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Low expression of microRNA-21 contributes to LPS-induced osteoblast cell apoptosis through up-regulation of OAS1

机译:microRNA-21的低表达通过上调OAS1促进LPS诱导的成骨细胞凋亡

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摘要

Lipopolysaccharide (LPS) is a critical component of the outer membrane of Gram-negative bacteria. Many cellular signals that are activated by Gram-negative bacteria are initiated by LPS. LPS triggers not only inflammatory responses, but also activates pro-apoptotic signals in a series of human cell types. However, there is relatively minimal data on the microRNA-dependent mechanism(s) of LPS-induced functional activity in osteoblast cells. CCK-8 assay and flow cytometry were used to measure cell viability and apoptosis, respectively. RT-PCR and western blot were performed to determine the mRNA and protein expression in osteoblast cells. In this study, we found that LPS triggered apoptosis in osteoblastic hFOB1.19 cells and induced a low expression of the miRNA-21. Furthermore, through the gene microarray technique, OAS1 was screened and later confirmed to be the target gene which was up-regulated in response to the low expression of miRNA-21. Knockdown of OAS1 by specific siRNAs significantly rescued the LPS-induced hFOB1.19 cell apoptosis. Our data suggest that LPS induces low expression of miRNA-21which consequently causes the up-regulation of the downstream gene OAS1 and eventually triggers apoptosis in hFOB1.19 cells. Knockdown of OAS1 rescues LPS-induced cell death and thus may be a promising therapeutic strategy for orthopedic diseases.
机译:脂多糖(LPS)是革兰氏阴性细菌外膜的重要组成部分。 LPS会引发革兰氏阴性细菌激活的许多细胞信号。 LPS不仅会触发炎症反应,还会激活一系列人类细胞类型中的促凋亡信号。然而,关于成骨细胞中LPS诱导的功能活性的microRNA依赖机制的数据相对较少。 CCK-8测定法和流式细胞仪分别用于测量细胞活力和凋亡。进行RT-PCR和western blot测定成骨细胞中的mRNA和蛋白表达。在这项研究中,我们发现LPS触发了成骨细胞hFOB1.19细胞的凋亡,并诱导了miRNA-21的低表达。此外,通过基因微阵列技术,筛选出OAS1,随后确认它是响应于miRNA-21的低表达而被上调的靶基因。通过特异性siRNA抑制OAS1可以显着挽救LPS诱导的hFOB1.19细胞凋亡。我们的数据表明,LPS诱导miRNA-21的低表达,从而引起下游基因OAS1的上调,并最终触发hFOB1.19细胞的凋亡。减少OAS1可以挽救LPS诱导的细胞死亡,因此可能是骨科疾病的有前途的治疗策略。

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