Mencej Simona, Albagha Omar M E, Prezelj Janez, Kocjan Tomaz, Marc Janja
Faculty of Pharmacy, Chair of Clinical Biochemistry, University of Ljubljana, Askerceva cesta 7, SI-1000 Ljubljana, Slovenia.
J Mol Endocrinol. 2008 Jun;40(6):273-9. doi: 10.1677/JME-08-0003.
Tumour necrosis factor superfamily member 11 (TNFSF11) gene, that codes for receptor activator of nuclear factor-kappaB ligand, is one of the candidate genes for the genetic susceptibility to osteoporosis. As variations in the TNFSF11 gene promoter could alter its expression, the aim of the study was to evaluate the functional influence of three polymorphisms in the promoter and to investigate their association with bone mineral density (BMD) and biochemical markers in postmenopausal women. A total of 404 postmenopausal women were genotyped for the presence of TNFSF11 gene promoter polymorphisms -290C>T, -643C>T and -693G>C. Two common haplotypes, CCG and TTC, which occur in 44.3 and 49.3% of subjects respectively, were subjected to functional analysis. Amplified fragments were cloned into pGL3-basic reporter plasmid, which was co-transfected with pRL-TK plasmid into HEK293 cells. Dual luciferase reporter assay was performed. BMD and biochemical markers were measured. Reporter gene analysis showed significantly higher luciferase activity in CCG than in TTC haplotype (P=0.018). Both showed association with lumbar spine BMD (BMD-ls; P=0.005 and 0.007 for TTC and CCG respectively), whereas in femoral neck there was no association with BMD. In postmenopausal osteoporosis, association with BMD-ls was established in -290C>T, -643C>T and -693G>C (P values: 0.001, 0.041 and 0.013 respectively). Association with femoral neck BMD was shown in -693G>C (P=0.049). No association was found with biochemical markers in any of the groups. Our results suggest that in postmenopausal osteoporosis, TNFSF11 gene promoter polymorphisms -290C>T, -643C>T and -693G>C play a functional role in the genetic regulation of BMD.
肿瘤坏死因子超家族成员11（TNFSF11）基因编码核因子κB受体激活剂配体，是骨质疏松症遗传易感性的候选基因之一。由于TNFSF11基因启动子的变异可能会改变其表达，本研究的目的是评估启动子中三个多态性的功能影响，并研究它们与绝经后女性骨密度（BMD）和生化标志物的关联。对总共404名绝经后女性进行了TNFSF11基因启动子多态性-290C>T、-643C>T和-693G>C的基因分型。两种常见单倍型CCG和TTC，分别出现在44.3%和49.3%的受试者中，进行了功能分析。将扩增片段克隆到pGL3-基本报告质粒中，该质粒与pRL-TK质粒共转染到HEK293细胞中。进行双荧光素酶报告基因检测。测量BMD和生化标志物。报告基因分析显示CCG单倍型的荧光素酶活性显著高于TTC单倍型（P=0.018）。两者均与腰椎骨密度相关（TTC和CCG分别为P=0.005和0.007），而在股骨颈与骨密度无关联。在绝经后骨质疏松症中，-290C>T、-643C>T和-693G>C与腰椎骨密度相关（P值分别为0.001、0.041和0.013）。-693G>C与股骨颈骨密度相关（P=0.049）。在任何组中均未发现与生化标志物相关。我们的结果表明，在绝经后骨质疏松症中，TNFSF11基因启动子多态性-290C>T、-643C>T和-693G>C在骨密度的遗传调控中发挥功能作用。
