Bioinformatics analysis and identification of circular RNAs promoting the osteogenic differentiation of human bone marrow mesenchymal stem cells on titanium treated by surface mechanical attrition
Bioinformatics analysis and identification of circular RNAs promoting the osteogenic differentiation of human bone marrow mesenchymal stem cells on titanium treated by surface mechanical attrition
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DOI：
10.7717/peerj.9292
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发表时间：
2020-07
期刊：
影响因子：
2.7
通讯作者：
Shanshan Zhu;Yuhe Zhu;Zhenbo Wang;Chen Liang;Nanjue Cao;Ming Yan;Fei Gao;Jie Liu;Wei Wang-W
中科院分区：
文献类型：
--
作者：
Shanshan Zhu;Yuhe Zhu;Zhenbo Wang;Chen Liang;Nanjue Cao;Ming Yan;Fei Gao;Jie Liu;Wei Wang-W
背景分析和鉴定表面机械研磨处理（SMAT）促进人骨髓间充质干细胞（hBMSCs）在钛表面成骨分化的环状RNA（circRNA）。方法实验材料为SMAT纯钛，对照材料为退火纯钛。采用CCK-8法检测hBMSCs的增殖情况，碱性磷酸酶（ALP）活性及茜素红染色法检测hBMSCs在样品表面的成骨分化情况。利用生物信息学预测软件miwalk3.0，通过预测成骨相关信使RNA（mRNAs）和microRNA（miRNAs）的circRNA，构建竞争性内源性RNA（ceRNA）网络。通过实时定量PCR（QRT-PCR）选择并分析位于网络关键位置的circRNA。结果与退火纯钛相比，SMAT纯钛能促进hBMSCs的增殖和成骨分化。预测的circRNA总数为51。其中，30个circRNA和8个miRNAs构成了6个ceRNA网络。选择Circ-LTBP 2进行验证。QRT-PCR结果显示，与对照组相比，实验组hsa_circ_0032599、hsa_circ_0032600和hsa_circ_0032601的表达水平上调; hsa_circ_0032600差异表达最明显，差异有统计学意义，变化倍数（FC）= 4.25 ± 1.60，p值（p）< 0.05。
Background To analyze and identify the circular RNAs (circRNAs) involved in promoting the osteogenic differentiation of human bone mesenchymal stem cells (hBMSCs) on titanium by surface mechanical attrition treatment (SMAT). Methods The experimental material was SMAT titanium and the control material was annealed titanium. Cell Counting Kits-8 (CCK-8) was used to detect the proliferation of hBMSCs, and alkaline phosphatase (ALP) activity and alizarin red staining were used to detect the osteogenic differentiation of hBMSCs on the sample surfaces. The bioinformatics prediction software miwalk3.0 was used to construct competing endogenous RNA (ceRNA) networks by predicting circRNAs with osteogenesis-related messenger RNAs (mRNAs) and microRNAs (miRNAs). The circRNAs located at the key positions in the networks were selected and analyzed by quantitative real-time PCR (QRT-PCR). Results Compared with annealed titanium, SMAT titanium could promote the proliferation and osteogenic differentiation of hBMSCs. The total number of predicted circRNAs was 51. Among these, 30 circRNAs and 8 miRNAs constituted 6 ceRNA networks. Circ-LTBP2 was selected for verification. QRT-PCR results showed that the expression levels of hsa_circ_0032599, hsa_circ_0032600 and hsa_circ_0032601 were upregulated in the experimental group compared with those in the control group; the differential expression of hsa_circ_0032600 was the most obvious and statistically significant, with a fold change (FC) = 4.25 ± 1.60, p-values (p) < 0.05.