基于单细胞测序探索METTL1与胃癌恶性进展的关联性研究

Study on the association between METTL1 and malignant progression of gastric carcinoma based on single-cell sequencing

目的利用单细胞测序分析胃癌免疫治疗关联性基因,并体外实验验证其与胃癌细胞恶性进展关联性。方法使用单细胞测序和GESA寻找差异基因,通过相关预后基因构建随机森林模型,通过预测分析对该模型进行了验证,IHC检测METTL1在胃癌患者中的表达,RT-PCR检测沉默METTL1后的效率,CCK-8、细胞克隆和EDU实验检测沉默METTL1对胃癌细胞增殖效率的影响。结果TCGA数据库和IHC结果均显示METTL1在胃癌组织中高表达。METTL1高表达的患者比METTL1低表达的患者生存率更高(P=0.042)。METTL1及其相关预后基因构建的随机森林模型具有较好的预测价值(AUC=0.863)。CCK-8、细胞克隆和EDU实验显示沉默METTL1抑制了胃癌细胞的增殖。结论单细胞测序筛选的基因所构建的随机森林模型对预测胃癌患者的生存和免疫治疗敏感性具有较高的预测价值,且METTL1参与胃癌细胞恶性进展过程。

Objective To analyze the immunotherapy-associated genes of gastric carcinoma(GC)by single-cell sequencing,and to verify their correlation with malignant progression of GC cells in vitro.Methods Single-cell sequencing and GESA were used to explore the differential genes,random forest model was constructed by related prognostic genes,and then the model was validated by predictive analysis.The expression of METTL1 was detected with IHC in patients with GC,the efficiency was detected with RT-PCR after silencing METTL1,and CCK-8,cell clone,and EDU experiments were used to detect the effect of silencing METTL1 on the proliferation efficiency of GC cells.Results Both database and IHC results showed that METTL1 was highly expressed in GC tissues.Patients with high METTL1 expression had a higher survival rate than those with low METTL1 expression(P=0.042).The random forest model constructed by METTL1 and its related prognostic genes had good predictive value(AUC=0.863).CCK-8,cell clone,and EDU experiments showed that silencing METTL1 inhibited the proliferation of GC cells.Conclusion The random forest model constructed by the genes screened with single-cell sequencing has high predictive value to predict the survival and immunotherapy sensitivity in patients with GC,and METTL1 is involved in the malignant progression of GC cells.