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Exosomal LINC00161 promotes angiogenesis and metastasis via regulating miR-590-3p/ROCK axis in hepatocellular carcinoma

Cancer Gene Therapy volume 28, pages 719–736 (2021)Cite this article

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A Correction to this article was published on 19 April 2021

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Abstract

Hepatocellular carcinoma (HCC) is a lethal malignancy with few effective options for therapeutic treatment in its advanced stages. While exosomal LINC00161 has been identified as a potential biomarker for HCC, its regulatory function and clinical values remain largely unknown. LINC00161 expressions in serum-derived exosomes from HCC patients and HCC cells were determined by qRT-PCR. The ability of proliferation, migration, and angiogenesis in HUVECs was assessed by MTT, Transwell, and tube formation. Luciferase reporter assay and AGO2-RIP assay were conducted to explore the interactions among LINC00161, miR-590-3p, and ROCK2. The level of ROCK signal-related proteins was examined by Western blotting and immunohistochemistry (IHC) assay. Subcutaneous tumor growth was observed in nude mice, in which in vivo metastasis was observed following tail vein injection of HCC cells. High levels of LINC00161 were detected in both serum-derived exosomes from HCC patients and the supernatants of HCC cell lines and were significantly associated with poor survival. Functional study demonstrated that exosomal LINC00161 derived from HCC-cells were significantly associated with enhanced proliferation, migration, and angiogenesis in HUVECs in vitro, all of which were effectively inhibited when LINC00161 was sliced with shRNA in HCC-cells. In vivo experiment showed that LINC00161 loss inhibited tumorigenesis and metastasis of HCC. Mechanistic study revealed that exosome-carried LINC00161 directly targeted miR-590-3p and induced its downstream target ROCK2, finally activating growth/metastasis-related signals in HCC. Exosome-carried LINC00161 promotes HCC tumorigenesis through inhibiting miR-590-3p to activate the ROCK2 signaling pathway, suggesting that LINC00161 may be used as potential targets to improve HCC treatment efficiency.

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Fig. 1: LINC00161 expression was increased in the exosomes of HCC and was associated with poor survival.
Fig. 2: Intercellular transfer of LINC00161 by HCC-derived exosomes to HUVECs promoted cell proliferation, migration, and angiogenesis.
Fig. 3: Exosomes derived from HCC cells with LINC00161 knockdown inhibited HUVECs proliferation, migration, and angiogenesis.
Fig. 4: Exosome-carried LINC00161 bound to miR-590-3p and negatively regulated its expression in HUVECs.
Fig. 5: miR-590-3p directly targeted ROCK2 to inhibit ROCK signaling pathway in HUVECs.
Fig. 6: miR-590-3p inhibited migration and angiogenesis of HUVECs through downregulation of ROCK signaling pathway.
Fig. 7: miR-590-3p/ROCK2 axis mediated the migration and angiogenesis of HUVECs induced by exosome-carried LINC00161. HUVECs were treated with blank, HCCLM3- or MHCC-97H-derived exosomal LINC00161, miR-590 mimics, or Y-27632, respectively.
Fig. 8: Knockdown of LINC00161 inhibited tumorigenesis and metastasis of HCC in vivo.
Fig. 9: Exosome-carried LINC00161 promoted tumorigenesis in vivo.

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References

  1. Balogh J, Victor D III, Asham EH, Burroughs SG, Boktour M, Saharia A, et al. Hepatocellular carcinoma: a review. J Hepatocell Carcinoma. 2016;3:41–53.

    Article  PubMed  PubMed Central  Google Scholar 

  2. Chen W, Zheng R, Baade PD, Zhang S, Zeng H, Bray F, et al. Cancer statistics in China, 2015. CA Cancer J Clin. 2016;66:115–32.

    Article  PubMed  Google Scholar 

  3. Dhir M, Melin AA, Douaiher J, Lin C, Zhen WK, Hussain SM, et al. A review and update of treatment options and controversies in the management of hepatocellular carcinoma. Ann Surg. 2016;263:1112–25.

