The Effect of miR-372 on Genome Instability in MKN-45 Cell Line

Document Type : Original Article (s)

Authors

1 PhD Student, Department of Medical Genetics, School of Medical Sciences, Tarbiat Modares University, Tehran, Iran

2 Professor, Department of Medical Genetics, School of Medical Sciences, Tarbiat Modares University, Tehran, Iran

3 Associate Professor, Department of Hematology, School of Medical Sciences, Tarbiat Modares University, Tehran, Iran

Abstract

Background: Gastric cancer is one of the most common cancers in the world and the second leading cause of cancer mortality in humans. MicroRNAs are a group of endogenous RNA, small non-coding nucleotides in length of 21-23. Overexpression of miR-372 acts as an oncomir in various types of cancer via down-regulation of its target, LATS2. Down-regulation of LATS2 leads to the loss of cell cycle regulation, apoptosis inhibition, and increased proliferation rate of the cells.Methods: In this study, we increased the expression of miR-372 with lentivirus transduction inside the GC cell line MKN-45. After selection of positive cells, miR-372 and LATS2 expression levels were measured through real-time polymerase chain reaction (RT-PCR) assay. Cytochalasin B blocked (MN) assay was done to verify the presence or absence of MN for comparing genomic instability in treated cells compared to the controls.Findings: In the treated cells, compared with the controls, the amount of miR-372 expression significantly increased. Fold changes in 7, 14 and 21 days after the transduction were 7.85, 50.22 and 114.68, respectively (P = 0.030). In contrast to the control cells, the fold changes of LATS2 expression in these days were 0.39, 0.29 and 0.15, respectively (P = 0. 016). In addition, compared with control cells, the genomic instability of treated cells increased significantly (P < 0.001).Conclusion: These results indicate that in MKN-45 cell line, LATS2 is a target of miR-372. LATS2 is down-regulated with increased expression of miR-372. Reduce LATS2, leads to genomic instability during cell division and creates micronuclei and hence may be an important tumor suppressor.

