大骨节病差异表达基因PAPSS2对成骨细胞矿化及碱性磷酸酶活性的影响

被引:8
作者
王伟卓 [1 ]
程一钊 [1 ]
郭雄 [2 ]
王坤正 [1 ]
袁普卫 [3 ]
机构
[1] 西安交通大学医学院第二附属医院骨科
[2] 西安交通大学医学院环境与疾病相关基因教育部重点实验室及卫生部微量元素与地方病重点实验室
[3] 陕西中医学院附属医院骨科
关键词
大骨节病; 成骨细胞; 基因功能; 碱性磷酸酶; 骨关节炎; 逆转录病毒; PAPSS2; 细胞矿化;
D O I
暂无
中图分类号
R684.1 [大骨节病];
学科分类号
100220 [骨科学];
摘要
目的探讨大骨节病差异表达基因PAPSS2在MC3T3-E1细胞成骨样分化过程中的表达及其对碱性磷酸酶(ALP)活性和细胞矿化的影响。方法培养MC3T3-E1细胞并诱导成骨分化,观察PAPSS2在成骨细胞分化过程中的基因及蛋白表达情况;用慢病毒介导的RNA干扰(RNAi)技术或逆转录病毒转染高表达载体,分别构建PAPSS2低表达慢病毒及逆转录病毒高表达载体包装,滴度测定,验证细胞转染效率;分别转染MC3T3-E1细胞并设空白对照组,观察PAPSS2沉默或高表达后ALP活性的变化及细胞成骨矿化的影响。结果 PAPSS2在成骨细胞分化过程中mRNA及蛋白均随着矿化过程表达增加。用慢病毒介导的RNAi技术,成功降低了MC3T3-E1成骨细胞中PAPSS2的表达,使用逆转录病毒转染高表达载体显著增加PAPSS2基因的表达。PAPSS2沉默表达时,ALP活性和细胞矿化作用显著降低,高表达后其作用相反。结论 PAPSS2可调节成骨细胞ALP活性和细胞矿化作用,其异常表达可能与大骨节病骨骼病变相关。
引用
收藏
页码:175 / 181
页数:7
相关论文
共 10 条
[1]
Comparative analysis of gene expression profiles between the normal human cartilage and the one with endemic osteoarthritis.[J].W.Z. Wang;X. Guo;C. Duan;W.J. Ma;Y.G. Zhang;P. Xu;Z.Q. Gao;Z.F. Wang;H. Yan;Y.F. Zhang;Y.X. Yu;J.C. Chen;M.J. Lammi.Osteoarthritis and Cartilage.2008, 1
[2]
Human 3'‐phosphoadenosine 5'‐phosphosulfate (PAPS) Synthase: Biochemistry; Molecular Biology and Genetic Deficiency.[J].KVVenkatachalam.IUBMB Life.2008, 1
[3]
The effect of adsorbed vitamin D and K to hydroxyapatite on ALP activity of MC3T3-E1 cell [J].
Ozeki, K. ;
Aoki, H. ;
Fukui, Y. .
JOURNAL OF MATERIALS SCIENCE-MATERIALS IN MEDICINE, 2008, 19 (04) :1753-1757
[4]
Expression profile of Papss2 (3′-phosphoadenosine 5′-phosphosulfate synthase 2) during cartilage formation and skeletal development in the mouse embryo [J].
Stelzer, Christiane ;
Brimmer, Annette ;
Hermanns, Pia ;
Zabel, Bernhard ;
Dietz, Uwe H. .
DEVELOPMENTAL DYNAMICS, 2007, 236 (05) :1313-1318
[5]
Degenerative knee joint disease in mice lacking 3′-phosphoadenosine 5′-phosphosulfate synthetase 2 (Papss2) activity: a putative model of human PAPSS2 deficiency-associated arthrosis.[J].Alice F. Ford-Hutchinson;Zenobia Ali;Ruth A. Seerattan;David M.L. Cooper;Benedikt Hallgrímsson;Paul T. Salo;Frank R. Jirik.Osteoarthritis and Cartilage.2004, 5
[6]
Identification of sequence polymorphisms in two sulfation-related genes, PAPSS2 and SLC26A2, and an association analysis with knee osteoarthritis [J].
Ikeda, T ;
Mabuchi, A ;
Fukuda, A ;
Hiraoka, H ;
Kawakami, A ;
Yamamoto, S ;
Machida, H ;
Takatori, Y ;
Kawaguchi, H ;
Nakamura, K ;
Ikegawa, S .
JOURNAL OF HUMAN GENETICS, 2001, 46 (09) :538-543
[7]
Restricted and coordinated expression of β3-integrin and bone sialoprotein during cultured osteoblast differentiation [J].
Schneider, GB ;
Whitson, SW ;
Cooper, LF .
BONE, 1999, 24 (04) :321-327
[8]
THE VONKOSSA REACTION FOR CALCIUM DEPOSITS - SILVER LACTATE STAINING INCREASES SENSITIVITY AND REDUCES BACKGROUND [J].
RUNGBY, J ;
KASSEM, M ;
ERIKSEN, EF ;
DANSCHER, G .
HISTOCHEMICAL JOURNAL, 1993, 25 (06) :446-451
[9]
PAPSS2在大骨节病和骨关节炎疾病中作用机制的研究进展 [J].
赵全明 ;
王伟卓 ;
郭雄 .
国外医学(医学地理分册), 2012, 33 (02) :79-82