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目的:观察分析β-catenin基因在人骨肉瘤MG63细胞系的表达变化。方法:培养MG63细胞株,用不同浓度的Wnt3a蛋白或相同浓度、不同时间段的Wnt3a蛋白刺激液MG63细胞生长,用FQ-PCR在m RNA水平检测β-catenin在MG63细胞系的表达变化。培养MG63细胞株,用相同浓度的Wnt3a蛋白刺激,比较刺激组与空白对照组的细胞数目的差别。结果:在细胞生长的第0,1天两组的MG63细胞数量对比没有统计学差异(P>0.05);而在第2,3,4天对照组细胞增殖速度较刺激组缓缓,经比较(t=4.109,3.892,5.215,均P<0.05)。β-catenin基因表达不会因为Wnt3a的浓度增加而增加。100 ng/m L的Wnt3a蛋白刺激β-catenin基因表达最高,其他几个浓度的β-catenin基因表达对比没有统计学差异(P>0.05)。在100 ng/m L的Wnt3a蛋白刺激下,β-catenin基因表达会随时间延长而增加,6 h时β-catenin基因表达最高(P<0.05),随后β-catenin基因表达则没有统计学差异(P>0.005)。结论:Wnt蛋白对MG63细胞有正向增加增殖作用。激活Wnt/β-catenin信号通路能促进MG63的细胞增殖。
OBJECTIVE: To observe and analyze the expression of β-catenin gene in human osteosarcoma MG63 cell line. Methods: The MG63 cell line was cultured and the growth of MG63 cells was stimulated with different concentrations of Wnt3a protein or Wnt3a protein of the same concentration and different time. The expression of β-catenin in MG63 cell line was detected by FQ-PCR at m RNA level. The MG63 cell line was cultured and stimulated with the same concentration of Wnt3a protein to compare the number of cells in the stimulation group and the blank control group. Results: There was no significant difference in the number of MG63 cells between the two groups on day 0 and 1 (P> 0.05). On the 2nd, the 3rd and the 4th day, the proliferation rate of the control group was slower than that of the stimulation group (t = 4.109,3.892,5.215, all P <0.05). β-catenin gene expression will not increase due to the increased concentration of Wnt3a. The expression of β-catenin gene was the highest in Wnt3a protein of 100 ng / m L, while there was no significant difference in the expression of β-catenin in other concentrations (P> 0.05). The β-catenin gene expression increased with time under the stimulation of 100 ng / mL Wnt3a protein, and the expression of β-catenin gene was the highest at 6 h (P <0.05). The expression of β-catenin gene was not significantly different (P> 0.005). Conclusion: Wnt protein has a positive effect on proliferation of MG63 cells. Activation of Wnt / β-catenin signaling promotes cell proliferation in MG63 cells.