    Article  PubMed  Google Scholar 

  4. Yu S, Cao H, Shen B, Feng J. Tumor-derived exosomes in cancer progression and treatment failure. Oncotarget. 2015;6:37151–68.

    Article  PubMed  PubMed Central  Google Scholar 

  5. Azmi AS, Bao B, Sarkar FH. Exosomes in cancer development, metastasis, and drug resistance: a comprehensive review. Cancer metastasis Rev. 2013;32:623–42.

    Article  CAS  PubMed  Google Scholar 

  6. Abudoureyimu M, Zhou H, Zhi Y, Wang T, Feng B, Wang R, et al. Recent progress in the emerging role of exosome in hepatocellular carcinoma. Cell Prolif. 2019;52:e12541.

    Article  PubMed  Google Scholar 

  7. Suchorska WM, Lach MS. The role of exosomes in tumor progression and metastasis. Oncol Rep. 2016;35:1237–44.

    Article  CAS  PubMed  Google Scholar 

  8. Zhang H, Chen Z, Wang X, Huang Z, He Z, Chen Y. Long non-coding RNA: a new player in cancer. J Hematol Oncol. 2013;6:37.

    Article  PubMed  PubMed Central  Google Scholar 

  9. Gutschner T, Diederichs S. The hallmarks of cancer: a long non-coding RNA point of view. RNA Biol. 2012;9:703–19.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Zhang C, Yang X, Qi Q, Gao Y, Wei Q, Han S. lncRNA-HEIH in serum and exosomes as a potential biomarker in the HCV-related hepatocellular carcinoma. Cancer biomarkers: Sect A Dis markers. 2018;21:651–9.

    Article  CAS  Google Scholar 

  11. Li X, Lei Y, Wu M, Li N. Regulation of macrophage activation and polarization by HCC-derived exosomal lncRNA TUC339. Int J Mol Sci. 2018;19:2958.

    Article  PubMed Central  Google Scholar 

  12. Xu H, Chen Y, Dong X, Wang X. Serum exosomal long noncoding RNAs ENSG00000258332.1 and LINC00635 for the diagnosis and prognosis of hepatocellular carcinoma. Cancer Epidemiol Biomark Prev. 2018;27:710–6.

    Article  CAS  Google Scholar 

  13. Li B, Mao R, Liu C, Zhang W, Tang Y, Guo Z. LncRNA FAL1 promotes cell proliferation and migration by acting as a CeRNA of miR-1236 in hepatocellular carcinoma cells. Life Sci. 2018;197:122–9.

    Article  CAS  PubMed  Google Scholar 

  14. Xu LC, Chen QN, Liu XQ, Wang XM, Chang QM, Pan Q, et al. Up-regulation of LINC00161 correlates with tumor migration and invasion and poor prognosis of patients with hepatocellular carcinoma. Oncotarget. 2017;8:56168–73.

    Article  PubMed  PubMed Central  Google Scholar 

  15. Sun L, Su Y, Liu X, Xu M, Chen X, Zhu Y, et al. Serum and exosome long non coding RNAs as potential biomarkers for hepatocellular carcinoma. J Cancer. 2018;9:2631–9.

    Article  PubMed  PubMed Central  Google Scholar 

  16. He L, Hannon GJ. MicroRNAs: small RNAs with a big role in gene regulation. Nat Rev Genet. 2004;5:522–31.

    Article  CAS  PubMed  Google Scholar 

  17. Bracken CP, Scott HS, Goodall GJ. A network-biology perspective of microRNA function and dysfunction in cancer. Nat Rev Genet. 2016;17:719–32.

    Article  CAS  PubMed  Google Scholar 

  18. Yao H, Liu N, Lin MC, Zheng J. Positive feedback loop between cancer stem cells and angiogenesis in hepatocellular carcinoma. Cancer Lett. 2016;379:213–9.

