Adenovirus-mediated RNA interference against core binding factor alpha 1 inhibits the hypertrophic differentiation of chondrocytes

doi:10.3969/j.issn.2095-4344.2015.02.005

Abstract

Abstract:

BACKGROUND: Hypertrophic differentiation of chondrocytes is the sign of starting endochondral ossification, and it is also an essential step in endochondral ossification, which is a cascade reaction and difficult to be blocked once started. The end result is the formation of bone structure. RNA interference is a post-transcriptional gene silencing. Relevant studies have shown that the use of RNA interference to block the expression of core binding factor α1 (Cbfα1) can effectively inhibit the formation of heterotopic ossification.
OBJECTIVE: To use RNA intereference technology to suppress Cbfα1 expression so as to achieve the purpose of blocking the hypertrophic diferentiation of chondrocytes.
METHODs: We constructed an adenovirus containing siRNA against Cbfα1 (Ad-Cbfα1-siRNA). Retinoic acid and interleukin-1α were used to induce hypertrophic differetiation of chondrocytes, and then Ad-Cbfα1-siRNA was utilized to inhibit the hypertrophic differentiation of chondrocytes. Immunohistochemistry method was used to analyze the expression of Cbfα1.
RESULTS AND CONCLUSION: After induction with retinoic acid and interleukin-1α, the chondrocytes in the negative control virus group appeared to have hypertrophy and the expression of Cbfα1 was positive. In the Ad-Cbα1-siRNA group, the expression of Cbfα1 was negative. These findings suggest that the inhibition of Cbfα1 by RNA interference can be a powerful way to prevent the hypertrophic differentiation of chondrocytes.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

全文链接：

Key words: Chondrocytes, Core Binding Factors, RNA Interference, Adenoviridae

CLC Number:

R318

Gao Bo, Xing Rong, Kong Qing-quan, Xiang Zhou, Yang Jing, Cai Jia-qin, Huang Yi-zhou,. Adenovirus-mediated RNA interference against core binding factor alpha 1 inhibits the hypertrophic differentiation of chondrocytes[J]. Chinese Journal of Tissue Engineering Research, 2015, 19(2): 187-191.

Figures/Tables 3

References

[1] van der Kraan PM, van den Berg WB. Chondrocyte hypertrophy and osteoarthritis:role in initiation and progression of cartilage degeneration? Osteoarthritis Cartilage. 2012;20(3):223-232.
[2] Lin L, Chen L, Wang H, et al. Adenovirus-mediated transfer of siRNA against Runx2/Cbfa1 inhibits the formation of heterotopic ossification in animal model. Biochem Biophys Res Commun.2006;349(2):564-572.
[3] 杨志明. 组织工程基础与临床[M].成都:四川科学技术出版社，2000:151-152.
[4] Glasson SS, Askew R, Sheppard B, et al. Deletion of active ADAMTS5 prevents cartilage degradation in a murine model of osteoarthritis. Nature. 2005;434(7033):644-648.
[5] Lories RJ,Luyten FP. The bone-cartilage unit in osteoarthritis. Nat Rev Rheummatol. 2011;7(1):43-49.
[6] Inada M, Yasui T, Nomura S, et al. Maturational disturbance of chondrocytes in Cbfa1-deficient mice. Dev Dyn. 1999;214(4): 279-290.
[7] Guo S, Kemphues KJ. par-1, a gene required for establishing polarity in C.elegan embryos, encodes a putative Ser/Thr kinase that is asymmetrically distributed. Cell. 1995; 81(4):611-620.
[8] Fire A, Xu SQ, Montgomery MK, et al. Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature.1998,391(6669):806-811.
[9] Kim DH, Rossi JJ. Strategies for silencing human disease using RNA interference. Nat Rev Genet. 2007;8(3):173-184.
[10] Schiffelers RM, Xu J, Storm G, et al. Effects of treatment with small interfering RNA on joint inflammation in mice with collagen-induced arthritis. Arthritis Rheum. 2005,52(4): 1314-1318.
[11] Xue T, Mao Z,Lin L, et al. Non-virus-mediated transfer of siRNAs against Runx2 and Smad4 inhibit heterotopic ossification in rats. Gene Ther.2010;17(3):370-379.
[12] Owen LA, Lessnick SL. Identification of target genes in their native cellular context: an analysis of EWS/FLI in Ewing’s sarcoma. Cell Cycl.2006;5(18):2049-2053.
[13] Lianxu C, Hongti J, Changlong Y. NF-ΚBp65-specific siRNA inhibits expression of genes of COX-2, NOS-2 and MMP-9 in rat IL-1β-induced and TNF-α-induced chondrocytes. Osteoathritis Cartilage. 2006;14(4):367-376.
[14] Kamekura S, Kawasaki Y, Hoshi K, et al. Contribution of runt-related transcription factor 2 to the pathogenesis of oeteoarthritis in mice after induction of knee joint insatability. Arthritis Rheum. 2006;54(8):2462-2470.
[15] Gauci SJ, Golub SB, Tutolo L, et al. Modulating chondrocyte hypertrophy in growth plate and osteoarthritic cartilage. J Musculoskelet Neuronal Interact. 2008;8(4):308-310.
[16] Tchetina EV, Kobayashi M, Yasuda T, et al. Chondrocyte hypertrophy can be induced by a cryptic sequence of type II collagen and is accompanied by the induction of MMP-13 and collagenase activity: Implications for development and arthritis. Matrix Biol. 2007;26(4) :247-258.
[17] Morita K, Miyamoto T, Fujita N,et al. Reactive oxygen species induce chondrocyte hypertrophy in endochondral ossification. JEM. 2007;204(7):1613-1623.
[18] Merz D, Liu R, Johnson K, et al. IL-8/CXCL8 and growth-related oncogene α/CXCL1 induce chondrocyte hypertrophic differentiation. J Immunol. 2003;171(8): 4406-4415.
[19] Guo XH, Mark KK,Taketo MM, et al. The Wnt/β-Catenin pathway interacts differentially with PTHrP signaling to control chondrocyte hypertrophy and final maturation. PLoS ONE. 2009;4(6):e6067.
[20] Komori T, Yagi H, Nomura S, et al. Targeted disruption of cbfa1 results in a complete lack of bone formation owing tomaturational arrest of osteoblasts. Cell. 1997;89(5): 755-764.
[21] Enomoto H, Enomoto-Iwamoto M, Iwamoto M,et al. Cbfα1 is a positive regulatory factor in chondrocyte maturation. J Biol Chem. 2000;275(12):8695-8702.
[22] Zheng LH, Baek HJ, Karsenty G, et al. Filamin B represses chondrocyte hypertrophy in a Runx2/Smad3-dependent manner. J Cell Biol. 2007;178(1):121-128.