干扰E2F7对C2C12成肌细胞增殖的影响  

Effect of Interfering with E2F7 on the Proliferation of C2C12 Myoblasts

作　　者：范源[1] 甘麦邻 罗嘉[1] 谭娅[1,2] 张顺华 朱砺[1] FAN Yuan;GAN Mailin;LUO Jia;TAN Ya;ZHANG Shunhua;ZHU Li(College of Animal Science and Technology, Sichuan Agricultural University, Chengdu 611130, China;Institute of Animal Husbandry and Veterinary, Guizhou Academy of Agricultural Science, Guiyang 550005, China)

机构地区：[1]四川农业大学动物科技学院,四川成都611130 [2]贵州省农业科学院畜牧兽医研究所,贵州贵阳550005

出　　处：《云南农业大学学报（自然科学版）》2019年第3期408-413,共6页Journal of Yunnan Agricultural University:Natural Science

基　　金：国家重点研发计划项目(2018YFD0501004);农业农村部农业重大技术协同推广计划试点项目;四川省科技支撑计划项目(2017NFP0135;2016NZ0089;2016NYZ0050)

摘　　要：【目的】探究转录因子E2F7在骨骼肌细胞发育过程中的作用。【方法】以C2C12成肌细胞为研究对象,通过设计小干扰RNA干扰C2C12成肌细胞中E2F7的表达,分为干扰组和对照组,并利用RT-PCR、CCK-8和EdU等技术检测在C2C12成肌细胞中干扰E2F7后对成肌细胞增殖的影响。【结果】E2F7 mRNA表达水平在C2C12成肌细胞增殖期极显著高于分化期(P<0.01);与对照组相比,干扰组的细胞活率和新生细胞比率皆极显著高于对照组(P<0.01);干扰E2F7显著提升了C2C12成肌细胞中Cyclin E的表达水平(P<0.05),极显著促进Cyclin D和CDK4的表达(P<0.01);并且干扰E2F7可显著促进E2F2 (P<0.05)和极显著促进E2F1及E2F3的表达(P<0.01),同时极显著促进与成肌细胞增殖相关microRNAs (miR-7、miR-25、miR-27和miR-92a)的表达(P<0.01)。【结论】干扰E2F7可促进C2C12成肌细胞的增殖,E2F7可能是通过调控经典E2Fs信号通路和成肌细胞增殖相关microRNA发挥作用。[Purpose]To explore the biological role of E2F7 in the development of skeletal muscle cells.[Method]C2C12 myoblasts were used as the research object. Small interfering RNA was designed to interfere with the expression of E2F7 in C2C12 myoblast. C2C12 myoblasts were divided into interfering group and control group. The effect of E2F7 on the proliferation of C2C12 myoblasts was detected by RT-PCR, CCK-8 assay and EdU assay, respectively.[Result]The expression of E2F7 was significantly higher in C2C12 myoblasts during the proliferative phase than that in the differentiation phase (P<0.01);the viability and ratio of neoplastic cells of C2C12 myoblasts after E2F7 interference were extremely significantly higher than those of the control group (P<0.01). The expression level of Cyclin E was significantly increased (P<0.05), and the expression of Cyclin D and CDK4 was extremely significantly elevated when C2C12 myoblasts interfered with E2F7 (P<0.01). Further finding showed that interference with E2F7 significantly promoted E2F2 (P<0.05) and extremely significantly promoted the expression of E2F1 and E2F3 (P<0.01). At the same time, it extremely significantly promoted the expression levels of microRNAs (miR-7, miR-25, miR-27, and miR-92a) which associated with myoblasts proliferation (P<0.01).[ Conclusion] The results suggest that interference with can promote the proliferation of C2C12 myoblasts, and E2F7 may plays a role through regulating the classical E2Fs signaling pathway and myoblasts proliferation-related microRNAs.

关 键 词：E2F7 C2C12成肌细胞 增殖 小干扰RNA 骨骼肌发育 

分 类 号：Q344.5[生物学—遗传学]