PRELIMINARY ANALYSIS OF DIFFERENTIALLY EXPRESSED GENES IN STEROID-INDUCED OSTEONECROSIS OF FEMORAL HEAD BY GENE MICROARRAY_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

 CHENGXiangguo , HUBin , SHENGJiagen , ZHANGChangqing

Department of Orthopedics, No.6 People's Hospital Affiliated to Shanghai Jiaotong University, Shanghai, 200233, P. R. China;

Corresponding author：

CHENGXiangguo, Email: xgcheng@sjtu.edu.cn

Keywords：

Steroid-induced osteonecrosis of the femoral head; Gene microarray; Differentially expressed gene

DOI：

10.7507/1002-1892.20140131

Video：

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Objective To screen for the differentially expressed genes in steroid-induced osteonecrosis of the femoral head (ONFH) by gene microarray. Methods The femoral head tissue of ONFH was harvested from 3 patients with steroid-induced ONFH, aged 25, 31, and 38 years, respectively. Normal tissue was harvested from a 26-year-old male remains contributor. HE staining of the specimens was performed for observing the histology manifestation; the total RNA was extracted for measuring the purity; cDNA probe was synthesized by reverse transcription, and then were hybridized as the cDNA microarray for scanning of fluorescent signals and differentially expressed genes in the tissues. Results HE staining of normal tissue showed complete unit composed of lamellar bone, continuous and complete lamellar bone with a concentric arrangement around blood vessels, and normal bone cells in the trabecular bone lacuna. In ONFH tissue, adipose tissue increased in the medullary cavity, with increased fat cells filling in the medullary cavity and extruding capillary, and with decreased bone cells in the bone trabecula, which had deeply-stained nuclear chromatin, pyknotic or cracking nucleus, and even bone cells disappeared in the part of the bone lacuna, and trabecular bone became thin, sparse, interrupt, reduced area in visual field/unit. Total RNA extraction electrophoretogram displayed clear bands of 28S and 18S, and the brightness ratio of the 28S:18S was 2:1, indicating good total RNA quality. And 44 genes were differentially expressed, and there were 28 up-regulated genes and 16 down-regulated genes, including cell/organism defense genes, cell structure/motility genes, cell division genes, cell signaling/cell communication genes, cell metabolism genes, gene/protein expression genes, and unclassified genes. Conclusion The analysis of the gene expression profile of steroid-induced ONFH can provide evidence for the pathogenesis of ONFH.

Citation： CHENGXiangguo, HUBin, SHENGJiagen, ZHANGChangqing. PRELIMINARY ANALYSIS OF DIFFERENTIALLY EXPRESSED GENES IN STEROID-INDUCED OSTEONECROSIS OF FEMORAL HEAD BY GENE MICROARRAY. Chinese Journal of Reparative and Reconstructive Surgery, 2014, 28(5): 586-590. doi: 10.7507/1002-1892.20140131 Copy

