Advertisement

Study of microRNA Profile as a Molecular Biomarker in Egyptian Chronic Lymphocytic Leukemia

  • Nahla Mohamed Gamal Farahat
  • Dalal Mohamed Nasr El Din Elkaffash
  • Ashraf Hussein Alghandour
  • Rania Shafik SwelemEmail author
  • Reham Abdel Haleem Abo El-Wafa
Original Article
  • 15 Downloads

Abstract

MicroRNAs target mRNAs for cleavage or translational repression. They play a critical role in the progression of malignancies and leukemias including chronic lymphocytic leukemia (CLL). However, microRNA expression levels in Egyptian patients with CLL, and their prognostic value remain elusive. Our main aim was to assess the expression pattern of a panel of microRNAs in CLL patients to create an informative microRNA profile. The study subjects were 40 newly diagnosed CLL patients of both sexes and 40 age and sex matched controls. The expression levels of 12 microRNAs were evaluated by qRT-PCR, including miR-15a, 16, 23b, 24, 29a, 29c, 34a, 146a, 155, 181a, 195, and 221. Flow cytometry was used to determine the expression levels of BCL2, CD38, and ZAP-70 in CLL patients. We identified various degrees of upregulated miRNAs (miR-29a, miR-29c, miR-34a, miR-155, miR-146a, and miR-195) and down-regulated ones (miR-15a, miR-16, miR-23b, miR-24, miR-181a, and miR-221) in CLL patients relative to controls. The mean fluorescence intensity ratio (MFI-R) of BCL2 was recorded and was significantly upregulated in CLL patients compared with normal controls. In addition, inverse correlations were observed between microRNAs (miR-15a, miR-16, miR-155, and miR-195) and BCL2 MFI-R while positive correlations were observed between miR-29a and miR-29c, and BCL2 MFI-R. These findings suggest that these miRNAs regulate BCL2 levels. Moreover, we found that miR-15a, miR-16, miR-155, miR-181a, miR-195 and miR-221 were significantly upregulated, while miR-29a and miR-29c were significantly downregulated in ZAP-70 positive CLL patients. Various miRNAs may play an important role in the pathogenesis of CLL and have the potential to be used for the prognosis of patients with CLL.

Keywords

miRNA CLL BCL-2 ZAP-70 CD38 

Notes

Acknowledgements

The authors would like to thank all participating technicians, and our patients, who without them this work could not have been accomplished.

Compliance with Ethical Standards

Conflict of interest

The authors declare that they have no conflict of interests.

Ethical Approval

All procedures performed in our study were in accordance with the ethical standards of our institution and national and with the 1975 Helsinki declaration as revised in 2008.

Supplementary material

12288_2018_1000_MOESM1_ESM.tif (53 kb)
Supplementary Fig. 1 Comparison between cases and control groups regarding miRNAs expression levels (TIFF 52 kb)
12288_2018_1000_MOESM2_ESM.tif (25 kb)
Supplementary Fig. 2 Relation between Lymphadenopathy in CLL patients and A) miR-23b expression, B) miR-221 expression (TIFF 25 kb)
12288_2018_1000_MOESM3_ESM.tif (99 kb)
Supplementary Fig. 3 Relation between Splenomegaly and miRNAs expression in CLL patients (TIFF 98 kb)
12288_2018_1000_MOESM4_ESM.tif (134 kb)
Supplementary Fig. 4 Box plot showing Relation between modified Rai stages and miRNAs expression levels in CLL patients (TIFF 134 kb)
12288_2018_1000_MOESM5_ESM.tif (133 kb)
Supplementary Fig. 5 Correlations between miRNAs expression levels and BCL2 MFI-R (TIFF 132 kb)
12288_2018_1000_MOESM6_ESM.tif (55 kb)
Supplementary Fig. 6 Relation between LDT and miRNAs expression levels in CLL patients (TIFF 54 kb)
12288_2018_1000_MOESM7_ESM.tif (53 kb)
Supplementary Fig. 7 Relation between treatment requirements and miRNAs expression levels in CLL patients (TIFF 53 kb)
12288_2018_1000_MOESM8_ESM.doc (42 kb)
Supplementary material 8 (DOC 42 kb)
12288_2018_1000_MOESM9_ESM.doc (34 kb)
Supplementary material 9 (DOC 34 kb)

