miRs在力学拉伸促进成肌细胞增殖过程中的作用
张马辉, 王永魁, 蒋兴禄, 余磊, 欧阳钧, 邱小忠, 王乐禹
中国临床解剖学杂志 ›› 2014, Vol. 32 ›› Issue (3) : 306-311.
miRs在力学拉伸促进成肌细胞增殖过程中的作用
The role of miRs in the process of promoting myoblast proliferation by mechanical stretch
目的 探讨3种miRs在力学拉伸促进C2C12成肌细胞增殖过程中的作用。 方法 周期性拉伸的各种力学条件通过 BioFlex 加载系统实现。 采用CCK-8检测不同拉伸频率对成肌细胞增殖的影响。对促增殖拉伸组和对照组进行高通量测序, 反转录定量 PCR(qRT-PCR) 验证高通量基因测序结果，并进行生物信息学分析。 结果 0.125 Hz拉伸组明显促进C2C12成肌细胞增殖（P<0.05），而0.25 Hz 和0.5 Hz相比于对照组的差异无统计学意义；高通量测序分析表明，11种miRs表达于对照组和0.125 Hz拉伸组，其中差异有统计学意义的为3种：mir-44，mir-36，mir-20；qRT-PCR证实这3种miRs的表达改变与测序结果一致；这3种miRs参与了多个信号通路的调控。 结论 高通量基因测序和生物信息学分析表明3种miRs在应力诱导的C2C12成肌细胞增殖过程中有重要作用。
Objective To explore the role of three kinds of miRs in the process of promoting myoblast proliferation by mechanical stretch. Methods Application of cyclic mechanical strain using the computer-controlled Flexcell Strain Unit cultured C2C12 cells. CCK-8 assay was applied to test myoblast proliferation in different conditions of mechanical stretch. The expression profiles of miRs in mechanical stretch groups and control groups were detected by high-throughput sequencing. The results were verified by reverse transcription quantitative PCR(qRT-PCR) and analyzed by bioinformatics． Results C2C12 cells in 0.125Hz stretching group was significantly observed with promoted proliferation of C2C12 myoblasts（P<0.05）, but no significant difference of cell proliferation were detected inthe 0.25 Hz and 0.5Hz groups compared to that in the control group; Using high-throughput sequencing technology, we found 11 kinds of miRs which were co-expressed and differentially expressed in the two groups (Control group and 0.125 Hz stretching group); the difference was statistically significant in the expression of three miRs: mir-44 , mir-36 and mir-20. These three miRs expression was verified by qRT-PCR, and the results were consistent with the sequencing. These three miRs were involved in several signaling pathways. Conclusions High-throughput genome sequencing and bioinformatics analysis show that three kinds of miRs play an important role in the proliferation of C2C12 myoblast.
MiRs / Mechanical stretch / Cellular proliferation / High-throughput sequencing
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