快骨骼肌亚型肌钙蛋白Ⅰ对鸡前脂肪细胞增殖的影响  

Effects of fast skeletal muscle troponinⅠon proliferation of chicken preadipocytes

作　　者：孔令喆 李金炜 张新宇 旦增晋美 孙泽 孙婴宁[1] KONG Lingzhe;LI Jinwei;ZHANG Xinyu;DANZENG Jinmei;SUN Ze;SUN Yingning(School of Life Sciences,Agriculture and Forestry,Qiqihar University,Qiqihar 161006,China)

机构地区：[1]齐齐哈尔大学生命科学与农林学院,黑龙江齐齐哈尔161006

出　　处：《高师理科学刊》2022年第10期48-53,共6页Journal of Science of Teachers＇College and University

基　　金：国家自然科学基金项目(31402061);2022年度黑龙江省省属高等学校基本科研业务费科研项目(145109211);齐齐哈尔大学创新创业训练计划项目(202220232304)。

摘　　要：实验室前期研究发现,KLF7促进鸡前脂肪细胞增殖,且调控TNNI2基因的表达,但是TNNI2在鸡前脂肪细胞增殖过程中的作用还不清楚.为此,探讨TNNI2基因对鸡前脂肪细胞增殖的影响.qRT-PCR检测永生化鸡前脂肪细胞系(immortalized chicken preadipocyte cell line,ICP1)增殖过程中TNNI2基因的表达情况;将TNNI2过表达载体pCMV-HA-TNNI2转染至ICP1细胞,利用Westernblot实验检测过表达效果.采用CCK-8,EdU法检测过表达TNNI2对细胞增殖的影响,qRT-PCR检测增殖标志基因的表达.TNNI2基因在ICP1细胞增殖过程中呈先下降后逐渐升高的趋势;Westernblot实验结果表明,TNNI2基因过表达成功;CCK-8结果表明,过表达TNNI2后ICP1细胞在450 nm处的吸光值显著上升(P<0.05);EdU结果显示,过表达TNNI2后实验组EdU掺入的细胞数占细胞总数的百分比显著高于对照组(P<0.01);qRT-PCR结果显示,增殖标志基因表达水平有上调趋势,但是差异并不显著(P>0.05).TNNI2基因对鸡前脂肪细胞的增殖具有促进作用.Our previous studies showed that KLF7 promotes the proliferation of chicken preadipocytes and regulates the expression of TNNI2 gene,but the role of TNNI2 in proliferation of preadipocytes remains unclear.Hence,the present study investigated the effect of TNNI2 on the proliferation of chicken preadipocytes.The expression of TNNI2gene during the proliferation of immortalized chicken preadipocytes(ICP1)was detected by qRT-PCR.TNNI2overexpression vector PCMV-HA-TNNI2 was transfected into ICP1 cells.Western blot assay was used to detect the overexpression effect.Cell proliferation was detected by CCK-8 and EdU methods.The expression of proliferation marker gene was detected by qRT-PCR.The TNNI2 gene decreased first and then increased gradually during ICP1cells proliferation.Western blot results showed that TNNI2 gene was overexpressed successfully.CCK-8 results showed that the absorbance value of ICP1 cells at 450 nm increased significantly after TNNI2 overexpression(P<0.05).EdU results showed that after TNNI2 overexpression,the percentage of EdU incorporation in the total number of cells in the experimental group was significantly higher than that in the control group(P<0.01).qRT-PCR results showed that the expression levels of proliferative marker genes were up-regulated but the difference was not significant(P>0.05).TNNI2 gene can promote the proliferation of chicken preadipocytes.

关 键 词：鸡 前脂肪细胞 过表达 TNNI2 增殖 

分 类 号：Q955[生物学—动物学]