miRs在力学拉伸促进成肌细胞增殖过程中的作用
收稿日期: 2013-11-20
网络出版日期: 2014-06-04
基金资助
国家自然科学基金（31100700）
The role of miRs in the process of promoting myoblast proliferation by mechanical stretch
Received date: 2013-11-20
Online published: 2014-06-04
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。
张马辉 , 王永魁 , 蒋兴禄 , 余磊 , 欧阳钧 , 邱小忠 , 王乐禹 . miRs在力学拉伸促进成肌细胞增殖过程中的作用[J]. 中国临床解剖学杂志, 2014 , 32(3) : 306 -311 . DOI: 10.13418/j.issn.1001-165x.2014
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.
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