Association between Altered Expression and Genetic Variations of Transforming Growth Factor β-Smad Pathway with Chronic Myeloid Leukemia
AbstractBackground: Chronic myeloid leukemia (CML) is a hematological disorder caused by fusion of BCR and ABL genes. BCR-ABL dependent and independent pathways play equally important role in CML. TGFβ-Smad pathway, an important BCR -ABL independent pathway, has scarce data in CML. Present study investigate the association between TGFβ-Smad pathway and CML.Materials and Methods: Sixty-four CML patients and age matched healthy controls (n=63) were enrolled in this study. Patients were segregated into responder and resistant groups depending on their response to Imatinib mesylate (IM). TGFβ1 serum levels were evaluated by ELISA and transcript levels of TGFβ1 receptors, SMAD4 and SMAD7 were evaluated by Real-Time PCR. Sequencing of exons and exon-intron boundaries of study genes was performed using Next Generation Sequencing (NGS) in 20 CML patients. Statistical analysis was performed using SPSS version 16.0.Results: TGFβ1 serum levels were significantly elevated (p = 0.02) and TGFβR2 and SMAD4 were significantly down-regulated (p = 0.012 and p = 0.043 respectively) in the patients. c.69A>G in TGFβ1, c.1024+24G>A in TGFβR1 and g.46474746C>T in SMAD7 were the most important genetic variants observed with their presence in 10/20, 8/20 and 7/20 patients respectively. In addition, TGFβR1 transcript levels were reduced in CML patients with c.69A>G mutation. None of the genes differed significantly in terms of expression or genetic variants between responder and resistant patient groups.Conclusion: Our findings demonstrate the role of differential expression and genetic variants of TGFβ-Smad pathway in CML. Decreased TGFβR2 and SMAD4 levels observed in the present study may be responsible for reduced tumor suppressive effects of this pathway in CML.
Nowell PC, Hungerford DA. A minute chromosome in human granulocytic leukemia. Science 1985; 142: 19.
Clarkson BD, Strife A, Wisniewski D, et al. New understanding of the pathogenesis of CML: a prototype of early neoplasia. Leukemia.1997; 11(9): 1404–28.
Druker BJ, Talpaz M, Resta DJ, et al.Efficacy and safety of a specific inhibitor of the BCR-ABL tyrosine in chronic myeloid leukemia. New Engl J Med. 2001; 344(14):1031-7.
Rice KN, Jamieson CH. Molecular pathways to CML stem cells. Int J Hematol. 2010; 91(5):748–52.
Dai Y, Rahmani M, Pei XY, et al. Bortezomib and flavopiridol interact synergistically to induce apoptosis in chronic myeloid leukemia cells resistant to Imatinibmesylate through both BCR/ABL-dependent and independent mechanisms. Blood. 2004; 104(2): 509-18.
Letterio JJ, Roberts AB. Regulation of immune responses by TGF-beta. Annu Rev Immunol 1998; 16: 137-61.
Massagué J, Blain SW, Lo RS. TGF beta signaling in growth control, cancer, and heritable disorders. Cell 2000; 103(2): 295–309.
Derynck R, Zhang YE. Smad-dependent and Smad-independent pathways in TGF-beta family signalling. Nature. 2003; 425(6958): 577–84.
Heldin CH, Miyazono K, ten Dijke P. TGF-beta signalling from cell membrane to nucleus through SMAD proteins. Nature.1997; 390(6659): 465–71.
Zhang Y, Derynck R. Regulation of Smadsignalling by protein associations and signaling crosstalk. Trends Cell Biol. 1999; 9(7): 274–9.
Pasche B. Role of Transforming Growth Factor Beta in Cancer. J Cell Physiol. 2001; 186(2): 153-68.
Myeroff LL, Parsons R, Kim SJ, et al. A transforming growth factor β receptor type II gene mutation common in colon and gastric but rare in endometrial cancers with microsatellite instability. Cancer Res. 1995; 55(23): 5545-7.
Imai Y, Tsurutani N, Oda H, et al. Genetic instability and mutation of the TGF-beta-receptor II in ampullary carcinomas. Int J Cancer. 1998; 76(3): 407-11.
Izumoto S, Arita N, Ohnishi T, et al. Microsatellite instability and mutated type II transforming growth factor-beta receptor gene in gliomas. Cancer Lett. 1997; 112(2): 251-6.
Paterson IC, Matthews JB, Huntley S, et al. Decreased expression of TGF- β cell surface receptors during progression of human oral squamous cell carcinoma. J Pathol. 2001; 193(4): 458-67.
Kim YH, Lee HS, Lee HJ, et al. Prognostic significance of the expression of SMAD4 and SMAD7 in human gastric carcinoma. Ann Oncol. 2004; 15(4): 574-80.
Singh P, Shrinivasan R, Wig JD, et al. A study of SMAD4, Smad6 and SMAD7 in surgically resected samples of Pancreatic ductal adenocarcinoma and their correlation with clinicopathological parameters and patients. BMC Res Notes. 2011; 4: 560.
Go JH. Altered expression of Smad proteins in T or NK cells lymphomas. Cancer Res Treat. 2008; 40(4): 197-201.
Andreef M, Wang R, Davis RE, et al. Proteomic, Gene Expression, and Micro-RNA Analysis of Bone Marrow Mesenchymal Stromal Cells in Acute Myeloid Leukemia Identifies Pro-Inflammatory, Pro-Survival Signatures In Vitro and In Vivo. Blood. 2013; 122(21): 3685.
