基于二代测序技术探究胃癌中医证型的基因突变特征

Gene Mutations of Gastric Cancer with Different Traditional Chinese Medicine Syndromes Detected with Next Generation Sequencing

[目的]利用二代测序技术研究胃癌中医证型的基因突变特征,从肿瘤基因组层面初步探索胃癌中医证候的客观化指标,指导临床精准治疗。[方法]收集临床病理分期为Ⅰ~Ⅳ期且未经任何治疗的胃癌患者,根据2011年《胃癌中医诊疗方案》进行辨证分型,采用二代测序技术(含450个肿瘤相关突变基因)检测肿瘤细胞的基因突变情况,结合临床资料和中医证型进行生物信息学分析,从而探究胃癌中医证型的基因突变特点。[结果]共有130例Ⅰ~Ⅳ期胃癌患者纳入本研究,辨证分型为脾气虚证、血虚证、热毒证等共8个单证。对中医单证和测序获得的363个突变基因进行分析,得到与各单证相关的高频和驱动基因,脾气虚证多见ARID1A、PIK3CA、APC基因突变;血虚证多见KMT2C基因突变;热毒证多见TGFBR2、HNF1A、ERBB3、KMT2D基因突变。130例患者中共有92例携带与单证证型相关的突变基因,采用无监督层次聚类分析得到脾虚热毒证这一特征证型,该复合证型多有Hippo、TGF-β等信号通路的激活,表现为微卫星高度不稳定(MSI-H)和高肿瘤突变负荷(TMB)的特点。[结论]不同单证的胃癌患者具有不同的基因突变特征,脾气虚证多有PIK3CA、ARID1A、APC基因突变;新发胃癌患者多为虚实夹杂的复合证型,辨证为脾虚热毒证的胃癌患者可能是免疫治疗的适宜人群。

[Purpose]To explore the gene mutation patterns of gastric cancer with different traditional Chinese medicine(TCM)syndromes detected with next generation sequencing(NGS).[Methods]One hundred and thirty patients with TNM stageⅠ~Ⅳgastric cancer were enrolled in the study.The Gastric Cancer TCM Diagnosis and Treatment Guidelines published in 2011 was used for syndrome differentiation.NGS(including 450 tumor-related genes)and bioinformatics methods were used to explore the characteristics of gene mutations in gastric cancer with different TCM syndromes.[Results]The gastric cancer patients were classified as 8 single TCM syndromes.Total363 mutated genes were obtained from sequencing,and 92 patients carried mutated genes related to the single syndrome.The mutations of ARID1 A,PIK3 CA and APC were mainly found in the"Piqixu"syndrome;KMT2 C gene mutation was common in"Xuexu"syndrome;TGFBR2,HNF1 A,ERBB3 and KMT2 D gene mutations were commonly found in"Redu"syndrome.The unsupervised hierarchical cluster analysis was used to obtain the characteristic syndrome type of"Piqixu"syndrome with"Redu"syndrome.Most of the complex syndrome activated the Hippo and TGF-βsignal pathway,which were characterized by microsatellite instability-high(MSI-H)and high tumor mutation burden(TMB).[Conclusion]Gastric cancer patients with different single TCM syndrome have characteristic gene mutations,and most of them were PIK3 CA,ARID1 A,APC gene mutations.Patients with newly diagnosed gastric cancer are likely to have complex TCM syndromes mixed with deficiency and excess.Patients with the complex syndromes of"Pixu with Redu"may be suitable for immunotherapy.