[转录因子SIX2基因对牛骨骼肌卫星细胞增殖的影响]。

Jing-Xuan Cui, Zhi-An Gong, Wen-Tian Zhang, Kai Liu, Tie Li, Shu-Li Shao, Wei-Wei Zhang
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引用次数: 0

摘要

目的:探讨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,在增殖24 h、48 h、72 h时,采用实时荧光定量PCR检测SIX2基因在牛骨骼肌卫星细胞中的表达情况。采用同源重组法构建SIX2基因过表达载体。将SIX2基因过表达质粒和对照空质粒转染牛骨骼肌卫星细胞,每组有3个复孔。转染后24 h、48 h和72 h采用MTT法检测细胞活力。转染后48 h,流式细胞术检测细胞周期,实时荧光定量PCR (qRT-PCR)和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的增殖,six2mrna的表达增加。与对照组相比,SIX2基因过表达质粒组SIX2 mRNA和蛋白的表达量分别提高了18倍和2.6倍(P<0.01)。SIX2基因过表达质粒组细胞活力升高(P<0.01), G1期细胞比例降低24.6%,S期和G2期细胞比例分别升高20.3%和4.31% (P<0.01)。Pax7基因mRNA和蛋白表达量分别增加了15.84倍和1.22倍,增殖标志基因PCNA和CCNB1 mRNA和蛋白表达量分别增加了4.82倍、2.23倍、1.55倍和1.46倍(P<0.01)。结论:SIX2基因的过表达促进了牛骨骼肌卫星细胞的增殖。
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[Effects of transcription factor SIX2 gene on the proliferation of bovine skeletal muscle satellite cells].

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.

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