COL3A1基因的一种新转录本对固原鸡成肌细胞增殖的影响  

Effects of a novel transcript of COL3A1 gene on the proliferation of Guyuan chicken myoblasts

作　　者：蔡翠翠 万鹏 杨雪瑶 雷初朝[2] 陈宏[2] 李玉莲 CAI Cuicui;WAN Peng;YANG Xueyao;LEI Chuchao;CHEN Hong;LI Yulian(Guyuan Branch,Ningxia Academy of Agricultural and Forestry Sciences,Guyuan 756000,China;College of Animal Science and Technology,Northwest Agriculture and Forestry University,Yangling 712100,China)

机构地区：[1]宁夏农林科学院固原分院,宁夏固原756000 [2]西北农林科技大学动物科技学院,陕西杨凌712100

出　　处：《黑龙江畜牧兽医》2021年第10期34-40,154,共8页Heilongjiang Animal Science And veterinary Medicine

基　　金：宁夏回族自治区农业科技自主创新资金项目(DWHZC-2017009,DW-X-2020010)。

摘　　要：为了研究鸡Ⅲ型胶原蛋白α1链(collagen type Ⅲ alpha 1 chain,COL3A1)基因的可变剪接状况及新转录本功能,试验以固原鸡15天鸡胚(胚胎期)和180日龄成年鸡(成年期)的心脏、肝脏、脑、肺脏、肾脏、肌肉组织为样品,从15天鸡胚组织样品中分离与培养原代成肌细胞,采用实时荧光定量PCR技术、CCK-8试验、EdU试验、流式细胞技术等鉴定新转录本,分析其表达量,并研究其对固原鸡成肌细胞增殖的影响。结果表明:发现并鉴定出固原鸡COL3A1基因的一种新转录本,命名为Ⅲ型胶原蛋白α1链-a(collagen type Ⅲ alpha 1 chain-1,COL3A1-a);COL3A1-a在胚胎期肺脏和成年期肌肉组织中表达量较高;干扰固原鸡成肌细胞COL3A1-a的表达,使得COL3A1-a的表达量约下降44%(P<0.05),导致促增殖标志基因PCNA的表达量显著降低(P<0.05),抑增殖标志基因p21的表达量显著升高(P<0.05);干扰COL3A1-a的表达24 h后,细胞活力约下降了10%(P<0.05),EdU阳性细胞数量明显减少,EdU阳性细胞比例约降低了10%(P<0.01),S期细胞比例减少,成肌细胞生长受到抑制。说明干扰COL3A1-a能够抑制固原鸡成肌细胞的增殖,即COL3A1能够促进固原鸡成肌细胞的增殖,研究COL3A1-a有助于解析固原鸡优异生长性状的遗传机制。In order to study the alternative splicing status and function of new transcripts of the chicken collagen typeⅢalpha 1 chain(COL3 A1)gene,15-day Guyuan embryo(embryonic stage)and tissues(heart,liver,brain,lung,kidney,and muscle)of 180-day adult(adult stage)chickens were used as experimental samples.Primary myoblasts were isolated and cultured from 15-day chicken embryo tissue samples.Real-time fluorescent quantitative PCR technology,CCK 8 trials,EdU,and flow cytometry were used to identify the new transcript and analyze its expression,so as to study its effect on the proliferation of Guyuan chicken myoblasts.The results showed that a new kind of chicken COL3 A1 gene transcript was found and identified,it was named the chicken collagen typeⅢalpha 1 chain-1(COL3 A1-a).Its expression level was high in embryonic lung and adult muscle tissue.After the expression of COL3 A1-A in Guyuan chicken myoblasts was disrupted,the expression of COL3 A1-a was decreased by 44%(P<0.05),which resulted in a significant decrease in the expression of pro-proliferation marker gene PCNA(P<0.05),and a significant increase in the expression of anti-proliferation marker gene p21(P<0.05).After interfering with the expression of COL3 A1-A for 24 h,the cell viability decreased by 10%(P<0.05),the proportion of EdU positive cells was significantly reduced,and the proportion of EdU positive cells was decreased by 10%(P<0.01).The proportion of S phase cells decreased and the growth of myoblasts was inhibited.The results indicated that interfering with COL3 A1-A could inhibit the proliferation of Guyuan chicken myoblasts,that was,COL3 A1 could promote the proliferation of Guyuan chicken myoblasts.The study of COL3 A1-A in Guyuan chickens was helpful to elucidate the genetic mechanism of excellent growth traits in Guyuan chickens.

关 键 词：固原鸡 转录本 COL3A1 肌肉 增殖 

分 类 号：S813.3[农业科学—畜牧学]