摘要
目的:观察机械牵张应力作用下人成骨肉瘤细胞MG-63中丝切蛋白(Cofilin)的表达。方法:将MG-63细胞分为加力组与对照组(不加力),加力组采用Flexcell牵张应力加载系统,用12%变形率的应力值进行力学刺激1、4、8、12、24 h,用免疫荧光染色观察Cofilin的变化,RT-PCR检测Cofilin mRNA水平含量的变化,Western-Blot测定细胞内Cofilin蛋白含量。结果:MG-63细胞受应力刺激后1、4 h细胞内Cofilin染色逐渐增强,8、12、24 h细胞染色逐渐减弱,但仍高于对照组。应力作用下Cofilin mRNA水平无明显变化(P>0.05)。Western Blot显示,加载后1 h细胞内Cofilin含量增高(P<0.05),4 h时达到高峰(P<0.05),8、12、24 h后Cofilin含量逐渐减少,但仍高于对照组(P<0.05)。结论:机械牵张应力刺激可明显上调Cofi-lin在MG-63细胞中表达,且具有时间依赖性,提示其可能在成骨细胞力学信号转导过程中发挥重要作用。
Objective： To investigate the effects of mechanical strain on cofilin expression in MG-63 osteoblastic cells. Methods： Mechanical strain at 12% was applied to MG-63 ceils for 0（ control group）, 1,4,8,12 and 24 hours respectively in vitro. Cofilin mR- NA expression in the ceils was examined by RT-PCR, cofilin protein expression was examined by immunofluoresence assay and West- ern Blotting. Results： The fluorescence intensity of the cells was enhanced from 1 to 4 hours after strain treatment, then decreased from 8 to 24 hours, mRNA was not significantly changed after stain treatment（P 〉 0.05 ）. Western blotting showed that cofilin expres- sion level in MG-63 cells was gradually increased within the first 4 hours of mechanical strain application （ P 〈 0.05 ）, then decreased in 8, 12 and 24 hours （P 〈0.05 ） ,but still higher than that in the control ceils. Conclusion： Mechanical strain elongation regulates cofilin expression of MG-63 cells. Cofilin may plays an important role in cell signal transduction of mechanical strain.
出处
《实用口腔医学杂志》
CAS
CSCD
北大核心
2013年第3期308-312,共5页
Journal of Practical Stomatology
基金
国家自然科学基金资助(编号:31070836)
青年科学基金(编号:31200706)
