteins were analyzed by real-time PCR and Western blot. Endothelial PERK, ATF6 or IRE1α were partially knocked down by siRNA before HUVECs were treated with CCLP exosomes, and TJPs were evaluated afterwards. Results: CCLP exosomes downregulated the expression of TJPs in HUVECs, and increased the permeability of endothelial monolayer in vitro. In vivo experiments indicated that compared to
[1] LIU Y, GU Y, HAN Y, et al. Tumor exosomal RNAs promote lung pre-metastatic niche formation by activating alveolar epithelial TLR3 to recruit neutrophils[J]. Cancer Cell, 2016, 30(2): 243-256.
[2] VALADI H, EKSTR M K, BOSSIOS A, et al. Exosome-mediated transfer of mRNAs and microRNAs is a novel mechanism of genetic exchange between cells[J]. Nat Cell Biol, 2007, 9(6): 654-659.
[3] KANG K, MA R, CAI W, et al. Exosomes secreted from CXCR4 overexpressing mesenchymal stem cells promote cardioprotection via Akt signaling pathway following myocardial infarction[J]. Stem Cells Int, 2015: 659890.
[4] RUIVO C, ADEM B, SILVA M, et al. The biology of cancer exosomes: insights and new perspectives[J]. Cancer Res, 2017, 77(23): 6480-6488.
[5] WEBBER J, STEADMAN R, MASON M D, et al. Cancer exosomes trigger fibroblast to myofibroblast differentiation [J]. Cancer Res, 2010, 70(23): 9621-9630.
[6] LIU Y, GU Y, CAO X. The exosomes in tumor immunity[J]. Oncoimmunology, 2015, 4(9): e1027472.
[7] SANCHEZ C A, ANDAHUR E I, VALENZUELA R, et al. Exosomes from bulk and stem cells from human prostate cancer have a differential microRNA content that contributes cooperatively over local and pre-metastatic niche[J]. Oncotarget, 2016, 7(4): 3993-4008.
[8] UMEZU T, OHYASHIKI K, KURODA M, et al. Leukemia cell to endothelial cell communication via exosomal miRNAs[J]. Oncogene, 2016, 32(22): 2747-2755.
[9] GAJOS-MICHNIEWICZ A, DUECHLER M, CZYZ M. MiRNA in melanoma-derived exosomes[J]. Cancer Lett, 2014, 347(1): 29-37.
[10] STRILIC B, YANG L, ALBARRÁN-JUÁREZ J, et al. Tumour-cell-induced endothelial cell necroptosis via death receptor 6 promotes metastasis[J]. Nature, 2016, 536(7615): 215-218.
[11] ZHOU W, FONG M Y, MIN Y, et al. Cancer-secreted miR-105 destroys vascular endothelial barriers to promote metas-tasis[J]. Cancer Cell, 2014, 25(4): 501-515.
[12] TADOKORO H, UMEZU T, OHYASHIKI K, et al. Exosomes derived from hypoxic leukemia cells enhance tube formation in endothelial cells[J]. J Biol Chem, 2013, 288 (48): 34343-34351.
[13] HONG B S, CHO J H, KIM H, et al. Colorectal cancer cell-derived microvesicles are enriched in cell cycle-related mRNAs that promote proliferation of endothelial cells[J]. BMC Genomics, 2009, 10(1): 556.
[14] CHAN Y K, ZHANG H, LIU P, et al. Proteomic analysis of exosomes from nasopharyngeal carcinoma cell identifies intercellular transfer of angiogenic proteins[J]. Int J Cancer, 2015, 137(8): 1830-1841.
[15] HSU Y L, HUNG J Y, CHANG W A, et al. Hypoxic lung cancer-secreted exosomal miR-23a increased angiogenesis and vascular permeability by targeting prolyl hydroxylase and tight junction protein ZO-1[J]. Oncogene, 2017, 36(34): 4929-4942.
[16] LI J, LI Z, JIANG P, et al. Circular RNA IARS (circ-IARS) secreted by pancreatic cancer cells and located within exosomes regulates endothelial monolayer permeability to promote tumor metastasis[J]. J Exp Clin Cancer Res, 2018, 37(1): 177.
[17] REYMOND N, D'AGUA B B, RIDLEY A J. Crossing the endothelial barrier during metastasis[J]. Nat Rev Cancer, 2013, 13(12): 858-870.
[18] HOTAMISLIGIL G S. Endoplasmic reticulum stress and the inflammatory basis of metabolic disease[J]. Cell, 2010, 140(6): 900-917.
[19] HETZ C. The unfolded protein response: controlling cell fate decisions under ER stress and beyond[J]. Nat Rev Mol Cell Biol, 2013, 13(2): 89-102.
[20] WALTER P, RON D. The unfolded protein response: from stress pathway to homeostatic regulation[J]. Science, 2011, 334(6059): 1081-1086.
[21] WANG S, KAUFMAN R J. The impact of the unfolded protein response on human disease[J]. J Cell Biol, 2012, 197(7): 857-867.
[22] LIANG G, YANG J, WANG Z, et al. Polycystin-2 down-regulates cell proliferation via promoting PERK-dependent phosphorylation of eIF2alpha[J]. Hum Mol Genet, 2008, 17(20): 3254-3262.
[23] POMMIER A, ANAPARTHY N, MEMOS N, et al. Unresolved endoplasmic reticulum stress engenders immune-resistant, latent pancreatic cancer metastases[J]. Science, 2018, 360(6394). pii: eaao4908.
[24] VOLMER R, VAN DER PLOEG K, RON D. Membrane lipid saturation activates endoplasmic reticulum unfolded protein response transducers through their transmembrane do-mains[J]. Proc Natl Acad Sci U S A, 2013, 110(12): 4628-4633.
