Exosome-derived microRNA-29c Induces Apoptosis of BIU-87 Cells by Down Regulating BCL-2 and MCL-1

  • Xu, Xiang-Dong (Department of Urology, The First Affiliated Hospital of Chongqing Medical University) ;
  • Wu, Xiao-Hou (Department of Urology, The First Affiliated Hospital of Chongqing Medical University) ;
  • Fan, Yan-Ru (College of Laboratory Medicine of Chongqing Medical University) ;
  • Tan, Bing (Department of Urology, The First Affiliated Hospital of Chongqing Medical University) ;
  • Quan, Zhen (Department of Urology, The First Affiliated Hospital of Chongqing Medical University) ;
  • Luo, Chun-Li (College of Laboratory Medicine of Chongqing Medical University)
  • Published : 2014.04.30


Background: Aberrant expression of the microRNA-29 family is associated with tumorigenesis and cancer progression. As transport carriers, tumor-derived exosomes are released into the extracellular space and regulate multiple functions of target cells. Thus, we assessed the possibility that exosomes could transport microRNA-29c as a carrier and correlations between microRNA-29c and apoptosis of bladder cancer cells. Materials and Methods: A total of 28 cancer and adjacent tissues were examined by immunohistochemistry to detect BCL-2 and MCL-1 expression. Disease was Ta-T1 in 12 patients, T2-T4 in 16, grade 1 in 8, 2 in 8 and 3 in 12. The expression of microRNA-29c in cancer tissues was detected by quantitative reverse transcriptase PCR (QRT-PCR). An adenovirus containing microRNA-29c was used to infect the BIU-87 human bladder cancer cell line. MicroRNA-29c in exosomes was measured by QRT-PCR. After BIU-87 cells were induced by exosomes-derived microRNA-29c, QRT-PCR was used to detect the level of microRNA-29c. Apoptosis was examined by flow cytometry and BCL-2 and MCL-1 mRNA expressions were assessed by reverse transcription-polymerase chain reaction. Western blotting was used to determine the protein expression of BCL-2 and MCL-1. Results: The expressions of BCL-2 and MCL-1 protein were remarkably increased in bladder carcinoma (p<0.05), but was found mainly in the basal and suprabasal layers in adjacent tissues. The expression of microRNA-29c in cancer tissues was negatively correlated with the BCL-2 and MCL-1. The expression level of microRNA-29c in exosomes and BIU-87 cells from the experiment group was higher than that in control groups (p<0.05). Exosome-derived microRNA-29c induced apoptosis (p<0.01). Although only BCL-2 was reduced at the mRNA level, both BCL-2 and MCL-1 were reduced at the protein level. Conclusions: Human bladder cancer cells infected by microRNA-29c adenovirus can transport microRNA-29c via exosomes. Moreover, exosome-derived microRNA29c induces apoptosis in bladder cancer cells by down-regulating BCL-2 and MCL-1.