Keywords

References

  1. Jemal A, Siegel R, Ward E, Hao Y, Xu J, Murray T, et al. Cancer statistics, 2008. CA Cancer J Clin 2008; 58(2): 71-96.
  2. Negrini M, Nicoloso MS, Calin GA. MicroRNAs and cancer--new paradigms in molecular oncology. Curr Opin Cell Biol 2009; 21(3): 470-9.
  3. Bartel DP. MicroRNAs: genomics, biogenesis, mechanism, and function. Cell 2004; 116(2): 281-97.
  4. Rana TM. Illuminating the silence: understanding the structure and function of small RNAs. Nat Rev Mol Cell Biol 2007; 8(1): 23-36.
  5. Shukla GC, Singh J, Barik S. MicroRNAs: Processing, Maturation, Target Recognition and Regulatory Functions. Mol Cell Pharmacol 2011; 3(3): 83-92.
  6. Felekkis K, Touvana E, Stefanou C, Deltas C. microRNAs: a newly described class of encoded molecules that play a role in health and disease. Hippokratia 2010; 14(4): 236-40.
  7. Takuno S, Innan H. Evolution of complexity in miRNA-mediated gene regulation systems. Trends Genet 2008; 24(2): 56-9.
  8. Di LG, Croce CM. Roles of small RNAs in tumor formation. Trends Mol Med 2010; 16(6): 257-67.
  9. Farazi TA, Hoell JI, Morozov P, Tuschl T. MicroRNAs in human cancer. Adv Exp Med Biol 2013; 774: 1-20.
  10. Wang D, Qiu C, Zhang H, Wang J, Cui Q, Yin Y. Human microRNA oncogenes and tumor suppressors show significantly different biological patterns: from functions to targets. PLoS One 2010; 5(9).
  11. Calin GA, Croce CM. MicroRNA signatures in human cancers. Nat Rev Cancer 2006; 6(11): 857-66.
  12. Katada T, Ishiguro H, Kuwabara Y, Kimura M, Mitui A, Mori Y, et al. microRNA expression profile in undifferentiated gastric cancer. Int J Oncol 2009; 34(2): 537-42.
  13. Guo J, Miao Y, Xiao B, Huan R, Jiang Z, Meng D, et al. Differential expression of microRNA species in human gastric cancer versus non-tumorous tissues. J Gastroenterol Hepatol 2009; 24(4): 652-7.
  14. Song MY, Pan KF, Su HJ, Zhang L, Ma JL, Li JY, et al. Identification of serum microRNAs as novel non-invasive biomarkers for early detection of gastric cancer. PLoS One 2012; 7(3): e33608.
  15. Yin Y, Li J, Chen S, Zhou T, Si J. MicroRNAs as diagnostic biomarkers in gastric cancer. Int J Mol Sci 2012; 13(10): 12544-55.
  16. Voorhoeve PM, le SC, Schrier M, Gillis AJ, Stoop H, Nagel R, et al. A genetic screen implicates miRNA-372 and miRNA-373 as oncogenes in testicular germ cell tumors. Cell 2006; 124(6): 1169-81.
  17. Cho WJ, Shin JM, Kim JS, Lee MR, Hong KS, Lee JH, et al. miR-372 regulates cell cycle and apoptosis of ags human gastric cancer cell line through direct regulation of LATS2. Mol Cells 2009; 28(6): 521-7.
  18. Yamashita S, Yamamoto H, Mimori K, Nishida N, Takahashi H, Haraguchi N, et al. MicroRNA-372 is associated with poor prognosis in colorectal cancer. Oncology 2012; 82(4): 205-12.
  19. Yu SL, Chen HY, Chang GC, Chen CY, Chen HW, Singh S, et al. MicroRNA signature predicts survival and relapse in lung cancer. Cancer Cell 2008; 13(1): 48-57.
  20. Stelzer Y, Sagi I, Benvenisty N. Involvement of parental imprinting in the antisense regulation of onco-miR-372-373. Nat Commun 2013; 4: 2724.
  21. Visser S, Yang X. LATS tumor suppressor: a new governor of cellular homeostasis. Cell Cycle 2010; 9(19): 3892-903.
  22. (22) Aylon Y, Michael D, Shmueli A, Yabuta N, Nojima H, Oren M. A positive feedback loop between the p53 and Lats2 tumor suppressors prevents tetraploidization. Genes Dev 2006; 20(19): 2687-700.
  23. Yabuta N, Okada N, Ito A, Hosomi T, Nishihara S, Sasayama Y, et al. Lats2 is an essential mitotic regulator required for the coordination of cell division. J Biol Chem 2007; 282(26): 19259-71.
  24. Kim YK, Yu J, Han TS, Park SY, Namkoong B, Kim DH, et al. Functional links between clustered microRNAs: suppression of cell-cycle inhibitors by microRNA clusters in gastric cancer. Nucleic Acids Res 2009; 37(5): 1672-81.
  25. Belair C, Baud J, Chabas S, Sharma CM, Vogel J, Staedel C, et al. Helicobacter pylori interferes with an embryonic stem cell micro RNA cluster to block cell cycle progression. Silence 2011; 2(1): 7.
  26. Wang Z, Yang Ch. Circulating microRNAs as diagnostic and prognostic biomarkers for gastric cancer-opportunities and challenges. Am J Digest Dis 2014;1(1): 44-9.
  27. Medina PP, Slack FJ. microRNAs and cancer: an overview. Cell Cycle 2008; 7(16): 2485-92.
  28. Liao Y L, Tsai K W, Wen-chang Lin. miRNAs in gastric cancer. In: Lotfy M, editor. Gastric Carcinoma - Molecular Aspects and Current Advances. Rijeka, Croatia: Intech; 2011. p. 85-104.
  29. Bartels CL, Tsongalis GJ. MicroRNAs: novel biomarkers for human cancer. Clin Chem 2009; 55(4): 623-31.
  30. Li G, Zhang Z, Tu Y, Jin T, Liang H, Cui G, et al. Correlation of microRNA-372 upregulation with poor prognosis in human glioma. Diagn Pathol 2013; 8: 1.
  31. Cairo S, Wang Y, de Reynies A, Duroure K, Dahan J, Redon MJ, et al. Stem cell-like micro-RNA signature driven by Myc in aggressive liver cancer. Proc Natl Acad Sci U S A 2010; 107(47): 20471-6.
  32. Tian RQ, Wang XH, Hou LJ, Jia WH, Yang Q, Li YX, et al. MicroRNA-372 is down-regulated and targets cyclin-dependent kinase 2 (CDK2) and cyclin A1 in human cervical cancer, which may contribute to tumorigenesis. J Biol Chem 2011; 286(29): 25556-63.
  33. Takaishi S, Okumura T, Tu S, Wang SS, Shibata W, Vigneshwaran R, et al. Identification of gastric cancer stem cells using the cell surface marker CD44. Stem Cells 2009; 27(5): 1006-20.
  34. Golestaneh AF, Atashi A, Langroudi L, Shafiee A, Ghaemi N, Soleimani M. miRNAs expressed differently in cancer stem cells and cancer cells of human gastric cancer cell line MKN-45. Cell Biochem Funct 2012; 30(5): 411-8.
  35. McPherson JP, Tamblyn L, Elia A, Migon E, Shehabeldin A, Matysiak-Zablocki E, et al. Lats2/Kpm is required for embryonic development, proliferation control and genomic integrity. EMBO J 2004; 23(18): 3677-88.
  36. Duensing S. A tentative classification of centrosome abnormalities in cancer. Cell Biol Int 2005; 29(5): 352-9.
  37. Salisbury JL, Whitehead CM, Lingle WL, Barrett SL. Centrosomes and cancer. Biol Cell 1999; 91(6): 451-60.
Journal of Isfahan Medical School
Volume 32, Issue 311 - Serial Number 311
September and October 2015
Pages 2035-2047

Files

History

  • Receive Date: 10 December 2014
  • Revise Date:
  • Accept Date: 06 April 2022

Share

How to cite

Statistics