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2.	Issa K, Pivec R, Kapadia BH, et al. Osteonecrosis of the femoral head:the total hip replacement solution. Bone Joint J, 2013, 95-B(11 Suppl A):46-50.
3.	Zhao FC, Li ZR, Guo KJ. Clinical analysis of osteonecrosis of the femoral head induced by steroids. Orthop Surg, 2012, 4(1):28-34.
4.	Jones JP Jr. Concepts of etiology and early pathogenesis of osteonecrosis. Instr Course Lect, 1994, 43:499-512.
5.	Nakamura J, Ohtori S, Watanabe A, et al. Recovery of the blood flow around the femoral head during early corticosteroid therapy:dynamic magnetic resonance imaging in systemic lupus erythematosus patients. Lupus, 2012, 21(3):264-270.
6.	Paydas S, Balal M, Demir E, et al. Avascular osteonecrosis and accompanying anemia, leucocytosis, and decreased bone mineral density in renal transplant recipients. Transplant Proc, 2011, 43(3):863-866.
7.	Motomura G, Yamamoto T, Miyanishi K, et al. Risk factors for developing osteonecrosis after prophylaxis in steroid-treated rabbits. J Rheumatol, 2008, 35(12):2391-2394.
8.	Chauhan V, Ranganna KM, Chauhan N, et al. Bone disease in organ transplant patients:pathogenesis and management. Postgrad Med, 2012, 124(3):80-90.
9.	Erken HY, Ofluoglu O, Aktas M, et al. Effect of pentoxifylline on histopathological changes in steroid-induced osteonecrosis of femoral head:experimental study in chicken. Int Orthop, 2012, 36(7):1523-1528.
10.	Ragni E, Montemurro T, Montelatici E, et al. Differential microRNA signature of human mesenchymal stem cells from different sources reveals an "environmental-niche memory" for bone marrow stem cells. Exp Cell Res, 2013, 319(10):1562-1574.
11.	Weinstein RS. Glucocorticoid-induced osteoporosis and osteonecrosis. Endocrinol Metab Clin North Am, 2012, 41(3):595-611.
12.	Mehan MR, Ostroff R, Wilcox SK, et al. Highly multiplexed proteomic platform for biomarker discovery, diagnostics, and therapeutics. Adv Exp Med Biol, 2013, 735:283-300.
13.	Kerachian MA, Séguin C, Harvey EJ. Glucocorticoids in osteonecrosis of the femoral head:a new understanding of the mechanisms of action. J Steroid Biochem Mol Biol, 2009, 114(3-5):121-128.
14.	O'Brien MA, Costin BN, Miles MF. Using genome-wide expression profiling to define gene networks relevant to the study of complex traits:from RNA integrity to network topology. Int Rev Neurobiol, 2012, 104:91-133.
15.	Zehner M, Burgdorf S. Regulation of antigen transport into the cytosol for cross-presentation by ubiquitination of the mannose receptor. Mol Immunol, 2013, 55(2):146-148.
16.	Morris CR, Stanton MJ, Manthey KC, et al. A knockout of the Tsg101 gene leads to decreased expression of ErbB receptor tyrosine kinases and induction of autophagy prior to cell death. PLoS One, 2012, 7(3):e34308.
17.	Gruenberg J. Viruses and endosome membrane dynamics. Curr Opin Cell Biol, 2009, 21(4):582-588.
18.	Wagner SA, Beli P, Weinert BT, et al. A proteome-wide, quantitative survey of in vivo ubiquitylation sites reveals widespread regulatory roles. Mol Cell Proteomics, 2011, 10(10):M111.
19.	Ma XR, Edmund Sim UH, Pauline B, et al. Overexpression of WNT2 and TSG101 genes in colorectal carcinoma. Trop Biomed, 2008, 25(1):46-57.
20.	Shen J, Abel EL, Riggs PK, et al. Proteomic and pathway analyses reveal a network of inflammatory genes associated with differences in skin tumor promotion susceptibility in DBA/2 and C57BL/6 mice. Carcinogenesis, 2012, 33(11):2208-2219.
21.	杨雨润, 娄晋宁, 李子荣, 等. 糖皮质激素对骨髓微血管内皮细胞活性氧代谢影响的实验研究. 中国修复重建外科杂志, 2011, 25(5):533-537.
22.	Winbanks CE, Chen JL, Qian H, et al. The bone morphogenetic protein axis is a positive regulator of skeletal muscle mass. J Cell Biol, 2013, 203(2):345-357.
23.	Grönroos E, Kingston IJ, Ramachandran A, et al. Transforming growth factor β inhibits bone morphogenetic protein-induced transcription through novel phosphorylated Smad1/5-Smad3 complexes. Mol Cell Biol, 2012, 32(14):2904-2916.
24.	Yang J, Li X, Al-Lamki RS, et al. Smad-dependent and smad-independent induction of id1 by prostacyclin analogues inhibits proliferation of pulmonary artery smooth muscle cells in vitro and in vivo. Circ Res, 2010, 107(2):252-262.
25.	Emes Y, Aybar B, Vural P, et al. Effects of bone morphogenetic proteins on osteoblast cells:vascular endothelial growth factor, calcium, inorganic phosphate, and nitric oxide levels. Implant Dent, 2010, 19(5):419-427.
26.	Tang Y, Yang X, Friesel RE, et al. Mechanisms of TGF-β-induced differentiation in human vascular smooth muscle cells. J Vasc Res, 2011, 48(6):485-494.

