摘要
背景:机械牵引力能够影响MC3T3-E1细胞的增殖分化过程,并引起细胞内miR-132-3p的差异表达。然而,牵引力是否通过调控miR-132-3p的表达来影响成骨细胞增殖分化仍需进一步研究。目的:明确12%牵引力作用下MC3T3-E1细胞中成骨分化标志因子及miR-132-3p表达变化,并进一步探讨miR-132-3p对细胞增殖分化的影响。方法:MC3T3-E1细胞分别加载0%,12%牵张应力,检测应力加载后碱性磷酸酶活性、骨钙蛋白及miR-132-3pmRNA的表达水平;细胞内瞬时转染miR-132-3p模拟物及其阴性对照,qRT-PCR检测转染后碱性磷酸酶、骨钙蛋白、Runt标志转录因子2mRNA的表达,CCK-8法检测miR-132-3p对细胞增殖能力的影响。结果与结论:①12%牵张应力作用下,MC3T3-E1细胞中碱性磷酸酶活性、骨钙蛋白mRNA表达水平下调(P<0.01),miR-132-3p表达水平显著升高(P<0.05);②细胞内转染miR-132-3p后,miR-132-3p模拟物组成骨分化标志因子碱性磷酸酶、骨钙蛋白、Runt标志转录因子2mRNA表达水平显著降低(P<0.05);③相比于阴性对照组,miR-132-3p模拟物转染24,48,72h后细胞增殖能力明显降低(P<0.001),且在转染48h后降低最明显;④结果说明12%周期性循环牵张应力能够通过过表达miR-132-3p负向调节MC3T3-E1细胞的增殖和成骨分化。
BACKGROUND:Mechanical stress can influence the proliferation and differentiation of MC3T3-E1 cells and trigger differential expression of miR-132-3p.However,further research is warranted concerning whether tensile stress can influence the proliferation and differentiation of osteoblasts by regulating miR-132-3p.OBJECTIVE:To determine the expression of osteogenic differentiation markers and miR-132-3p in MC3T3-E1 cells under 12%cyclic stretch and to explore the effect of miR-132-3p on cell proliferation and differentiation.METHODS:MC3T3-E1 cells were loaded with 0%and 12%tensile stress,and alkaline phosphatase activity,osteocalcin mRNA and miR-132-3p expression levels were detected.MC3T3-E1 cells were transiently transfected with miR-132-3p mimics and a negative control transfection group was set up.The expression of alkaline phosphatase,osteocalcin and Runx2 mRNA in transfected cells were detected by qRT-PCR,and the effect of miR-132-3p on cell proliferation were detected by cell counting kit-8 assay.RESULTS AND CONCLUSION:The alkaline phosphatase activity and osteocalcin mRNA expression were down-regulated in MC3T3-E1 cells under 12%stretch stress(P<0.01),and the expression of miR-132-3p was significantly increased(P<0.05).QRT-PCR results showed the expression levels of osteogenic differentiation markers alkaline phosphatase activity,osteocalcin,and Runx2 mRNA in miR-132-3p mimics group were significantly decreased after intracellular transfection of miR-132-3p(P<0.05).Compared with the negative control transfection group,the cell proliferation in the miR-132-3p mimic group was decreased at 24,48,and 72 hours after transfection(P<0.001),and the most obvious reduction was observed after 48-hour transfection.These findings indicate that 12%cyclic tensile stress can negatively regulate the proliferation and differentiation ability of MC3T3-E1 cells by overexpressing miR-132-3p.
作者
孙芬
刘名燕
刘铭
孙心旖
冯云霞
Sun Fen;Liu Mingyan;Liu Ming;Sun Xinyi;Feng Yunxia(Shanxi Medical University School and Hospital of Stomatology,Taiyuan 030000,Shanxi Province,China;Dental Doctor,Suzhou 215000,Jiangsu Province,China;School of Basic Medical Science,Shanxi Medical University,Taiyuan 030000,Shanxi Province,China)
出处
《中国组织工程研究》
CAS
北大核心
2020年第1期59-64,共6页
Chinese Journal of Tissue Engineering Research
基金
国家自然科学基金(81400566),项目负责人:刘名燕
