Advertisement

Tumor Biology

, Volume 36, Issue 11, pp 8545–8552 | Cite as

Expressions of miR-181a and miR-20a in RPMI8226 cell line and their potential as biomarkers for multiple myeloma

  • Jing Peng
  • Asmitananda Thakur
  • Shuo Zhang
  • Yuanfeng Dong
  • Xiaoqin Wang
  • Ruili Yuan
  • Kaige Zhang
  • Xuan Guo
Research Article

Abstract

Multiple myeloma (MM) is characterized by clonal proliferation of malignant plasma cells in the bone marrow. The anti-tumor activity of bortezomib (a proteosome inhibitor) in MM is challenged by emergence of drug resistance. MicroRNAs (miR) regulate and orchestrate multiple cellular pathways. We investigate the contribution miR-181a and miR-20a expressions’ on cell proliferation and apoptosis in RPMI8226 cell line and their influence on bortezomib treatment. RNA isolation, quantitative real-time PCR (qRT-PCR), cell proliferation assay, cell cycle analysis, and cell apoptosis assay were done. Statistical analysis was performed using SPSS 17.0 software (SPSS, Chicago, IL, USA). P values of less than 0.05 were considered statistically significant. RPMI8226 cells seeded in 96-well plates and treated for 24 h with different concentrations of bortezomib showed dose-dependent growth inhibition; expression of both miR-181a and miR-20a were inhibited by bortezomib. We found decrease of miR-181a (60 %) and miR-20a (30 %) in cells transfected with 20-nM inhibitor. A relative increase of 14-fold in miR-181a and 11-fold in miR-20a was observed in cells transfected with mimics of the same concentration. Transient low expression of miR-181a/20a inhibited proliferation at day 4, and overexpression of miR-181a promoted proliferation. Cells transfected with miR-181a/20a inhibitor within day 4 showed lower survival rate, and low expression of miR-181a on the fourth day after transfection promoted apoptosis. Our findings suggest that miR-181a/20a has a higher expression in MM. miR-181-a expression is proportional to MM tumor burden and could be a biomaker for monitoring treatment. miR-20a shows the potential of a diagnostic biomarker.

Keywords

Multiple myeloma miR-181-a miR-20a Bortezomib Apoptosis Biomarker 

Notes

Acknowledgments

This research was supported by the grant from the Shaanxi Province Science and Technology Research Development Fund (Grant number: 2014 K11-01-02-03).

