MicroRNA-335-5p inhibits osteoblast apoptosis induced by high glucose

Li,Jiling; Feng,Zhengping; Chen,Lixue; Wang,Xiaoju; Deng,Huacong

doi:10.3892/mmr.2016.4994

MicroRNA-335-5p inhibits osteoblast apoptosis induced by high glucose

Authors:
- Jiling Li
- Zhengping Feng
- Lixue Chen
- Xiaoju Wang
- Huacong Deng
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Affiliations: Department of Endocrinology, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, P.R. China, Laboratory Research Center, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, P.R. China
Published online on: March 16, 2016 https://doi.org/10.3892/mmr.2016.4994
Pages: 4108-4112

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Abstract

Diabetic osteoporosis represents a serious health condition with increasing incidence. Previous studies have shown that microRNA (miR)-335-5p is highly expressed in MC3T3-E1 osteoblasts and promotes their differentiation via downregulating the expression of dickkopf‑1 (DKK1). The present study investigated the effects of miR‑335‑5p on apoptosis of osteoblasts induced by high glucose (HG), as well as the underlying molecular mechanisms. MC3T3‑E1 osteoblasts were transfected with miR‑335‑5p mimics or control miR and cultured under HG conditions for seven days. Reverse‑transcription PCR and showed that, compared with the control group, the expression levels of miR‑335‑5p were significantly downregulated in the HG group. However, no significant differences were observed in the mRNA expression levels of DKK1 between these groups. Furthermore, flow cytometric analysis showed that the apoptotic rate was increased by >2‑fold in the HG group compared with that in the control group, while miR‑335‑5p overexpression significantly decreased the apoptotic rate in these model cells by ~40%. In addition, western blot analysis revealed that the protein expression levels of DKK1 and caspase‑3 were significantly elevated in the HG group, which was significantly inhibited by overexpression of miR‑335‑5p. These results may indicate that miR‑335‑5p overexpression inhibited HG‑induced apoptosis of MC3T3‑E1 osteoblasts through decreasing the protein expression levels of DKK1. The results of the present study suggested that miR‑335‑5p may represent a potential target for the treatment of diabetic osteoporosis.