1. Wang CJ, Huang CC, Wang JW, et al. Long-term results of extracorporeal shockwave therapy and core decompression in osteonecrosis of the femoral head with eight-to nine-year follow-up. Biomed J, 2012, 35(6):481-485.
2. Issa K, Pivec R, Kapadia BH, et al. Osteonecrosis of the femoral head:the total hip replacement solution. Bone Joint J, 2013, 95-B(11 Suppl A):46-50.
3. Zhao FC, Li ZR, Guo KJ. Clinical analysis of osteonecrosis of the femoral head induced by steroids. Orthop Surg, 2012, 4(1):28-34.
4. Jones JP Jr. Concepts of etiology and early pathogenesis of osteonecrosis. Instr Course Lect, 1994, 43:499-512.
5. Nakamura J, Ohtori S, Watanabe A, et al. Recovery of the blood flow around the femoral head during early corticosteroid therapy:dynamic magnetic resonance imaging in systemic lupus erythematosus patients. Lupus, 2012, 21(3):264-270.
6. Paydas S, Balal M, Demir E, et al. Avascular osteonecrosis and accompanying anemia, leucocytosis, and decreased bone mineral density in renal transplant recipients. Transplant Proc, 2011, 43(3):863-866.
7. Motomura G, Yamamoto T, Miyanishi K, et al. Risk factors for developing osteonecrosis after prophylaxis in steroid-treated rabbits. J Rheumatol, 2008, 35(12):2391-2394.
8. Chauhan V, Ranganna KM, Chauhan N, et al. Bone disease in organ transplant patients:pathogenesis and management. Postgrad Med, 2012, 124(3):80-90.
9. Erken HY, Ofluoglu O, Aktas M, et al. Effect of pentoxifylline on histopathological changes in steroid-induced osteonecrosis of femoral head:experimental study in chicken. Int Orthop, 2012, 36(7):1523-1528.
10. Ragni E, Montemurro T, Montelatici E, et al. Differential microRNA signature of human mesenchymal stem cells from different sources reveals an "environmental-niche memory" for bone marrow stem cells. Exp Cell Res, 2013, 319(10):1562-1574.
11. Weinstein RS. Glucocorticoid-induced osteoporosis and osteonecrosis. Endocrinol Metab Clin North Am, 2012, 41(3):595-611.
12. Mehan MR, Ostroff R, Wilcox SK, et al. Highly multiplexed proteomic platform for biomarker discovery, diagnostics, and therapeutics. Adv Exp Med Biol, 2013, 735:283-300.
13. Kerachian MA, Séguin C, Harvey EJ. Glucocorticoids in osteonecrosis of the femoral head:a new understanding of the mechanisms of action. J Steroid Biochem Mol Biol, 2009, 114(3-5):121-128.
14. O'Brien MA, Costin BN, Miles MF. Using genome-wide expression profiling to define gene networks relevant to the study of complex traits:from RNA integrity to network topology. Int Rev Neurobiol, 2012, 104:91-133.
15. Zehner M, Burgdorf S. Regulation of antigen transport into the cytosol for cross-presentation by ubiquitination of the mannose receptor. Mol Immunol, 2013, 55(2):146-148.
16. Morris CR, Stanton MJ, Manthey KC, et al. A knockout of the Tsg101 gene leads to decreased expression of ErbB receptor tyrosine kinases and induction of autophagy prior to cell death. PLoS One, 2012, 7(3):e34308.
17. Gruenberg J. Viruses and endosome membrane dynamics. Curr Opin Cell Biol, 2009, 21(4):582-588.
18. Wagner SA, Beli P, Weinert BT, et al. A proteome-wide, quantitative survey of in vivo ubiquitylation sites reveals widespread regulatory roles. Mol Cell Proteomics, 2011, 10(10):M111.
19. Ma XR, Edmund Sim UH, Pauline B, et al. Overexpression of WNT2 and TSG101 genes in colorectal carcinoma. Trop Biomed, 2008, 25(1):46-57.
20. Shen J, Abel EL, Riggs PK, et al. Proteomic and pathway analyses reveal a network of inflammatory genes associated with differences in skin tumor promotion susceptibility in DBA/2 and C57BL/6 mice. Carcinogenesis, 2012, 33(11):2208-2219.
21. 杨雨润, 娄晋宁, 李子荣, 等. 糖皮质激素对骨髓微血管内皮细胞活性氧代谢影响的实验研究. 中国修复重建外科杂志, 2011, 25(5):533-537.
22. Winbanks CE, Chen JL, Qian H, et al. The bone morphogenetic protein axis is a positive regulator of skeletal muscle mass. J Cell Biol, 2013, 203(2):345-357.
23. Grönroos E, Kingston IJ, Ramachandran A, et al. Transforming growth factor β inhibits bone morphogenetic protein-induced transcription through novel phosphorylated Smad1/5-Smad3 complexes. Mol Cell Biol, 2012, 32(14):2904-2916.
24. Yang J, Li X, Al-Lamki RS, et al. Smad-dependent and smad-independent induction of id1 by prostacyclin analogues inhibits proliferation of pulmonary artery smooth muscle cells in vitro and in vivo. Circ Res, 2010, 107(2):252-262.
25. Emes Y, Aybar B, Vural P, et al. Effects of bone morphogenetic proteins on osteoblast cells:vascular endothelial growth factor, calcium, inorganic phosphate, and nitric oxide levels. Implant Dent, 2010, 19(5):419-427.
26. Tang Y, Yang X, Friesel RE, et al. Mechanisms of TGF-β-induced differentiation in human vascular smooth muscle cells. J Vasc Res, 2011, 48(6):485-494.

Chinese Journal of Reparative and Reconstructive Surgery

PRELIMINARY ANALYSIS OF DIFFERENTIALLY EXPRESSED GENES IN STEROID-INDUCED OSTEONECROSIS OF FEMORAL HEAD BY GENE MICROARRAY

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