GREM1对胃癌细胞增殖迁移的作用

Effect of GREM1 on the proliferation and metastasis of gastric cancer cells

目的检测胃癌细胞中GREM1基因的表达,探讨其对胃癌细胞生物学特性的影响并评估其在胃癌诊断和胃癌患者预后中的临床价值。方法运用数据库分析GREM1基因在胃癌组织和癌旁组织中的表达差异,评估GREM1基因表达水平对胃癌患者预后的相关性。Western blot检测胃癌细胞系中GREM1蛋白质表达水平。AGS细胞中沉默GREM1基因后,采用平板克隆、Transwell和western blot检测其对胃癌细胞增殖、迁移、上皮间质转化(EMT)发生及Wnt/β-catenint通路的影响。结果 Kaplan-Meier分析表明,GREM1基因高表达患者总体生存率(OS)和无进展生存率(PFS)均降低。GREM1蛋白在AGS细胞系中的表达水平(1.967±0.056)最高。平板克隆、Transwell及western blot实验显示,沉默GREM1基因可导致胃癌细胞的增殖及迁移能力降低(t分别为22.00,29.60;P均<0.01);E-cadherin表达上升(t=10.65,P<0.01),ZEB1、MMP2表达均下降(t分别为10.74和13.67,P均<0.01);Wnt/β-catenin通路中β-catenin、CyclinD1、c-myc、p-GSK3β和PCNA的表达水平均降低(t分别为12.65,16.21,8.74,7.75和8.42;P均<0.01)。结论 GREM1通过激活Wnt/β-catenin诱导EMT发生,促进肿瘤转移和生长。GREM1可作为新的胃癌分子诊断和预后指标。

Objective To detect the expression of GREM1 gene in gastric cancer cells, investigate its effects on the biological characteristics of gastric cancer cells and evaluate its application value in the diagnosis and prognosis of gastric cancer. Methods The expression difference of GREM1 in gastric cancer tissues and adjacent normal tissues was analyzed by the database, and the correlation of GREM1 expression levels with the prognosis of gastric cancer patients was evaluated. The expression levels of GREM1 protein in gastric cancer cell lines were detected by western blot. After GREM1 gene in AGS cells was silenced, its effects on the proliferation, migration, epithelial-mesenchymal transition(EMT) and Wnt/β-catenin pathway of AGS cells were detected by the colony formation assay, Transwell and Western blot, respectively. Results Kaplan-Meier analysis showed that the patients with high expression of GREM1 gene had low overall survival(OS) and progression-free survival(PFS). The expression level of GREM1 protein in AGS cells was the highest in all gastric cancer cell lines(1.967 ± 0.056). The analysis of colony formation assay, Transwell and Western blot showed that the silencing of GREM1 gene could decrease the proliferation and migration of gastric cancer cells(t=22.00;t=29.60;P<0.01), increase the expression of E-cadherin(t=10.65, P<0.01), and decrease the expressions of ZEB1 and MMP2(t=10.74;t=13.67;P<0.01) and the expressions of β-catenin, Cyclin D1, c-myc, p-GSK3β and PCNA in the Wnt/β-catenin pathway(t=12.65;t=16.21;t=8.74;t=7.75;t=8.42;P<0.01). Conclusion GREM1 may induce EMT by activating the Wnt/β-catenin pathway, and promote the metastasis and growth of tumors, which may be used as a new molecular diagnostic and prognostic marker for gastric cancer.