摘要
[目的]本文旨在探究UCP3基因对C2C12成肌细胞增殖的调控作用。[方法]构建慢病毒包被的UCP3基因过表达和干扰载体,转染C2C12细胞,分别促进和抑制UCP3基因的表达;通过CCK-8和EDU染色试验检测C2C12细胞的增殖效果,采用qRT-PCR技术检测细胞增殖和凋亡的相关基因表达量的变化,利用Western Blot方法检测细胞增殖关键蛋白PCNA的变化。[结果]CCK-8和EDU染色试验结果发现,与过表达对照组(OE-NC)相比,UCP3基因过表达组(OE-UCP3)细胞的增殖速度极显著降低(P<0.01),细胞增殖相关基因PCNA和Ki67的mRNA表达量极显著降低(P<0.01),细胞凋亡相关基因Bax和caspase3的表达量极显著升高(P<0.01);细胞增殖关键蛋白PCNA的含量也极显著降低(P<0.01)。与干扰对照组(Sh-NC)相比,UCP3基因干扰组(Sh-UCP3)细胞的增殖速度显著增加(P<0.05),细胞增殖相关基因PCNA和Ki67的mRNA表达量极显著增加(P<0.01),细胞凋亡相关基因Bax和caspase3的表达量显著降低(P<0.05),细胞增殖关键蛋白PCNA的含量也显著升高(P<0.05)。[结论]UCP3基因可能通过抑制细胞增殖基因PCNA和Ki67的表达,促进细胞凋亡基因Bax和caspase3的表达而抑制C2C12细胞的增殖。该结果为骨骼肌中UCP3的功能研究提供基础数据。
[Objective]The aim of this study was to investigate the regulatory effect of UCP3 gene on the proliferation of C2 C12 myoblasts.[Methods]The lentivirus-coated UCP3 gene overexpression and interference vectors were constructed and transfected into C2 C12 cells to promote or inhibit the expression of UCP3 gene,respectively.The proliferation rate of C2 C12 cells were measured by CCK-8 experiment and EDU staining tests.qRT-PCR technology was used to detect the genes expression variation that related to cell proliferation and apoptosis.Western Blot was used to detect the changes of PCNA,a key protein for cell proliferation.[Results]CCK-8 and EDU staining experiments showed that the proliferation rate of C2 C12 cell in the UCP3 gene overexpression group(OE-UCP3)was significantly decreased significantly compared to the overexpression control group(OE-NC)(P<0.01).The mRNA expression of cell proliferation-related genes(PCNA and Ki67)and apoptosis-related genes(Bax and caspase3)in OE-UCP3 group were significantly decreased or increased respectively compared to the overexpression control group(OE-NC)(P<0.01).The content of PCNA,a key protein for cell proliferation,was also significantly decreased compared to(OE-NC)group(P<0.01).Moreover,Compared to the interference control group(Sh-NC),the proliferation rate of C2 C12 cell in UCP3 gene interference group(Sh-UCP3)was enhanced increased significantly(P<0.05)compared to the interference control group(Sh-NC),and the mRNA expression of cell proliferation-related genes(PCNA and Ki67)and apoptosis-related genes(Bax and caspase3)were either increased or decreased(P<0.01,P<0.05),respectively.The content of cell proliferation key protein PCNA was also increased significantly in Sh-UCP3 group(P<0.05).[Conclusion]UCP3 gene may inhibit the proliferation of C2 C12 cells by down-regulating the expression of cell proliferation-related genes and up-regulating the expression of apoptosis-related genes.
作者
何志强
黑伟
张万锋
张燕伟
吴怡琦
蔡春波
杨阳
高鹏飞
李步高
郭晓红
He Zhiqiang;Hei Wei;Zhang Wanfeng;Zhang Yanwei;Wu Yiqi;Cai Chunbo;Yang Yang;Gao Pengfei;Li Bugao;Guo Xiaohong(College of Animal Science,Shanxi Agricultural University,Taigu 030801)
出处
《山西农业大学学报（自然科学版）》
CAS
北大核心
2021年第2期74-82,共9页
Journal of Shanxi Agricultural University(Natural Science Edition)
基金
国家自然科学基金(31872336)
三晋学者支持计划专项经费(2016
2017)
山西省青年基金(201901D211376,201901D211369)。
