转录因子SIX2基因对牛骨骼肌卫星细胞增殖的影响*

doi:10.12047/j.cjap.6368.2022.113

中国应用生理学杂志 ›› 2022, Vol. 38 ›› Issue (6): 622-627.doi: 10.12047/j.cjap.6368.2022.113

转录因子SIX2基因对牛骨骼肌卫星细胞增殖的影响^*

崔静轩¹, 龚治安¹, 张文天¹, 刘凯¹, 李铁^1.2, 邵淑丽^1.2, 张伟伟^1.2△

1.齐齐哈尔大学生命科学与农林学院,
2.抗性基因工程与寒地生物多样性保护黑龙江省重点实验室, 黑龙江齐齐哈尔 161006

收稿日期:2022-10-18 修回日期:2022-11-25 出版日期:2022-11-28 发布日期:2023-06-12
通讯作者: ^△Tel: 13803619762; E-mail: zww121@163.com
基金资助:
^* 国家青年科学基金项目(31801148);黑龙江省自然科学基金项目(LH2021C099);黑龙江省教育厅基本业务专项(135109260);齐齐哈尔大学研究生创新科研项目(YJSCX2022024)

Effects of transcription factor SIX2 gene on the proliferation of bovine skeletal muscle satellite cells

CUI Jing-xuan¹, GONG Zhi-an¹, ZHANG Wen-tian¹, LIU Kai¹, LI Tie^1,2, SHAO Shu-li^1,2, ZHANG Wei-wei^1,2△

1. College of Life Science and Agroforestry, Qiqihar University,
2. Heilongjiang Key Laboratory of Resistance Genetic Engineering and Biodiversity Protection in Cold Regions, Qiqihar 161006, China

Received:2022-10-18 Revised:2022-11-25 Online:2022-11-28 Published:2023-06-12

摘要/Abstract

摘要： 目的: 研究SIX2基因对牛骨骼肌卫星细胞增殖的影响。方法: 以牛骨骼肌卫星细胞为实验材料,采用实时定量PCR法检测增殖24 h、48 h、72 h时牛骨骼肌卫星细胞中SIX2基因的表达量。通过同源重组构建SIX2基因过表达载体,将SIX2基因过表达质粒和对照组空质粒转染至牛骨骼肌卫星细胞中,每组3个复孔。分别在转染细胞24 h、48 h、72 h时采用MTT法检测细胞活力;转染细胞48 h时,采用流式细胞术检测细胞周期、实时定量PCR(qRT-PCR)和蛋白质免疫印迹(Western blot)检测细胞增殖标志基因的表达。结果: 随着牛骨骼肌卫星细胞增殖,SIX2 mRNA表达升高;与对照组相比,SIX2基因过表达质粒组的SIX2 mRNA和蛋白表达量分别升高了18和2.6倍(P＜0.01)细胞活力增加(P＜0.01),G1细胞比例降低了24.6%,S期和G2期细胞比例分别升高了20.3%、 4.31%(P＜0.01),Pax7基因mRNA和蛋白的表达量分别升高了15.84和1.22倍,增殖标志性基因PCNA、CCNB1 mRNA和蛋白的表达量分别升高了4.82、2.23和1.55、1.46倍(P＜0.01)。结论: 过表达SIX2基因促进牛骨骼肌卫星细胞增殖。

关键词: 骨骼肌卫星细胞, 牛, SIX2, 增殖, 细胞培养

Abstract: Objective: To investigate the effect of SIX2 gene on the proliferation of bovine skeletal muscle satellite cells. Methods: Bovine skeletal muscle satellite cells were used as experimental materials, and the expression of SIX2 gene in bovine skeletal muscle satellite cells was detected by real-time quantitative PCR at 24 h, 48 h, and 72 h of proliferation. The SIX2 gene overexpression vector was constructed by homologous recombination. The SIX2 gene overexpression plasmid and the control empty plasmid were transfected into bovine skeletal muscle satellite cells, and each group had three complex Wells. The cell viability was detected by MTT assay at 24 h, 48 h and 72 h after transfection. At 48 h after transfection, the cell cycle was detected by flow cytometry, and the expressions of cell proliferation marker genes were detected by real-time quantitative PCR (qRT-PCR) and Western blot. Results: With the proliferation of bovine skeletal muscle satellite cells, the expression of SIX2 mRNA was increased. Compared with the control group, the expressions of SIX2 mRNA and protein in the SIX2 gene overexpression plasmid group were increased by 18 and 2.6 times, respectively (P＜0.01). The cell viability of the SIX2 gene overexpression plasmid group was increased (P＜0.01), the proportion of G1 cells was decreased by 24.6%, and the proportion of S phase and G2 phase cells was increased by 20.3% and 4.31%, respectively (P＜0.01). The mRNA and protein expressions of Pax7 gene were increased by 15.84 and 1.22 times, respectively, and the mRNA and protein expressions of proliferation marker genes PCNA and CCNB1 were increased by 4.82, 2.23,1.55 and 1.46 times, respectively (P＜0.01). Conclusion: Overexpression of SIX2 gene promotes the proliferation of bovine skeletal muscle satellite cells.

Key words: skeletal muscle satellite cells, bovine, SIX2, proliferation, cell culture

中图分类号:

Q291

崔静轩, 龚治安, 张文天, 刘凯, 李铁, 邵淑丽, 张伟伟. 转录因子SIX2基因对牛骨骼肌卫星细胞增殖的影响^*[J]. 中国应用生理学杂志, 2022, 38(6): 622-627.

CUI Jing-xuan, GONG Zhi-an, ZHANG Wen-tian, LIU Kai, LI Tie, SHAO Shu-li, ZHANG Wei-wei. Effects of transcription factor SIX2 gene on the proliferation of bovine skeletal muscle satellite cells[J]. CJAP, 2022, 38(6): 622-627.

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转录因子SIX2基因对牛骨骼肌卫星细胞增殖的影响^*

Effects of transcription factor SIX2 gene on the proliferation of bovine skeletal muscle satellite cells

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