AKT family and miRNAs expression in IL-2-induced CD4+T cells

Document Type: Original Article


1 Department of Genetics, College of Science, Rasht Branch, Islamic Azad University, Rasht, Iran

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

3 Department of Molecular Medicine, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran

4 National Cell Bank of Iran, Pasteur Institute of Iran, Tehran, Iran

5 Department of Medical Genetics, Faculty of Medicine, Tehran University of Medical Sciences, Tehran, Iran


Objective(s): Study of non-coding RNAs is considerable to elucidate principal biological questions or design new therapeutic strategies. miRNAs are a group of non-coding RNAs that their functions in PI3K/AKT signaling and apoptosis pathways after T cell activation is not entirely clear. Herein, miRNAs expression and their putative targets in the mentioned pathways were studied in the activated CD4+T cells.
Materials and Methods: Herein, proliferation rate and IL-2 secretion were measured in treated and untreated cells by IL-2. Putative targets of up-regulated miRNAs were predicted by bioinformatics approaches in the apoptotic and PI3K/AKT signaling pathways. Then the expression of two putative targets was evaluated by quantitative RT-PCR. 
Results: Proliferation rate of treated cells by IL-2 increased in a dose- and time- dependent manner. Naive and activated CD4+T cells  induced by different dose of IL-2 secreted abundant amounts of IL-2. Also, in IL-2 un-induced cells (IL-2 depleted cells) after 3 days, decrease of proliferation has been shown. In silico analysis predicted putative targets of up-regulated miRNAs such as AKT1, AKT3 and apoptotic genes in the activated cells induced or un-induced by IL-2. Decrease of AKT3 was shown by Q-RT-PCR as a potential target of miRNAs overexpressed in IL-2 depleted cells. But there was no significant difference in AKT1 expression in two cell groups.
Conclusion:  Our analysis suggests that decrease of AKT3 was likely controlled via up-regulation of specific miRNAs in IL-2 depleted cells. Also it seems that miRNAs play role in induction of different apoptosis pathways in IL-2 induced and un-induced cells.


1.   Androulidaki A, Iliopoulos D, Arranz A, Doxaki C, Schworer S, Zacharioudaki V, et al. The kinase Akt1 controls macrophage response to lipopolysaccharide by regulating microRNAs. Immunity 2009; 31:220-231.

2. Juntilla MM, Wofford JA, Birnbaum MJ, Rathmell JC, Koretzky GA. Akt1 and Akt2 are required for alphabeta thymocyte survival and differentiation. Proc Natl Acad Sci USA 2007; 104:12105-12110.

3. Amaravadi R, Thompson CB. The survival kinases Akt and Pim as potential pharmacological targets. J Clin Invest 2005; 115:2618-2624.

4.   Lockyer HM, Tran E, Nelson BH. STAT5 is essential for Akt/p70S6 kinase activity during IL-2-induced lymphocyte proliferation. J Immunol 2007; 179:5301-5308.

5. Fung MM, Rohwer F, McGuire KL. IL-2 activation of a PI3K-dependent STAT3 serine phosphorylation pathway in primary human T cells. Cell Signall 2003; 15:625-636.

6.   Ahmed NN, Grimes HL, Bellacosa A, Chan TO, Tsichlis PN. Transduction of interleukin-2 antiapoptotic and proliferative signals via Akt protein kinase. Proc Natl Acad Sci U S A 1997; 94:3627-3632.

7. Bauer B, Baier G. Protein kinase C and AKT/protein kinase B in CD4+ T-lymphocytes: new partners in TCR/CD28 signal integration. Mol Immunol 2002; 38:1087-1099.

8.   Xiao C, Rajewsky K. MicroRNA control in the immune system: basic principles. Cell 2009; 136:26-36.

9.   Calin GA, Croce CM. MicroRNA signatures in human cancers. Nat Rev Cancer 2006; 6:857-866.

10. Lindsay MA. microRNAs and the immune response. Trends  Immunol 2008; 29:343-351.

11. Furer V, Greenberg JD, Attur M, Abramson SB, Pillinger MH. The role of microRNA in rheumatoid arthritis and other autoimmune diseases. Clin Immunol 2010; 136:1-15.

12. Wang Z, Li Y, Kong D, Ahmad A, Banerjee S, Sarkar FH. Cross-talk between miRNA and Notch signaling pathways in tumor development and progression. Cancer Lett 2010; 292:141-148.

13. Sonkoly E, Stahle M, Pivarcsi A. MicroRNAs and immunity: novel players in the regulation of normal immune function and inflammation. Semin Cancer Biol 2008; 18:131-140.

14. Molnar V, Tamasi V, Bakos B, Wiener Z, Falus A. Changes in miRNA expression in solid tumors: an miRNA profiling in melanomas. Semin Cancer Biol 2008; 18:111-122.

