基于单细胞RNA测序的胃癌淋巴转移的细胞学特征及分子机制研究

Cytological characteristics and molecular mechanism of lymphatic metastasis in gastric cancer based on single cell RNA sequencing

目的探讨胃腺癌淋巴转移过程中的细胞学特征及相关分子机制的改变。方法收集2019年9-12月该院胃肠外科13例行胃癌根治术患者的胃癌组织及癌旁组织,利用单细胞RNA测序技术揭示标本中的细胞学特征及相关标记基因。利用实时定量PCR(RT-qPCR)检测主细胞3的标记基因XIST和窝状黏液细胞3标记基因CXCL5的表达水平。分析XIST、CXCL5在淋巴结转移阳性和阴性胃癌患者表达水平的差异。结果主细胞3和窝状黏液细胞3在胃癌淋巴转移阳性的癌组织明显富集。主细胞3的标记基因XIST在淋巴转移阳性患者胃癌组织表达水平为(0.297±0.261),高于淋巴转移阴性患者胃癌组织的表达水平(0.025±0.023),差异有统计学意义(P<0.05)。窝状黏液细胞3的标记基因CXCL5在淋巴转移阳性患者胃癌组织表达水平为(0.036±0.019),高于其在淋巴转移阴性患者胃癌组织的表达水平(0.008±0.007),差异有统计学意义(P<0.05)。拟时序分析发现主细胞可能是胃癌发生的起源。结论XIST、CXCL5可能是胃腺癌淋巴转移的潜在信号。

Objective To investigate the changes in cytological characteristics and related molecular mechanism in the process of the lymphatic metastasis in gastric adenocarcinoma.Methods The gastric cancer tissues and paracancerous tissues from 13 patients undergoing radical gastrectomy in the department of gastrointestinal surgery in this hospital from September to December 2019 were collected.The single-cell RNA sequencing technology was used to detect the cytological characteristics and related marker genes in the samples.Real-time quantitative PCR(RT-qPCR)was used to detect the expression levels of the marker gene XIST in chief cell 3 and the marker gene CXCL5 in pit mucous cell 3.The differences in the expression levels of XIST and CXCL5 in patients with lymph node metastasis and non-metastatic gastric cancer were analyzed.Results The chief cell 3 and pit mucous cell 3 were significantly enriched in the gastric cancer tissues with positive lymphatic metastasis.The expression level of chief cell 3 marker gene XIST was(0.297±0.261)in positive patients with lymphatic metastasis,which was higher than that in negative patients with lymphatic metastasis(0.025±0.023).The difference was statistically significant(P<0.05).The expression level of CXCL5,a marker gene of pit mucous cell 3 was(0.036±0.019)in the gastric cancer tissues of patients with positive lymphatic node metastasis,which was higher than that in the gastric cancer tissues of patients with negative lymphatic node metastasis(0.008±0.007).The difference was statistically significant(P<0.05).The quasi-time sequence analysis showed that the main cell might be the origin of gastric cancer.Conclusion XIST and CXCL5 might be the potential signals of lymphatic metastasis in gastric adenocarcinoma.