Conflicts of interest

None

References

  1. 1.
    Kyle RA, Rajkumar SV. Multiple myeloma. Blood. 2008;111:2962–72.CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Mahindra A, Hideshima T, Anderson KC. Multiple myeloma: biology of the disease. Blood Rev. 24 Suppl 1:S5-11.Google Scholar
  3. 3.
    Lawasut P, Groen RW, Dhimolea E, Richardson PG, Anderson KC, Mitsiades CS. Decoding the pathophysiology and the genetics of multiple myeloma to identify new therapeutic targets. Semin Oncol. 40:537–48.Google Scholar
  4. 4.
    Chen CZ, Li L, Lodish HF, Bartel DP. Micrornas modulate hematopoietic lineage differentiation. Science. 2004;303:83–6.CrossRefPubMedGoogle Scholar
  5. 5.
    Dimopoulos K, Gimsing P, Gronbaek K. Aberrant microrna expression in multiple myeloma. Eur J Haematol. 2013;91:95–105.CrossRefPubMedGoogle Scholar
  6. 6.
    Bi CL, Chng WJ. Microrna: important player in the pathobiology of multiple myeloma. Biomed Res Int. 2014.Google Scholar
  7. 7.
    Pichiorri F, Suh SS, Ladetto M, Kuehl M, Palumbo T, Drandi D, et al. Micrornas regulate critical genes associated with multiple myeloma pathogenesis. Proc Natl Acad Sci U S A. 2008;105:12885–90.CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Dimopoulos MA, Delforge M, Hajek R, Kropff M, Petrucci MT, Lewis P, et al. Melphalan, prednisone and lenalidomide followed by lenalidomide maintenance improves health-related quality of life (hrqol), with newly diagnosed multiple myeloma (ndmm) patients ≥65 years benefiting from delays in disease progression. Blood. 2011;118:1365.Google Scholar
  9. 9.
    Zhang WJ, Wang YYE, Zhang Y, Leleu X, Reagan M, Zhang Y, et al. Global epigenetic regulation of micrornas in multiple myeloma. Plos One. 2014;9.Google Scholar
  10. 10.
    Humphries C. Genetics: profiling a shape-shifter. Nature 480:S50-51.Google Scholar
  11. 11.
    Chen L, Li C, Zhang R, Gao X, Qu X, Zhao M, et al. Mir-17-92 cluster micrornas confers tumorigenicity in multiple myeloma. Cancer Lett. 309:62–70.Google Scholar
  12. 12.
    Gao X, Zhang R, Qu X, Zhao M, Zhang S, Wu H, et al. Mir-15a, mir-16-1 and mir-17-92 cluster expression are linked to poor prognosis in multiple myeloma. Leuk Res 36:1505–9.Google Scholar
  13. 13.
    Huang JJ, Yu J, Li JY, Liu YT, Zhong RQ. Circulating microrna expression is associated with genetic subtype and survival of multiple myeloma. Med Oncol. 29:2402–8.Google Scholar
  14. 14.
    Ventura A, Young AG, Winslow MM, Lintault L, Meissner A, Erkeland SJ, et al. Targeted deletion reveals essential and overlapping functions of the miR-17 similar to 92 family of mirna clusters. Cell. 2008;132:875–86.CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Ballabio E, Armesto M, Breeze CE, Manterola L, Arestin M, Tramonti D, et al. Bortezomib action in multiple myeloma: Microrna-mediated synergy (and mir-27a/cdk5 driven sensitivity)? Blood Cancer J. 2:e83.Google Scholar
  16. 16.
    Benevolo G, Larocca A, Gentile M, Pregno P, Gay F, Botto B, et al. The efficacy and safety of bortezomib and dexamethasone as a maintenance therapy in patients with advanced multiple myeloma who are responsive to salvage bortezomib-containing regimens. Cancer Am Cancer Soc. 2011;117:1884–90.Google Scholar
  17. 17.
    Palumbo A, Anderson K. Medical progress multiple myeloma. New Engl J Med. 2011;364:1046–60.CrossRefPubMedGoogle Scholar
  18. 18.
    Lin M, Hou J, Chen W, Huang X, Liu Z, Zhou Y, et al. Improved response rates with bortezomib in relapsed or refractory multiple myeloma: an observational study in Chinese patients. Adv Ther. 31:1082–94.Google Scholar
  19. 19.
    Serrano M. Cancer: a lower bar for senescence. Nature 464:363–4.Google Scholar
  20. 20.
    Jagannathan S, Vad N, Vallabhapurapu S, Anderson KC, Driscoll JJ. Mir-29b replacement inhibits proteasomes and disrupts aggresome+autophagosome formation to enhance the antimyeloma benefit of bortezomib. Leukemia.Google Scholar
  21. 21.
    Kremer-Tal S, Narla G, Chen Y, Hod E, DiFeo A, Yea S, et al. Downregulation of klf6 is an early event in hepatocarcinogenesis, and stimulates proliferation while reducing differentiation. J Hepatol. 2007;46:645–54.CrossRefPubMedGoogle Scholar
  22. 22.
    Palumbo A, Attal M, Roussel M. Shifts in the therapeutic paradigm for patients newly diagnosed with multiple myeloma: maintenance therapy and overall survival. Clin Cancer Res. 2011;17:1253–63.CrossRefPubMedGoogle Scholar
  23. 23.
    Ahmad N, Haider S, Jagannathan S, Anaissie E, Driscoll JJ. Microrna theragnostics for the clinical management of multiple myeloma. Leukemia. 2014;28:732–8.CrossRefPubMedGoogle Scholar
  24. 24.
    Ahmad A, Maitah MY, Ginnebaugh KR, Li YW, Bao B, Gadgeel SM, et al. Inhibition of Hedgehog signaling sensitizes nsclc cells to standard therapies through modulation of emt-regulating mirnas. J Hematol Oncol. 2013;6.Google Scholar
  25. 25.
    Yamamoto N, Kinoshita T, Nohata N, Itesako T, Yoshino H, Enokida H, et al. Tumor suppressive microrna-218 inhibits cancer cell migration and invasion by targeting focal adhesion pathways in cervical squamous cell carcinoma. Int J Oncol. 2013;42:1523–32.PubMedPubMedCentralGoogle Scholar
  26. 26.
    Yamamoto N, Kinoshita T, Nohata N, Yoshino H, Itesako T, Fujimura L, et al. Tumor-suppressive microRNA-29a inhibits cancer cell migration and invasion via targeting hsp47 in cervical squamous cell carcinoma. Int J Oncol. 2013;43:1855–63.PubMedPubMedCentralGoogle Scholar
  27. 27.
    Luo M, Shen DX, Zhou XN, Chen XD, Wang W. Microrna-497 is a potential prognostic marker in human cervical cancer and functions as a tumor suppressor by targeting the insulin-like growth factor 1 receptor. Surgery. 2013;153:836–47.CrossRefPubMedGoogle Scholar
  28. 28.
    Lee YS, Dutta A. The tumor suppressor microrna let-7 represses the hmga2 oncogene. Gene Dev. 2007;21:1025–30.CrossRefPubMedPubMedCentralGoogle Scholar
  29. 29.
    Qu XY, Zhao M, Wu S, Yu WJ, Xu JR, Xu J, et al. Circulating microrna 483-5p as a novel biomarker for diagnosis survival prediction in multiple myeloma. Med Oncol. 2014;31.Google Scholar
  30. 30.
    Lwin T, Lin J, Choi YS, Zhang X, Moscinski LC, Wright KL, et al. Follicular dendritic cell-dependent drug resistance of non-hodgkin lymphoma involves cell adhesion-mediated bim down-regulation through induction of microrna-181a. Blood 116:5228–36.Google Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2015

Authors and Affiliations

  • Jing Peng
    • 1
  • Asmitananda Thakur
    • 2
    • 5
  • Shuo Zhang
    • 2
  • Yuanfeng Dong
    • 3
  • Xiaoqin Wang
    • 4
  • Ruili Yuan
    • 4
  • Kaige Zhang
    • 4
  • Xuan Guo
    • 4
  1. 1.Medical College of Xi’an Jiaotong UniversityXi’an Jiaotong UniversityXi’anChina
  2. 2.Department of Respiratory and Critical Care Medicine, The First Affiliated Hospital, Medical College of Xi’an Jiaotong UniversityXi’an Jiaotong UniversityXi’anChina
  3. 3.Clinical Laboratory of 521 Hospital of the Ordinance IndustryXi’anChina
  4. 4.Department of Clinical Laboratory, The First Affiliated Hospital, Medical College of Xi’an Jiaotong UniversityXi’an Jiaotong UniversityXi’anChina
  5. 5.Department of Internal MedicineLife Guard HospitalBiratnagarNepal

Personalised recommendations