Hereditas (Beijing) (遗传) 2009/31:4 PP.374-380
To explore the mechanism of TBX5 abnormal expression in simple congenital heart disease (CHD), 100 CHD venous blood, 50 CHD heart tissues, and 5 non-CHD heart tissues were involved in this study. The mutation and methylation in the 1 200 bp region upstream of TBX5 gene were detected by high-performance liquid chromatog-raphy (DHPLC) and methylation-sensitive restriction endonuclease (MS-RE), respectively. The binding site of NKX2-5 to Tbx5 predicted by P-MATCH software was validated by EMSA (Electrophoretic mobility shift assay).Tbx5 gene expression in mouse cardiac muscle cell H9C2(2-1) transfected with NKX2-5 expression vector was evaluated. No mutation was found in all patients. Both non-CHD and CHD heart tissues had the same methylation in the two CpG islands. Exogenous Nkx2-5 efficiently activated the transcription of the endogenous Tbx5 gene in H9C2 (2-1) cells. EMSA showed that the special binding band appeared when Nkx2-5 existed. These results indicates that the down expres-sion of TBX5 might not be caused by mutation and methylation in the 1 200 bp region upstream of gene, and might be regulated by abnormal expression of NKX2-5 gene in heart muscle of CHD.
 Schott JJ, Benson DW, Basson CT, Pease W, Silberbach GM, Moak JP, Maron BJ, Seidman CE, Seidman JG. Con-genital heart disease caused by mutations in the transcrip-tion factor Nkx2-5.Science,1998, 281(5373): 108-111.
 Basson CT, Cowley GS, Solomon SD, Weissman B, Poznanski AK, Traill TA, Seidman JG, Seidman CE. The clinical and genetic spectrum of Holt-Oram syndrome.New Engl J Med, 1994, 330(13): 885-891.
 Dai YS, Cserjesi P, Markham BE, Molkentin JD. The transcription factors GATA4 and dHAND physically in-teract to synergistically activate cardiac gene expression through a p300-dependent mechanism.J Biol Chem, 2002, 277(27): 24390-24398.
 Garg V, Kathiriya IS, Barnes R, Schluterman MK, King IN, Butler CA, Rothrock CR, Eapen RS, Hirayama- Ya-mada K, Joo K, Matsuoka R, Cohen JC, Srivastava D. GATA4 mutations cause human congenital heart defects and reveal an interaction with TBX5.Nature, 2003, 424(6947): 443-447.
 Franco D, Campione M. The role of Pitx2 during cardiac development. Linking left-right signaling and congenital heart diseases.Trends Cardiovasc Med, 2003, 13(4): 157-163.
 Negro A, Brar BK, Lee KF. Essential roles of Her2/erbB2 in cardiac development and function.Recent Prog Horm Res, 2004, 59(1): 1-12.
 Lin Q, Schwarz J, Bucana C, Olson EN. Control of mouse cardiac morphogenesis and myogenesis by transcription factor MEF2C.Science, 1997, 276(5317): 1404-1407.
 Firulli AB, McFadden DG, Lin Q, Srivastava D, Olson EN. Heart and extra-embryonic mesodermal defects in mouse embryos lacking the bHLH transcription factor Hand1.Nat Genet, 1998, 18(3): 266-270.
 Riley P, Anson-Cartwright L, Cross JC. The Hand1 Bhlh transcription factor is essential for plancentation and car-diac morphogenesis.Nat Genet, 1998, 18(3): 271-275.
 Overbeek PA. Right and left go dHAND and eHAND.Nat Genet, 1997, 16(2): 119-121.
 Luft FC. Endothelin and cardiac development.J Mol Med, 2002, 80(11): 685-686.
 Bao ZZ, Bruneau BG, Seidman JG, Seidman CE, Cepko CL. Regulation of chamber-specific gene expression in the development heart by Irx4.Science, 1999, 283(4505): 1161-1164.
 Bruneau BG, Bao ZZ, Tanaka M, Schott JJ, Izumo S, Cepko CL, Seidman JG, Seidman CE. Cardiac expression of the ventricle-specific homeobox gene Irx4 is modulated by Nkx2-5 and dHAND.Dev Biol, 2000, 127(2): 266-277.
 Yamagishi H, Garg V, Matsuoka R, Thomas T, Srivastava D. A molecular pathway revealing a genetic basis for hu-man cardiac and craniofacial defects.Science, 1999, 283 (5405): 1158-1161.
 Bushdid PB, Osinska H, Waclaw RR, Molkentin JD, Yutzey KE. NFATc3 and NFATc4 are required for cardiac development and mitochondrial function.Circ Res, 2003, 92(12): 1305-1313.
 Lange AW, Molkentin JD, Yutzey KE. DSCR1 gene ex-pression is dependent on NFATc1 during cardiac valve formation and colocalizes with anomalous organ devel-opment in trisomy 16 mice.Dev Biol, 2004, 266(2): 346360.
 Cohen-Barak O, Yi Z, Hagiwara N, Monzen K, Komuro I, Brilliant MH. Sox6 regulation of cardiac myocyte devel-opment.Nucleic Acids Res, 2003, 31(20): 5941-5948.
 孙淼, 邱广蓉, 孙桂凤, 袁翼华, 孙开来. 人类单纯性先天性心脏病患者Nkx2-5基因突变及表达的研究. 中国医科大学学报, 2001, 30(5): 321-324.
 Liberatore CM, Searcy-Schrick R, Yutzey KE. Ventricular expression of tbx5 inhibits normal heart chamber devel-opment.Dev Biol, 2000, 223(1): 169-180.
 Huang T, Lock JE, Marshall AC, Basson C, Seidman JG, Seidman CE. Causes of clinical diversity in human TBX5 mutations.Quantitative Biol, 2002, 67: 115-120.
 宫立国, 邱广蓉, 邱广斌, 贺光, 孙开来. 人类单纯性先天性心脏病中TBX5基因的突变及表达研究. 遗传, 2003, 25(5): 39-43.
 Mori AD, Zhu Y, Vahora I, Nieman B, Koshiba-Takeuchi K, Davidson L, Pizard A, Seidman JG, Seidman CE, Chen XJ, Henkelman RM, Bruneau BG. Tbx5-dependent rheo-static control of cardiac gene expression and morphogene-sis.Dev Biol, 2006, 297(2): 566-586.
 Bruneau BG, Nemer G, Schmitt JP, Charron F, Robitaille L, Caron S, Conner DA, Gessler M, Nemer M, Seidman CE, Seidman JG. A murine model of Holt–Oram syndrome defines roles of the T-box transcription factor Tbx5 in cardiogenesis and disease.Cell, 2001, 106(6): 709-721.
 Lincoln J, Alfieri CM, Yutzey KE. Development of heart valve leaflets and supporting apparatus in chicken and mouse embryos.Dev Dyn, 2004, 230(2): 239-250.