摘要
目的探讨Bcl-2和Bak基因沉默对MG-63细胞凋亡、成骨活动以及钙离子浓度的影响。方法构建沉默Bcl-2和BaK表达的腺病毒载体,scramble RNA载体转染MG-63细胞,采用MTT检测转染后细胞存活率的情况,并应用ALP观察转染后细胞成骨活动情况以及流式细胞术检测钙离子的浓度间接测定细胞凋亡情况。结果 Bcl-2基因沉默后48 h相对对照组,细胞存活率降低,成骨活动减少,钙离子浓度升高;BaK基因沉默后细胞增存活率升高,成骨活动增加,细胞内钙离子浓度降低,而Bcl-2+BaK联合沉默组与对照组在各观察指标上差异无显著性。结论 Bcl-2和BaK基因沉默对人成骨肉瘤MG-63细胞凋亡,钙离子浓度变化以及成骨活动产生影响,表明其可能与骨质疏松的发病有密切的关联。
Objective To explore the effects of Bcl-2 and BaK gene silencing on cell apoptosis, osteogenesis activity and free Ca^2＋ concentration of MG-63 cell lines. Methods The siRNA sequences targeted Bcl-2 and BaK respectively were designed ; Bcl-2 and BaK silencing adenovirus vector scramble RNA vector and empty vector were constructed to transfect MG-63 cell lines. MTT method was used to examine cell viability ; ALP and flow cytometry were conducted to observe osteogenesis activity and free Ca^2＋ concentration. Results Bcl-2 gene silencing decreased cell viability, reduced osteogenesis activity and increased free Ca^2＋ concentration when compared with controls but BaK gene silencing had the opposite effects. The effect of Bcl-2＋BaK gene silencing on cell was similar to the empty control. Conclusions Cell apoptosis, osteogenesis activity and free Ca2~ concentration of MG- 63 change following Bcl-2 and BaK gene silencing, implicating their roles in osteoporosis.
出处
《实用医学杂志》
CAS
北大核心
2017年第17期2813-2816,共4页
The Journal of Practical Medicine
基金
国家自然科学基金(编号:81674004
81373653)
广东省科技计划项目(编号:2016A020216024)
作者简介
通信作者：黄宏兴E—mail：hxhortho@126．com
