The Effect of Long Non-Coding RNA PVT1 Inhibition by Antisense LNA GapmeRs Technology on Proliferation of Human Acute Erythroleukemia Cells

Document Type : Original Article (s)

Authors

1 MSc Student, Department of Genetics and Molecular Biology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

2 Associate Professor, Department of Genetics and Molecular Biology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

Abstract

Background: Long non-coding RNAs (LncRNAs) are a class of RNA molecules with 200-10000 nucleotides in length that have lost their protein coding capacity. They play important role in numerous biological processes including cell proliferation, differentiation, apoptosis, and cancer development. Acute myeloid leukemia (AML) is the most common leukemia. Evidence has shown that long non-coding RNAs PVT1 is upregulated in acute myeloid leukemia, and leads to an increase in cell proliferation. This study aimed to assess the effect of long non-coding RNA PVT1 inhibition by antisense LNA GapmeRs technology on proliferation of human acute erythroleukemia cells.Methods: Human acute erythroleukemia cells (KG1), known as subgroup six of acute myeloid leukemia based on the French–American–British (FAB) classification, were cultured in Roswell Park Memorial Institute (RPMI) medium. Long non-coding RNA PVT1 degradation was performed using antisense LNA GapmeRs technology. At different time points after transfection (24, 48, and 72 hours), long non-coding RNA PVT1 expression and cell proliferation were assessed. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was accomplished to evaluate the long non-coding RNAPVT1 expression. Moreover, Cell viability was measured using 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyl tetrazolium bromide (MTT) assay.Findings: Long non-coding RNA PVT1 expression decreased at 24, 48, and 72 hours after transfection in the antisense LNA GapmeRs group compared to control groups. Cell viability and proliferation was significantly decreased in antisense LNA GapmeRs transfected group compared with other two groups.Conclusion: The present survey exhibited that inhibition of long non-coding PVT1 by using antisense LNA GapmeRs can dramatically reduce the proliferation of human acute erythroleukemia cells. Our results can be helpful in translational medicine for antisense therapy in acute myeloid leukemia.