References

  1. 1.
    Cheson BD, Bennett JM, Grever M et al (1996) National Cancer Institute-sponsored Working Group guidelines for chronic lymphocytic leukemia: revised guidelines for diagnosis and treatment. Blood 87:4990–4997Google Scholar
  2. 2.
    Chiorazzi N, Rai KR, Ferrarini M (2005) Chronic lymphocytic leukemia. N Engl J Med 352:804–815CrossRefGoogle Scholar
  3. 3.
    Novina CD, Sharp PA (2004) The RNAi revolution. Nature 430:161–164CrossRefGoogle Scholar
  4. 4.
    Lu J, Getz G, Miska EA et al (2005) MicroRNA expression profiles classify human cancers. Nature 435:834–838CrossRefGoogle Scholar
  5. 5.
    Chen CZ, Lodish HF (2005) MicroRNAs as regulators of mammalian hematopoiesis. Semin Immunol 17:155–165CrossRefGoogle Scholar
  6. 6.
    Papakonstantinou N, Ntoufa S, Chartomatsidou E et al (2013) Differential microRNA profiles and their functional implications in different immunogenetic subsets of chronic lymphocytic leukemia. Mol Med 19:115–123CrossRefGoogle Scholar
  7. 7.
    Yang M, Mattes J (2008) Discovery, biology and therapeutic potential of RNA interference, microRNA and antagomirs. Pharmacol Ther 117(1):94–104CrossRefGoogle Scholar
  8. 8.
    Müller-Hermelink H, Montserrat E, Catovsky D et al (2008) Chronic lymphocytic leukaemia/small lymphocytic lymphoma. In: Jaffe ES, Harris NL, Stein H, Vardiman JW (eds) World Health Organization classification of tumours: pathology and genetics of tumours of haematopoietic and lymphoid tissues. IARC, Lyon, pp 180–182Google Scholar
  9. 9.
    Hallek M, Cheson BD, Catovsky D et al (2008) Guidelines for the diagnosis and treatment of chronic lymphocytic leukemia: a report from the International Workshop on chronic lymphocytic leukemia updating the National Cancer Institute-Working Group 1996 guidelines. Blood 111:5446–5456CrossRefGoogle Scholar
  10. 10.
    Ward BP, Tsongalis GJ, Kaur P (2011) MicroRNAs in chronic lymphocytic leukemia. Exp Mol Pathol 90(2):173–178CrossRefGoogle Scholar
  11. 11.
    Nicoloso MS, Kipps TJ, Croce CM et al (2007) MicroRNAs in the pathogeny of chronic lymphocytic leukaemia. Br J Haematol 139(5):709–716CrossRefGoogle Scholar
  12. 12.
    Nana-Sinkam SP, Croce CM (2010) MicroRNA in chronic lymphocytic leukemia: transitioning from laboratory-based investigation to clinical application. Cancer Genet Cytogenet 203(2):127–133CrossRefGoogle Scholar
  13. 13.
    Calin GA, Liu CG, Sevignani C et al (2004) MicroRNA profiling reveals distinct signatures in B cell chronic lymphocytic leukemias. Proc Natl Acad Sci USA 101:11755–11760CrossRefGoogle Scholar
  14. 14.
    Zhu DX, Zhu W, Fang C et al (2012) miR-181a/b significantly enhances drug sensitivity in chronic lymphocytic leukemia cells via targeting multiple anti-apoptosis genes. Carcinogenesis 33(7):1294–1301CrossRefGoogle Scholar
  15. 15.
    Zanette DL, Rivadavia F, Molfetta GA et al (2007) miRNA expression profiles in chronic lymphocytic and acute lymphocytic leukemia. Braz J Med Biol Res 40(11):1435–1440CrossRefGoogle Scholar
  16. 16.
    Pallasch CP (2009) MiRNA deregulation by epigenetic silencing disrupts suppression of the oncogene PLAG1 in chronic lymphocytic leukemia. Blood 114:3255–3264CrossRefGoogle Scholar
  17. 17.
    McCarthy BA, Boyle E, Wang XP et al (2008) Surface expression of Bcl-2 in chronic lymphocytic leukemia and other B-cell leukemias and lymphomas without a breakpoint t(14;18). Mol Med 14(9–10):618–627Google Scholar
  18. 18.
