SelW mRNA下调对肌细胞状态影响的FCM评估

Assessment of Cell Conditions by FCM after SelW mRNA Down-Regulation

运用RNA干扰技术,在转录后阶段实现肌细胞中SelW mRNA的定量下调,并利用流式细胞术客观地区分、计数不同状态的细胞,考察C2C12细胞中SelW mRNA的定量下调对细胞状态的影响。结果显示:当siR-NA剂量为2.0,1.5,1.2μg/g时,细胞脱落、死亡;当siRNA剂量为1.0,0.5,0.25μg/g时,凋亡率分别为32.5%,12.4%,6.58%。在干预评估试验中,阴性对照组与空白对照组差异显著(P<0.01)。细胞状态随SelW mRNA的下调存在着阶段性变化,凋亡率与mRNA的降低量之间的关系为非线性正相关,证明SelW对肌细胞的健康是必须的。显著的凋亡率表明SelW对肌细胞的功能重要性的机制是十分复杂的,尚需进一步研究。

The influence of SelW mRNA down-regulation on the status of C2C12 cell was assayed. RNAi was performed to carry out SleW mRNA down-regulation after transcription. Cells in different status were distinguished and recorded objectively by Flow Cytometry. When 2.0, 1.5, 1.2 μg/g siRNA were applied to down-regulate SelW mRNA, death and detachment of cell had been found. While 1.0, 0.5, 0.25 μg/g siRNA were used, apoptosis rates of 32.5%, 12.4% and 6.58% were obtained. And in experimental interference assay, there was a significant difference between negative control and blank group （ P 〈 0.01 ）. Conclusion： the changes of cell status were coordinate to the degrees of SelW mRNA downregulation ; apoptotic rate changing followed the dropping of mRNA, which was non-linearity positive correlation. These illustrated SelW was essential to the health of skeletal muscle cell. The significant apoptosis rate demonstrated the mechanism of how SelW plays its role was very complicated, and further research is needed.