Hamad A, Sahli Z, Sabban ME, et al. Emerging therapeutic strategies for targeting chronic myeloid leukemia stem cells. Stem Cells Int. 2013; 2013: 724360.
Kim SJ, Letterio J. Transforming growth factor-beta signaling in normal and malignant hematopoiesis. Leukemia. 2003; 17(9): 1731-7.
DeCoteau JF, Knaus PI, Yankelev H, et al. Loss of functional cell surface transforming growth factor beta (TGF-beta) type 1 receptor correlates with insensitivity to TGF-beta in chronic lymphocytic leukemia. ProcNatlAcadSci USA.1997; 94(11): 5877–81.
Le Bousse-Kerdilès MC, Chevillard S, Charpentier A,
et al. Differential expression of transforming growth factor-beta, basic fibroblast growth factor, and their receptors in CD34+ hematopoietic progenitor cells from patients with myelofibrosis and myeloid metaplasia. Blood.1996; 88(12): 4534–46.
Møller GM, Frost V, Melo JV, et al. Upregulation of the TGFbeta signaling pathway by Bcr-Abl: implications for haemopoietic cell growth and chronic myeloid leukaemia. FEBS Lett. 2007; 581(7): 1329-34.
Zhu X, Wang L, Zhang B, et al. TGF-beta1-induced PI3K/Akt/NF-kappaB/MMP9 signalling pathway is activated in Philadelphia chromosome-positive chronic myeloid leukaemiahemangioblasts.J Biochem. 2011; 149(4): 405-14.
K Naka, T Hoshii, T Muraguchi, et al. TGF-Β-FOXO signaling maintains leukaemia-initiating cells in chronic myeloid leukaemia.Nature. 2010; 463(7281): 676–80.
Liu X, Shan Y, Xue B. Int7G24A polymorphism (rs334354) and cancer risk. Arch Med Sci. 2013; 9(1): 3-7.
Ciftci R, Tas F, Yasasever CT, et al. High serum transforming growth factor beta 1 (TGFB1) level predicts better survival in breast cancer. Tumor Biol. 2014; 35(7): 6941-8.
Choi YJ, Kim N, Shin A, et al. Influence of TGFB1 C-509T polymorphism on gastric cancer risk associated with TGF-β1 expression in the gastric mucosa. Gastric Cancer.2015; 18(3): 526-37.
Wong TYH, Poon P, Chow KM, et al. Association of transforming growth factor-beta (TGF-beta) T869C (Leu 10 Pro) gene polymorphisms with type 2 diabetic nephropathy in Chinese. Kidney Intl. 2003; 63(5): 1831–35.
Pooja S, Francis A, Rajender S, et al. Strong impact of TGF-β1 gene polymorphisms on Breast cancer risk in Indian women: A case controls and population based study. PloS One. 2013; 8(10): e75979.
Wood NA, Thomson SC, Smith RM, et al. Identification of human TGF-beta1 signal (leader) sequence polymorphisms by PCR-RFLP. J Immunol Methods. 2000; 234(1-2): 117–22.
Smith PG, Tanaka H, Chantry A. A novel co-operative mechanism linking TGF β and Lyn kinase activation to Imatinib resistance in chronic myeloid leukemia cells. Oncotarget. 2012; 3(5): 518-24.
Rooke HM, Vitas MR, Crosier PS, et al. The TGF-β type II receptor in chronic myeloid leukemia: analysis of microsatellite regions and gene expression. Leukemia. 1999; 13(4): 535-41.
Tzur G, Israel A, Levy A, et al. Comprehensive gene and microRNA expression profiling reveals a role for microRNAs in human liver development. PloS One. 2009; 4(10): e7511.
Wu W, Tong Y, Wei X, et al. Association between Int7G24A rs334354 polymorphism and cancer risk: a meta-analysis of case-control studies. SciRep. 2015; 5:11350.
Liu L, Nie J, Chen L, et al. The oncogenic role of microRNA-130a/301a/454 in human colorectal cancer via targeting Smad4 expression. PLoS One. 2013; 8(2): e55532.
Bornstein S, White R, Malkoski S, et al. Smad4 loss in mice causes spontaneous head and neck cancer with increased genomic instability and inflammation. J Clin Invest. 2009; 119(11): 3408-19.
Qiao W, Li AG, Owens P, et al. Hair follicle defects and squamous cell carcinoma formation in Smad4 conditional knockout mouse skin. Oncogene. 2006; 25(2): 207-17.
Yang L, Mao C, Teng Y, et al. Targeted disruption of Smad4 in mouse epidermis results in failure of hair follicle cycling and formation of skin tumors. Cancer Res. 2005; 65(19): 8671-8.
Li W, Qiao W, Chen L, et al. Squamous cell carcinoma and mammary abscess formation through squamous metaplasia in Smad4/Dpc4 conditional knockout mice. Development. 2003; 130(24): 6143-53.
Xu X, Kobayashi S, Qiao W, et al. Induction of intra hepatic cholangio cellular carcinoma by liver-specific disruption of Smad4 and Pten in mice. J Clin Invest. 2006; 116(7): 1843-52.
Gao Y, Yang G, Weng T, et al. Disruption of Smad4 in odontoblasts causes multiple keratocysticodontogenic tumors and tooth malformation in mice. Mol Cell Biol. 2009; 29(21): 5941-51.
Zhang Y, Cao X, Jiang M, et al. Expression of Smad4 in leukemia cells. Zhongguo Shi Yan Xue Ye XueZaZhi. 2006; 14(4): 673-6.