15. Gartel AL, Kandel ES. miRNAs: Little known mediators of oncogenesis. Semin Cancer Biol 2008; 18:103-110.

16. Babashah S, Sadeghizadeh M, Hajifathali A, Tavirani MR, Zomorod MS, Ghadiani M, et al. Targeting of the signal transducer Smo links microRNA-326 to the oncogenic Hedgehog pathway in CD34 CML stem/progenitor cells. Int J Cancer 2013; 133:579-589

17. Babashah S, Sadeghizadeh M, Tavirani MR, Farivar S, Soleimani M. Aberrant microRNA expression and its implications in the pathogenesis of leukemias. Cell Oncol (Dordr) 2012; 35:317-334.

18. Tili E, Michaille JJ, Calin GA. Expression and function of micro-RNAs in immune cells during normal or disease state. Int J Med Sci 2008; 5:73-79.

19. Almeida MI, Reis RM, Calin GA. MicroRNA history: discovery, recent applications, and next frontiers. Mutat Res 2011; 717:1-8.

20. Dai R, Ahmed SA. MicroRNA, a new paradigm for understanding immunoregulation, inflammation, and autoimmune diseases. Transl Res 2011; 157:163-179.

21. Du J, Yang H, Zhang D, Wang J, Guo H, Peng B, et al. Structural basis for the blockage of IL-2 signaling by therapeutic antibody basiliximab. J Immunol 2010; 184:1361-1368.

22. Shu L, Yin W, Zhuang H, Hua Z. Comparison of gene expression profiles in mouse primary T cells under normal and prolonged activation. Blood Cells Mol Dis 2006; 37:64-75.

23. Ranji N, Sadeghizadeh M, Shokrgozar MA, Bakhshandeh B, Karimipour M, Amanzadeh A, et al. MiR-17-92 cluster: an apoptosis inducer or proliferation enhancer. Mol Cell Biochem 2013; 380:229-238.

24. Jolly C, Sattentau QJ. Regulated secretion from CD4+ T cells. Trends Immunol 2007; 28:474-481.

25. Sakaguchi S, Wing K, Onishi Y, Prieto-Martin P, Yamaguchi T. Regulatory T cells: how do they suppress immune responses? Int Immunol 2009; 21:1105-1111.

26. Hieronymus T, Blank N, Gruenke M, Winkler S, Haas JP, Kalden JR, et al. CD 95-independent mechanisms of IL-2 deprivation-induced apoptosis in activated human lymphocytes. Cell Death Differ 2000; 7:538-547.

27. Tsitsiou E, Lindsay MA. microRNAs and the immune response. Curr Opin Pharmacol 2009; 9:514-520.

28. Baltimore D, Boldin MP, O'Connell RM, Rao DS, Taganov KD. MicroRNAs: new regulators of immune cell development and function. Nat Immunol 2008; 9:839-845.

29. Carissimi C, Fulci V, Macino G. MicroRNAs: novel regulators of immunity. Autoimmun Rev 2009; 8:520-524.

30. Madhunapantula SV, Desai D, Sharma A, Huh SJ, Amin S, Robertson GP. PBISe, a novel selenium-containing drug for the treatment of malignant melanoma. Mol Cancer Therapeut 2008; 7:1297-1308.

31. Inamdar GS, Madhunapantula SV, Robertson GP. Targeting the MAPK pathway in melanoma: why some approaches succeed and other fail. Biochem Pharm 2010; 80:624-637.

32. Matheny RW, Jr, Adamo ML. Role of Akt isoforms in IGF-I-mediated signaling and survival in myoblasts. Biochem Biophys Res Commun 2009; 389:117-121.

33. Liu X, Shi Y, Birnbaum MJ, Ye K, De Jong R, Oltersdorf T, et al. Quantitative analysis of anti-apoptotic function of Akt in Akt1 and Akt2 double knock-out mouse embryonic fibroblast cells under normal and stressed conditions.  J Biol Chem 2006; 281:31380-31388.

34. Reuter S, Eifes S, Dicato M, Aggarwal BB, Diederich M. Modulation of anti-apoptotic and survival pathways by curcumin as a strategy to induce apoptosis in cancer cells. Biochem Pharmacol 2008; 76:1340-1351.

35. Kelly E, Won A, Refaeli Y, Van Parijs L. IL-2 and related cytokines can promote T cell survival by activating AKT. J Immunol 2002; 168:597-603.

36. Lenardo MJ. Fas and the art of lymphocyte maintenance. J Exp Med 1996; 183:721-724.

37. Snow AL, Oliveira JB, Zheng L, Dale JK, Fleisher TA, Lenardo MJ. Critical role for BIM in T cell receptor restimulation-induced death. Biol Direct 2008; 3:34.

38. Dewson G, Kluck RM. Bcl-2 family-regulated apoptosis in health and disease. Cell Health Cytoskeleton 2010; 2:9-22.

39. Bosque A, Marzo I, Naval J, Anel A. Apoptosis by IL-2 deprivation in human CD8+ T cell blasts predominates over death receptor ligation, requires Bim expression and is associated with Mcl-1 loss. Mol Immunol 2007; 44:1446-1453.

40. Downward J. PI 3-kinase, Akt and cell survival. Semin Cell Dev Biol 2004; 15:177-182.