Keywords

References

  1. Salehi M, Sharifi M. Exosomal miRNAs as novel cancer biomarkers: Challenges and opportunities. J Cell Physiol 2018; 233(9): 6370-80.
  2. Bolha L, Ravnik-Glavac M, Glavac D. Long noncoding RNAs as biomarkers in cancer. Dis Markers 2017; 2017: 7243968.
  3. Gibb EA, Brown CJ, Lam WL. The functional role of long non-coding RNA in human carcinomas. Mol Cancer 2011; 10: 38.
  4. 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.
  5. Du Z, Fei T, Verhaak RG, Su Z, Zhang Y, Brown M, et al. Integrative genomic analyses reveal clinically relevant long noncoding RNAs in human cancer. Nat Struct Mol Biol 2013; 20(7): 908-13.
  6. Colombo T, Farina L, Macino G, Paci P. PVT1: A rising star among oncogenic long noncoding RNAs. Biomed Res Int 2015; 2015: 304208.
  7. Hauptman N, Glavac D. Long non-coding RNA in cancer. Int J Mol Sci 2013; 14(3): 4655-69.
  8. Zand AM, Imani S, Saadati M, Borna H, Ziaei R, Honari H. Effect of age, gender and blood group on blood cancer types. Kowsar Medical Journal 2010; 15(2): 111-4. [In Persian].
  9. Lowenberg B, Downing JR, Burnett A. Acute myeloid leukemia. N Engl J Med 1999; 341(14): 1051-62.
  10. Bennett JM, Catovsky D, Daniel MT, Flandrin G, Galton DA, Gralnick HR, et al. Proposed revised criteria for the classification of acute myeloid leukemia. A report of the French-American-British Cooperative Group. Ann Intern Med 1985; 103(4): 620-5.
  11. Hasserjian RP, Zuo Z, Garcia C, Tang G, Kasyan A, Luthra R, et al. Acute erythroid leukemia: A reassessment using criteria refined in the 2008 WHO classification. Blood 2010; 115(10): 1985-92.
  12. Zuo Z, Polski JM, Kasyan A, Medeiros LJ. Acute erythroid leukemia. Arch Pathol Lab Med 2010; 134(9): 1261-70.
  13. Zeng C, Yu X, Lai J, Yang L, Chen S, Li Y. Overexpression of the long non-coding RNA PVT1 is correlated with leukemic cell proliferation in acute promyelocytic leukemia. J Hematol Oncol 2015; 8: 126.
  14. Cui M, You L, Ren X, Zhao W, Liao Q, Zhao Y. Long non-coding RNA PVT1 and cancer. Biochem Biophys Res Commun 2016; 471(1): 10-4.
  15. Colombo T, Farina L, Macino G, Paci P. PVT1: A rising star among oncogenic long noncoding RNAs. Biomed Res Int 2015; 2015: 1-10.
  16. Liu T, Yu T, Hu H, He K. Knockdown of the long non-coding RNA HOTTIP inhibits colorectal cancer cell proliferation and migration and induces apoptosis by targeting SGK1. Biomed Pharmacother 2018; 98: 286-96.
  17. Ren K, Xu R, Huang J, Zhao J, Shi W. Knockdown of long non-coding RNA KCNQ1OT1 depressed chemoresistance to paclitaxel in lung adenocarcinoma. Cancer Chemother Pharmacol 2017;
  18. (2): 243-50.
  19. Barata P, Sood AK, Hong DS. RNA-targeted therapeutics in cancer clinical trials: Current status and future directions. Cancer Treat Rev 2016; 50: 35-47.
  20. Lai J, Ozen A, Mouritzen P, Tolstrup N, Frandsen NM. Abstract PR14: Potent knock down of lncRNAs in vitro and in vivo with antisense LNA GapmeRs. Cancer Res 2016; 76(6 Suppl): R14.
  21. Alvarez-Dominguez JR, Hu W, Gromatzky AA, Lodish HF. Long noncoding RNAs during normal and malignant hematopoiesis. Int J Hematol 2014; 99(5): 531-41.
  22. Rodriguez-Malave NI, Rao DS. Long noncoding RNAs in hematopoietic malignancies. Brief Funct Genomics 2016; 15(3): 227-38.
  23. Guan Y, Kuo WL, Stilwell JL, Takano H, Lapuk AV, Fridlyand J, et al. Amplification of PVT1 contributes to the pathophysiology of ovarian and breast cancer. Clin Cancer Res 2007; 13(19): 5745-55.
  24. Takahashi Y, Sawada G, Kurashige J, Uchi R, Matsumura T, Ueo H, et al. Amplification of PVT-1 is involved in poor prognosis via apoptosis inhibition in colorectal cancers. Br J Cancer 2014; 110(1): 164-71.
  25. Iden M, Fye S, Li K, Chowdhury T, Ramchandran R, Rader JS. The lncRNA PVT1 contributes to the cervical cancer phenotype and associates with poor patient prognosis. PLoS One 2016; 11(5): e0156274.
  26. Wang F, Yuan JH, Wang SB, Yang F, Yuan SX, Ye C, et al. Oncofetal long noncoding RNA PVT1 promotes proliferation and stem cell-like property of hepatocellular carcinoma cells by stabilizing NOP2. Hepatology 2014; 60(4): 1278-90.
  27. Guo K, Yao J, Yu Q, Li Z, Huang H, Cheng J, et al. The expression pattern of long non-coding RNA PVT1 in tumor tissues and in extracellular vesicles of colorectal cancer correlates with cancer progression. Tumour Biol 2017; 39(4): 1010428317699122.
  28. Elmen J, Thonberg H, Ljungberg K, Frieden M, Westergaard M, Xu Y, et al. Locked nucleic acid (LNA) mediated improvements in siRNA stability and functionality. Nucleic Acids Res 2005; 33(1): 439-47.
  29. Parasramka MA, Maji S, Matsuda A, Yan IK, Patel T. Long non-coding RNAs as novel targets for therapy in hepatocellular carcinoma. Pharmacol Ther 2016; 161: 67-78.
Journal of Isfahan Medical School
Volume 36, Issue 477 - Serial Number 477
March and April 2018
Pages 439-445

Files

History

  • Receive Date: 17 April 2018
  • Revise Date: 23 May 2024
  • Accept Date: 06 April 2022

Share

How to cite

Statistics