    Pepper C, Hoy T, Bentley P (1998) Elevated Bcl-2/Bax are a consistent feature of apoptosis resistance in B-cell chronic lymphocytic leukaemia and are correlated with in vivo chemoresistance. Leuk Lymphoma 28(3–4):355–361CrossRefGoogle Scholar
  19. 19.
    Cimmino A, Calin GA, Fabbri M et al (2005) miR-15 and miR-16 induce apoptosis by targeting BCL2. Proc Natl Acad Sci USA 102:13944–13949CrossRefGoogle Scholar
  20. 20.
    Zhu D-X (2010) Aberrant microRNA expression in Chinese patients with chronic lymphocytic leukemia. Leuk Res 4076:1–5Google Scholar
  21. 21.
    Neilson JR, Zheng GX, Burge CB et al (2007) Dynamic regulation of miRNA expression in orderer stages of cellular development. Genes Dev 21:578–589CrossRefGoogle Scholar
  22. 22.
    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 353:1793–1801CrossRefGoogle Scholar
  23. 23.
    Fulci V, Chiaretti S, Goldoni M et al (2007) Quantitative technologies establish a novel microRNA profile of chronic lymphocytic leukemia. Blood 109(11):4944–4951CrossRefGoogle Scholar
  24. 24.
    Debernardi S, Skoulakis S, Molloy G et al (2007) MicroRNA miR-181a correlates with morphological sub-class of acute myeloid leukaemia and the expression of its target genes in global genome-wide analysis. Leukemia 21(5):912–916CrossRefGoogle Scholar
  25. 25.
    Pichiorri F, Suh SS, Ladetto M et al (2008) MicroRNAs regulate critical genes associated with multiple myeloma pathogenesis. Proc Natl Acad Sci USA 105(35):12885–12890CrossRefGoogle Scholar
  26. 26.
    Taylor MA, Sossey-Alaoui K, Thompson CL et al (2013) TGF-beta upregulates miR-181a expression to promote breast cancer metastasis. J Clin Invest 123(1):150–163CrossRefGoogle Scholar
  27. 27.
    Chen G, Zhu W, Shi D et al (2010) MicroRNA-181a sensitizes human malignant glioma U87MG cells to radiation by targeting Bcl-2. Oncol Rep 23(4):997–1003Google Scholar
  28. 28.
    He L, He X, Lowe SW et al (2007) microRNAs join the p53 network—another piece in the tumour-suppression puzzle. Nat Rev Cancer 7(11):819–822CrossRefGoogle Scholar
  29. 29.
    Neilsen PM, Noll JE, Mattiske S et al (2013) Mutant p53 drives invasion in breast tumors through up-regulation of miR-155. Oncogene 32(24):2992–3000CrossRefGoogle Scholar
  30. 30.
    Moussay E, Palissot V, Vallar L et al (2010) Determination of genes and microRNAs involved in the resistance to fludarabine in vivo in chronic lymphocytic leukemia. Mol Cancer 9:115CrossRefGoogle Scholar
  31. 31.
    Zenz T, Mohr J, Eldering E et al (2009) miR-34a as part of the resistance network in chronic lymphocytic leukemia. Blood 113(16):3801–3808CrossRefGoogle Scholar
  32. 32.
    Wang Y, Zhang X, Li H et al (2013) The role of miRNA-29 family in cancer. Eur J Cell Biol 92(3):123–128CrossRefGoogle Scholar
  33. 33.
    Stamatopoulos B, Meuleman N, Haibe-Kains B et al (2009) MicroRNA-29c and microRNA-223 down-regulation has in vivo significance in chronic lymphocytic leukemia and improves disease risk stratification. Blood 113:5237–5245CrossRefGoogle Scholar

Copyright information

© Indian Society of Hematology and Blood Transfusion 2018

Authors and Affiliations

  • Nahla Mohamed Gamal Farahat
    • 1
  • Dalal Mohamed Nasr El Din Elkaffash
    • 1
  • Ashraf Hussein Alghandour
    • 2
  • Rania Shafik Swelem
    • 1
    Email author
  • Reham Abdel Haleem Abo El-Wafa
    • 1
  1. 1.Clinical and Chemical Pathology Department, Faculty of MedicineAlexandria UniversityAzaritaEgypt
  2. 2.Internal Medicine (Hematology), Faculty of MedicineAlexandria UniversityAzaritaEgypt

Personalised recommendations