    Article  CAS  PubMed  Google Scholar 

  19. Xu X, Tao Y, Shan L, Chen R, Jiang H, Qian Z, et al. The role of microRNAs in hepatocellular carcinoma. J Cancer. 2018;9:3557–69.

    Article  PubMed  PubMed Central  Google Scholar 

  20. Salem M, O’Brien JA, Bernaudo S, Shawer H, Ye G, Brkic J, et al. miR-590-3p promotes ovarian cancer growth and metastasis via a novel FOXA2-versican pathway. Cancer Res. 2018;78:4175–90.

    Article  CAS  PubMed  Google Scholar 

  21. Di Agostino S, Valenti F, Sacconi A, Fontemaggi G, Pallocca M, Pulito C, et al. Long non-coding MIR205HG depletes Hsa-miR-590-3p leading to unrestrained proliferation in head and neck squamous cell carcinoma. Theranostics. 2018;8:1850–68.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Zhang J, Jin M, Chen X, Zhang R, Huang Y, Liu H, et al. Loss of PPM1F expression predicts tumour recurrence and is negatively regulated by miR-590-3p in gastric cancer. Cell Prolif. 2018;51:e12444.

    Article  PubMed  PubMed Central  Google Scholar 

  23. He B, Lu P, Guan L, Li T, Zhang L, Zhu QG, et al. Identifying key regulating miRNAs in hepatocellular carcinomas by an omics’ method. Oncotarget. 2017;8:103919–30.

    Article  PubMed  PubMed Central  Google Scholar 

  24. Li TM, Liu SC, Huang YH, Huang CC, Hsu CJ, Tsai CH, et al. YKL-40-Induced inhibition of miR-590-3p promotes interleukin-18 expression and angiogenesis of endothelial progenitor cells. int. J. Mol. Sci. 2017;18:920.

  25. Bao MH, Li GY, Huang XS, Tang L, Dong LP, Li JM. Long Noncoding RNA LINC00657 Acting as a miR-590-3p Sponge to Facilitate Low Concentration Oxidized Low-Density Lipoprotein-Induced Angiogenesis. Mol Pharm. 2018;93:368–75.

    Article  CAS  Google Scholar 

  26. Zhang JG, Zhang DD, Liu Y, Hu JN, Zhang X, Li L, et al. RhoC/ROCK2 promotes vasculogenic mimicry formation primarily through ERK/MMPs in hepatocellular carcinoma. Biochim Biophys Acta Mol Basis Dis. 2019;1865:1113–25.

    Article  CAS  PubMed  Google Scholar 

  27. Wong CC, Wong CM, Tung EK, Man K, Ng IO. Rho-kinase 2 is frequently overexpressed in hepatocellular carcinoma and involved in tumor invasion. Hepatology. 2009;49:1583–94.

    Article  CAS  PubMed  Google Scholar 

  28. Liu J, Wada Y, Katsura M, Tozawa H, Erwin N, Kapron CM, et al. Rho-associated coiled-coil kinase (ROCK) in molecular regulation of angiogenesis. Theranostics. 2018;8:6053–69.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Castellano JJ, Navarro A, Vinolas N, Marrades RM, Moises J, Cordeiro A, et al. LincRNA-p21 impacts prognosis in resected non-small cell lung cancer patients through angiogenesis regulation. J Thorac Oncol. 2016;11:2173–82.

    Article  PubMed  Google Scholar 

  30. Zhao X, Liu Y, Li Z, Zheng S, Wang Z, Li W, et al. Linc00511 acts as a competing endogenous RNA to regulate VEGFA expression through sponging hsa-miR-29b-3p in pancreatic ductal adenocarcinoma. J Cell Mol Med. 2018;22:655–67.

    Article  CAS  PubMed  Google Scholar 

  31. Muller S, Raulefs S, Bruns P, Afonso-Grunz F, Plotner A, Thermann R, et al. Next-generation sequencing reveals novel differentially regulated mRNAs, lncRNAs, miRNAs, sdRNAs and a piRNA in pancreatic cancer. Mol Cancer. 2015;14:94.