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1	Wittrant Y, Gorin Y, Woodruff K, Horn D, Abboud HE, Mohan S and Abboud-Werner SL: High d(+)glucose concentration inhibits RANKL-induced osteoclastogenesis. Bone. 42:1122–1130. 2008. View Article : Google Scholar : PubMed/NCBI
2	Bartel DP: MicroRNAs: Genomics, biogenesis, mechanism and function. Cell. 116:281–297. 2004. View Article : Google Scholar : PubMed/NCBI
3	Mattick JS and Makunin IV: Non-coding RNA. Hum Mol Genet. 1:R17–R29. 2006. View Article : Google Scholar
4	Huntzinger E and Izaurralde E: Gene silencing by microRNAs: Contributions of translational repression and mRNA decay. Nat Rev Genet. 12:99–110. 2011. View Article : Google Scholar : PubMed/NCBI
5	Ciesla M, Skrzypek K, Kozakowska M, Loboda A, Jozkowicz A and Dulak J: MicroRNAs as biomarkers of disease onset. Anal Bioanal Chem. 401:2051–2061. 2011. View Article : Google Scholar : PubMed/NCBI
6	Song L and Tuan RS: MicroRNAs and cell differentiation in mammalian development. Birth Defects Res C Embryo Today. 78:140–149. 2006. View Article : Google Scholar : PubMed/NCBI
7	Davis BN and Hata A: Regulation of MicroRNA Biogenesis: A miRiad of mechanisms. Cell Commun Signal. 7:182009. View Article : Google Scholar : PubMed/NCBI
8	Stern-Ginossar N, Elefant N, Zimmermann A, Wolf DG, Saleh N, Biton M, Horwitz E, Prokocimer Z, Prichard M, Hahn G, et al: Host immune system gene targeting by a viral miRNA. Science. 317:376–381. 2007. View Article : Google Scholar : PubMed/NCBI
9	Shu M, Zheng X, Wu S, Lu H, Leng T, Zhu W, Zhou Y, Ou Y, Lin X, Lin Y, et al: Targeting oncogenic miR-335 inhibits growth and invasion of malignant astrocytoma cells. Mol Cancer. 10:592011. View Article : Google Scholar : PubMed/NCBI
10	Zhang J, Tu Q, Bonewald LF, He X, Stein G, Lian J and Chen J: Effects of miR-335-5p in modulating osteogenic differentiation by specifically downregulating Wnt antagonist DKK1. J Bone Miner Res. 26:1953–1963. 2011. View Article : Google Scholar : PubMed/NCBI
11	Hu Z, Wang B, Cao X, Du T and Zhang S: Prediction of micro-RNA related to change of mouse osteoblast function under simulated microgravity with bioinformatics method. Space Med Med Eng. 25:407–411. 2012.In Chinese.
12	Sato N, Yamabuki T, Takano A, Koinuma J, Aragaki M, Masuda K, Ishikawa N, Kohno N, Ito H, Miyamoto M, et al: Wnt inhibitor Dickkopf-1 as a target for passive cancer immunotherapy. Cancer Res. 70:5326–5336. 2010. View Article : Google Scholar : PubMed/NCBI
13	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods. 25:402–408. 2001. View Article : Google Scholar
14	Jackuliak P and Payer J: Osteoporosis, fractures, and diabetes. Int J Endocrinol. 2014:8206152014. View Article : Google Scholar : PubMed/NCBI
15	Marie PJ: Osteoblast dysfunctions in bone diseases: From cellular and molecular mechanisms to therapeutic strategies. Cell Mol Life Sci. 72:1347–1361. 2015. View Article : Google Scholar
16	Li H, Xie S, Liu M, Chen Z, Liu X, Wang L, Li D and Zhou Y: The clinical significance of downregulation of mir-124-3p, mir-146a-5p, mir-155-5p and mir-335-5p in gastric cancer tumorigenesis. Int J Oncol. 45:197–208. 2014.PubMed/NCBI
17	Cui XS, Sun SC, Kang YK and Kim NH: Involvement of microRNA-335-5p in cytoskeleton dynamics in mouse oocytes. Reprod Fertil Dev. 25:691–699. 2013. View Article : Google Scholar
18	Oger F, Gheeraert C, Mogilenko D, Benomar Y, Molendi-Coste O, Bouchaert E, Caron S, Dombrowicz D, Pattou F, Duez H, et al: Cell-specific dysregulation of microRNA expression in obese white adipose tissue. J Clin Endocrinol Metab. 99:2821–2833. 2014. View Article : Google Scholar : PubMed/NCBI
19	Zhang J, Zhu X, Cui J, Chen P, Wang S and Wang J: Differential expressions of microRNA between young and senescent endothelial cells. Chin Med J. 92:2205–2209. 2012.In Chinense.
20	Ertekin S, Yıldırım O, Dinç E, Ayaz L, Fidanci SB and Tamer L: Evaluation of circulating miRNAs in wet age-related macular degeneration. Mol Vis. 20:1057–1066. 2014.PubMed/NCBI
21	Wang XJ, Feng ZP, Deng HC, Jiang R and Du J: Effect of AP-1 on apoptosis of MC3T3-E1 osteoblast induced by high-glucose. Zhong Hua Gu Zhi Shu Song He Gu Kuang Yan Ji Bing Za Zhi. 7:258–262. 2014.In Chinese.
22	Li J, Sarosi I, Cattley RC, Pretorius J, Asuncion F, Grisanti M, Morony S, Adamu S, Geng Z, Qiu W, et al: Dkk1-mediated inhibition of Wnt signaling in bone results in osteopenia. Bone. 39:754–766. 2006. View Article : Google Scholar : PubMed/NCBI
23	Morvan F, Boulukos K, Clément-Lacroix P, Roman Roman S, Suc-Royer I, Vayssière B, Ammann P, Martin P, Pinho S, Pognonec P, et al: Deletion of a single allele of the Dkk1 gene leads to an increase in bone formation and bone mass. J Bone Miner Res. 21:934–945. 2006. View Article : Google Scholar : PubMed/NCBI
24	Qiang YW, Barlogie B, Rudikoff S and Shaughnessy JD Jr: Dkk1-induced inhibition of Wnt signaling in osteoblast differentiation is an underlying mechanism of bone loss in multiple myeloma. Bone. 42:669–680. 2008. View Article : Google Scholar : PubMed/NCBI
25	Wang LF, Bai D and Han XL: Progress in Dickkopf-1-mediated bone metabolism. J Clin Rehabil Tissue Eng Res. 17:337–341. 2013.In Chinese.