转运过程中Caco-2细胞对外源miR-133a摄取规律探究

The pattern of uptake for exogenous MiR-133a in Caco-2 cells in transport process

[目的]探究外源miR-133a在Caco-2细胞转运过程中细胞摄取的规律。[方法]建立Caco-2细胞模型以探究miR-133a在体内吸收情况;以Cy3标记的miR-133a对细胞摄取miR-133a的状态和过程进行探究;以细胞破碎液来评价被摄入的miR-133a在胞内的稳定性;通过细胞在不同温度下摄取miR-133a状态来评价细胞摄取miRNA途径。[结果]通过RT-q PCR检测到24 h内外源miR-133a能够以完整片段地形式被Caco-2细胞所转运,最大转运量占加入的总片段量的5.51×10^(-3)%;Caco-2细胞能够摄取外源miR-133a,24 h后达到饱和状态,吸收率达7.2%;被摄入的miR-133a主要分布于细胞质,且部分会围绕细胞核分布;摄入胞内的外源miR-133a在24 h后残留量占总量的0.04%;miR-133a的细胞摄取过程需要消耗能量。[结论]证明了Caco-2细胞能够主动摄取外源miR-133a,孵育24 h的吸收率为7.2%,且摄取为能量依赖过程。

[Objective] To explore the uptake for exogenous miR-133 a in Caco-2 cells. [Methods] The processes for transport of miR-133 a was explored by the Caco-2 cell monolayer model; The processes and patterns for uptake of miR-133 a in cells by Cy3 labeled miR-133 a were explored; It used cell lysate to imitate intracellular environment for evaluating intracellular stability of miR-133 a; The mechanism was evaluated by the uptake of miR-133 a at different temperatures. [Results]The results were showed by RT-q PCR that exogenous miR-133 a could be transported by Caco-2 cell model in a complete fragment; Caco-2 cells could uptake exogenous miR-133 a and the absorption rate of miR-133 a was 7. 2% when the uptake process reached saturation state after 24 hours; The exogenous miR-133 a was mainly distributed in cytoplasm,and some of it could be distributed around the nucleus; The residual amount of miR-133 a in cells accounted for 0. 04% of the total after 24 h; The uptake of miR-133 a was energy-dependent process. [Conclusion]Caco-2 cells can actively take in exogenous miR-133 a,the absorption rate of miR-133 a was 7. 2% after 24 h and the uptake is energy-dependent process.