RTN4在胃癌发病机制中的作用

The Role of RTN4 in the Pathogenesis of Gastric Carcinoma

目的了解RTN4蛋白在胃癌发病过程中的作用,为揭示胃癌发病的分子机制提供实验资料。方法将pIRESneo3-RTN4真核表达载体转染胃粘膜上皮GES1细胞,建立RTN4高表达的GES1-RTN4细胞系;通过细胞生长曲线、平皿克隆与软琼脂集落形成实验、流式细胞仪,观察RTN4高表达对GES1细胞生长与增殖、周期与凋亡的影响;利用Western-blot检测GES1-RTN4细胞中IκBα蛋白的表达。结果 GES1-RTN4细胞的生长速度较GES1和GES1-pIRESneo3细胞加快,统计学意义显著(P<0.01);GES1-RTN4细胞较GES1和GES1-pIRESneo3细胞克隆体积大、数目多,统计学意义显著(P<0.01);GES1-RTN4细胞中S期细胞比例较GES1和GES1-pIRESneo3细胞增加,细胞增殖指数增加,细胞凋亡率降低,统计学意义显著(P<0.01);GES1-RTN4细胞中IκBα蛋白表达较GES1和GES1-pIRESneo3细胞减少,统计学意义显著(P<0.01)。结论RTN4通过活化NF-κB信号通路,提高细胞增殖指数,抑制细胞凋亡,促进GES1细胞的增殖。

Objective To investigate the role of RTN4 protein in the pathogenesis of gastric carcinoma and to provide experimental data for revealing the molecular mechanism of gastric carcinoma.Methods The plRESneo3-RTN4 eukaryotic expression vector was transfected into gastric mucosal epithelial GES1 cells to establish a high expression of GES1-RTN4 cell line. The cell growth curve, plate clone and soft agar colony formation assay, flow cytometry, high RTN4 expression were observed. The effects of GES1 cell growth and proliferation, cycle and apoptosis were detected by Western-blot. The expression of I KBc~ protein in GES1-RTN4 cells was detected by Western-blot. Results The growth rate of GES1-RTN4 cells was faster than that of GES1 and GES1 -plRESneo3 cells, which was statistically significant （P〈0.01）. The GES1-RTN4 cells were larger and more numerous than GES1 and GES1- plRESneo3 cells,the statistical significance was significant （P〈0.01）; the proportion of S phase cells in GES1-RTN4 cells was higher than that of GES1 and GES1-plRESneo3 cells, the cell proliferation index increased, and the apoptosis rate decreased, statistically significant （P〈0.01）;The expression of IKBc~ protein in GES1 -RTN4 cells was significantly lower than that in GES1 and GES1 - plRESneo3 cells,statistically significant （P〈0.01）. Conclusion RTN4 can increase the cell proliferation index, inhibit cell apoptosis and promote the proliferation of GES1 cells by activating NF-KB signaling pathway.