  1. Wang Y, Zhang X, Li H, et al (2013). The role of miRNA-29 family in cancer. European J Cell Biol, 92, 123-8.
  2. Mittelbrunn M, Gutiérrez-Vázquez C, Villarroya-Beltri C, et al (2011). Unidirectional transfer of microRNA-loaded exosomes from T cells to antigen-presenting cells. Nat Commun, 2, 282.
  3. Park SY, Lee JH, Ha M, et al (2008). miR-29 miRNAs activate p53 by targeting p85 and CDC42. Nature Structural & Molecular Biology, 16, 23-9.
  4. Reid G, Kirschner MB, van Zandwijk N (2011). Review Circulating microRNAs: Association with disease and potential use as biomarkers. Crit Rev Oncol Hematol, 80, 193-208.
  5. Wang G, Zhang Hh, He Hd, et al (2010). Up-regulation of microRNA in bladder tumor tissue is not common. Int Urol Nephrol, 42, 95-102
  6. Xiong Y, Fang JH, Yun JP, et al (2010). Effects of microRNA-29 on apoptosis, tumorigenicity, and prognosis of hepatocellular carcinoma. Hepatology, 51, 836-45.
  7. Zhang JX, Qian D, Wang FW, et al (2013). MicroRNA-29c enhances the sensitivities of human nasopharyngeal carcinoma to cisplatin-based chemotherapy and radiotherapy. Cancer Letters, 329, 91-8.
  8. Zhao BS, Liu SG, Wang TY, et al (2013). Screening of microRNA in patients with esophageal cancer at same tumor node metastasis stage with different prognoses. Asian Pac J Cancer Prev, 14, 139-43.
  9. Garzon R , Liu S , Fabbri M, et al (2009). MicroRNA-29b induces global DNA hypomethylation and tumor suppressor gene reexpression in acute myeloid leukemia by targeting directly DNMT3A and 3B and indirectly DNMT1. Blood, 113, 6411-8.
  10. Friedman JM, Liang G, Liu CC, et al (2009). The putative tumor suppressor microRNA-101 modulates the cancer epigenome by repressing the polycomb group protein EZH2. Cancer Res, 69, 2623-9.
  11. Fan W, Tian XD, Huang E, et al (2013). Exosomes from CIITAtransfected CT26 cells enhance anti- tumor effects. Asian Pac J Cancer Prev, 14, 987-91.
  12. Garzon R, Heaphy CE, Havelange V, et al (2009), MicroRNA 29b functions in acute myeloid leukemia. Blood, 219, 214-21.
  13. Hu G, Yao H, Chaudhuri AD, et al (2012). Exosome-mediated shuttling of microRNA-29 regulates HIV Tat and morphinemediated neuronal dysfunction. Cell Death Dis, 3, 381.
  14. Jamiyandorj U, Bae JS, Noh SJ, et al (2013). Expression of peptidyl-prolyl isomerase PIN1 and its role in the pathogenesis of extrahepatic cholangiocarcinoma. Oncol Lett November, 6, 1421-6.
  15. Kozomara A, Griffiths-Jones S (2011). miRBase: integrating microRNA annotation and deep-sequencing data. Nucleic Acids Re, 39, 152-7.
  16. Kosaka N, Iguchi H, Yoshioka Y, et al (2010). Secretory mechanisms and intercellular transfer of microRNAs in living cells. J Biol Chem , 285, 17442-52.
  17. Liu YQ, Li Y, Qin J, et al (2014). Matrine reduces proliferation of human lung cancer cells by inducing apoptosis and changing miRNA expression profiles. Asian Pac J Cancer Prev, 15, 2169-77.
  18. Mott JL, Kobayashi S, Bronk SF, et al (2007). Mir-29 regulates Mcl-1 protein expression and apoptosis. Oncogene, 26, 6133-40.
  19. Alvarez-Erviti L, Seow Y, Yin HF, et al (2011). Delivery of siRNA to the mouse brain by systemic injection of targeted exosomes. Nat Biotechnol, 29, 341-5.
  20. Baffa R, Fassan M, Volinia S, et al (2009). MicroRNA expression profiling of human metastatic cancers identifies cancer gene targets. J Pathol, 219, 214-21.
  21. Calin GA, Ferracin M, Cimmino A, et al (2005). A MicroRNA signature associated with prognosis and progression in chronic lymphocytic leukemia. N Engl J Med, 353, 1793-801.
  22. Castilla MA, Moreno-Bueno G, Romero-Perez L, et al (2011). Micro-RNA signature of the epithelial-mesenchymal transition in endometrial carcinosarcoma. J Pathol, 223, 72-80.
  23. Chaput N, Schartz NE, Andre F, et al (2004). Exosomes as potent cell-free peptide-based vaccine. II. Exosomes in CpG adjuvants efficiently prime naive Tc1 lymphocytes leading to tumor rejection. J Immunol, 172, 2137-46.
  24. Ciesla M, Skrzypek K, Kozakowska M, et al (2011). Review MicroRNAs as biomarkers of disease onset. Anal Bioanal Chem, 401, 2051-61.
  25. Dyrskjøt L, Ostenfeld MS, Bramsen JB, et al (2009). Genomic profiling of microRNAs in bladder cancer: miR-129 is associated with poor outcome and promotes cell death in vitro. Cancer Res, 69, 4851-60.

Cited by

  1. MicroRNA-328 Inhibits Proliferation of Human Melanoma Cells by Targeting TGFB2 vol.16, pp.4, 2015,
  2. MicroRNA-29s could target AKT2 to inhibit gastric cancer cells invasion ability vol.32, pp.1, 2015,
  3. Understanding the Role of Non-Coding RNAs in Bladder Cancer: From Dark Matter to Valuable Therapeutic Targets vol.18, pp.7, 2017,
  4. Bortezomib-induced miRNAs direct epigenetic silencing of locus genes and trigger apoptosis in leukemia vol.8, pp.11, 2017,
  5. Effect of exosomal miRNA on cancer biology and clinical applications vol.17, pp.1, 2018,
  6. MicroRNAs in Smoking-Related Carcinogenesis: Biomarkers, Functions, and Therapy vol.7, pp.5, 2018,