    Article  PubMed  PubMed Central  Google Scholar 

  32. Tsai MC, Spitale RC, Chang HY. Long intergenic noncoding RNAs: new links in cancer progression. Cancer Res. 2011;71:3–7.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Bryan BA, Dennstedt E, Mitchell DC, Walshe TE, Noma K, Loureiro R, et al. RhoA/ROCK signaling is essential for multiple aspects of VEGF-mediated angiogenesis. FASEB J. 2010;24:3186–95.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Wong CC, Wong CM, Tung EK, Au SL, Lee JM, Poon RT, et al. The microRNA miR-139 suppresses metastasis and progression of hepatocellular carcinoma by down-regulating Rho-kinase 2. Gastroenterology. 2011;140:322–31.

    Article  CAS  PubMed  Google Scholar 

  35. Xiang Q, Chen W, Ren M, Wang J, Zhang H, Deng DY, et al. Cabozantinib suppresses tumor growth and metastasis in hepatocellular carcinoma by a dual blockade of VEGFR2 and MET. Clin Cancer Res. 2014;20:2959–70.

    Article  CAS  PubMed  Google Scholar 

  36. Gramantieri L, Baglioni M, Fornari F, Laginestra MA, Ferracin M, Indio V, et al. LncRNAs as novel players in hepatocellular carcinoma recurrence. Oncotarget. 2018;9:35085–99.

    Article  PubMed  PubMed Central  Google Scholar 

  37. Wang Y, Zhang L, Zheng X, Zhong W, Tian X, Yin B, et al. Long non-coding RNA LINC00161 sensitises osteosarcoma cells to cisplatin-induced apoptosis by regulating the miR-645-IFIT2 axis. Cancer Lett. 2016;382:137–46.

    Article  CAS  PubMed  Google Scholar 

  38. Xu M, Zhou K, Wu Y, Wang L, Lu S. Linc00161 regulated the drug resistance of ovarian cancer by sponging microRNA-128 and modulating MAPK1. Mol Carcinog. 2019;58:577–87.

    Article  CAS  PubMed  Google Scholar 

  39. Conigliaro A, Costa V, Lo Dico A, Saieva L, Buccheri S, Dieli F, et al. CD90+ liver cancer cells modulate endothelial cell phenotype through the release of exosomes containing H19 lncRNA. Mol Cancer. 2015;14:155.

    Article  PubMed  PubMed Central  Google Scholar 

  40. Li R, Wang Y, Zhang X, Feng M, Ma J, Li J, et al. Exosome-mediated secretion of LOXL4 promotes hepatocellular carcinoma cell invasion and metastasis. Mol Cancer. 2019;18:18.

    Article  PubMed  PubMed Central  Google Scholar 

  41. He M, Qin H, Poon TC, Sze SC, Ding X, Co NN, et al. Hepatocellular carcinoma-derived exosomes promote motility of immortalized hepatocyte through transfer of oncogenic proteins and RNAs. Carcinogenesis. 2015;36:1008–18.

    Article  CAS  PubMed  Google Scholar 

  42. Shan Y, Ma J, Pan Y, Hu J, Liu B, Jia L. LncRNA SNHG7 sponges miR-216b to promote proliferation and liver metastasis of colorectal cancer through upregulating GALNT1. Cell Death Dis. 2018;9:722.

    Article  PubMed  PubMed Central  Google Scholar 

  43. Miao L, Liu HY, Zhou C, He X. LINC00612 enhances the proliferation and invasion ability of bladder cancer cells as ceRNA by sponging miR-590 to elevate expression of PHF14. J Exp Clin Cancer Res. 2019;38:143.

    Article  PubMed  PubMed Central  Google Scholar 

  44. Ueno K, Hirata H, Shahryari V, Chen Y, Zaman MS, Singh K, et al. Tumour suppressor microRNA-584 directly targets oncogene Rock-1 and decreases invasion ability in human clear cell renal cell carcinoma. Br J Cancer. 2011;104:308–15.

    Article  CAS  PubMed  Google Scholar 

  45. Hoang MV, Whelan MC, Senger DR. Rho activity critically and selectively regulates endothelial cell organization during angiogenesis. Proc Natl Acad Sci USA. 2004;101:1874–9.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  46. Lin C, Song L, Liu A, Gong H, Lin X, Wu J, et al. Overexpression of AKIP1 promotes angiogenesis and lymphangiogenesis in human esophageal squamous cell carcinoma. Oncogene. 2015;34:384–93.

    Article  CAS  PubMed  Google Scholar 

  47. Kessenbrock K, Plaks V, Werb Z. Matrix metalloproteinases: regulators of the tumor microenvironment. Cell. 2010;141:52–67.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Funding

This work was supported by Scientific Research and Innovation Fund of Xinjiang Medical University (NO.XJC2013118).

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Author notes

  1. These authors contributed equally: Li-Na You, Qin-Wen Tai, Lin Xu

Authors and Affiliations

  1. Department of Traditional Chinese Medicine, the Fifth Affiliated Hospital of Xinjiang Medical University, Urumqi, 830011, PR China

    Li-Na You

  2. Traditional Chinese Medicine Gynecology, Maternal and Child Health Hospital of Xinjiang Uyghur Autonomous Region, Urumqi, 830011, PR China

    Li-Na You

  3. Department of General Surgery, ShenZhen Hospital of Southern Medical University, Shenzhen, 518000, PR China

    Qin-Wen Tai, Heng Zhang & Wu-Kui Huang

  4. Department of Hepatobiliary& Pancreatic Surgery, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital & Shenzhen Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Shenzhen, 518000, PR China

    Lin Xu

  5. Department of Ultrasound Medicine, ShenZhen Hospital of Southern Medical University, Shenzhen, 518000, PR China

    Yi Hao

  6. Cancer Institute, Cancer Hospital Affiliated to Xinjiang Medical University, Urumqi, 830011, PR China

    Wen-Jia Guo

  7. Department of Pulmonary Medicine, Cancer Hospital Affiliated to Xinjiang Medical University, Urumqi, 830011, PR China

    Qiao Zhang

  8. Department of Hepatobiliary and Pancreatic Surgery, Cancer Hospital Affiliated to Xinjiang Medical University, Urumqi, 830011, PR China

    Qing Tong

  9. InterventionaL Diagnosis and Treatment Department, Affiliated Tumor Hospital of Xinjiang Medical University, Urumqi, 830011, PR China

    Wu-Kui Huang

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Contributions

Guarantor of integrity of the entire study: Wu-Hui Huang; study concepts: Wu-Hui Huang, Qin-Wen Tai; study design: Wu-Hui Huang, Li-Na You, Lin Xu, Yi Hao; definition of intellectual content: Lin Xu; literature research: Li-Na You; clinical studies: Wen-JiaGuo; experimental studies: Wu-Hui Huang, Wen-JiaGuo; data acquisition: Qiao Zhang; data analysis: Wu-Hui Huang, Qing Tong; statistical analysis: Qiao Zhan, Li-Na You; manuscript preparation: Wu-Hui Huang, Heng Zhang; manuscript editing: Qing Tong; manuscript review: Heng Zhang.

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Correspondence to Wu-Kui Huang.

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The study protocol was approved by the Medical Ethics Committee of Shenzhen Hospital of Southern Medical University and all patients provided written informed consent. All experimentations involving animals were carried out in full accordance with the institutional animal welfare guideline and approved by Shenzhen Hospital of Southern Medical University.

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You, LN., Tai, QW., Xu, L. et al. Exosomal LINC00161 promotes angiogenesis and metastasis via regulating miR-590-3p/ROCK axis in hepatocellular carcinoma. Cancer Gene Ther 28, 719–736 (2021). https://doi.org/10.1038/s41417-020-00269-2

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