不同TERT基因状态的肝细胞癌患者的临床特征及外泌体mRNA表达谱探索

The clinical characteristics and exosomal mRNA expression profile of hepatocellular carcinoma patients with different TERT gene status

目的分析不同端粒酶逆转录酶(TERT)基因状态的肝细胞癌患者的临床特征及外泌体mRNA表达谱,探索不同TERT基因状态在原发性肝癌(简称肝癌)发生、发展过程中的潜在作用差异。方法纳入36例行肝切除术的初治肝癌患者,对患者术前外周血进行外泌体mRNA测序分析,检测手术切除的肿瘤组织TERT基因突变及其与乙型肝炎病毒(HBV)整合情况,并根据TERT基因状态将患者分为TRET启动子区(TERTp)突变组、HBV-TERTp整合组、无TERT基因整合及突变组,比较3种TERT基因状态的临床特征、实验室检查及外泌体mRNA表达特征。结果TERTp整合与TERTp突变互斥,36例患者中,HBV-TERTp整合组9例(25.00%),TERTp突变组13例(36.11%),无TERT基因整合及突变组14例(38.89%);HBV-TERTp整合组与无TERT基因整合及突变组的临床特征及实验室结果相似,将这两组合并为非TERTp突变组,该组与TERTp突变组相比,TERTp突变组患者年龄更大(P=0.036),甲胎蛋白(AFP)水平和阳性率更低(P=0.044、0.017),中性粒细胞/淋巴细胞比值(NLR)更低(P=0.025),肿瘤最大径、维生素K缺乏或拮抗剂Ⅱ诱导的蛋白(PIVKA-Ⅱ)、血小板(PLT)、纤维蛋白原(Fib)水平更低。外泌体差异mRNA富集分析显示,TERTp突变组与非TERTp突变组间的差异基因在线粒体相关的细胞组分、分子功能及生物过程明显富集。结论TERTp突变组与非TERTp突变组患者的临床特征存在显著差异,TERTp突变组患者年龄更大,具有更温和的实验室特征;用AFP作为肿瘤筛检指标,可能会造成漏检;TERTp突变可能通过增强线粒体代谢等方式获能以促进肿瘤增殖,抗氧化治疗可能是TERTp突变肝癌患者有效的治疗途径。

Objective To analyze the clinical characteristics and exosomal mRNA expression profiles of hepatocellular carcinoma patients with different telomerase reverse transcriptase(TERT)gene status,and to explore the potential differences of TERT gene status in the development of primary liver cancer(referred to as liver cancer).Methods Thirty-six patients with primary liver cancer who underwent hepatectomy were enrolled.The peripheral blood of the patients was subjected to exosomal mRNA sequencing analysis to detect the mutation of TERT gene in surgically resected tumor tissue and the integration of hepatitis B virus(HBV).According to the TERT gene status,the patients were divided into TERT promoter region(TERTp)mutation group,HBV-TERTp integration group,no TERT gene integration and mutation group,and the clinical characteristics,laboratory examination and exosomal mRNA expression characteristics of the three TERT gene states were compared.Results TERTp integration and TERTp mutation were mutually exclusive.Among 36 patients,9 patients(25.00%)were in the HBV-TERTp integration group,13 patients(36.11%)were in the TERTp mutation group,and 14 patients(38.89%)were in the non-TERT gene integration and mutation group.The clinical characteristics and laboratory results of the HBV-TERTp integration group and the non-TERT gene integration and mutation group were similar.The two groups were combined into the non-TERTp mutation group.Compared with the TERTp mutation group,the patients in the TERTp mutation group were older(P=0.036),alpha-fetoprotein(AFP)level and positive rate were lower(P=0.044,0.017),neutrophil/lymphocyte ratio(NLR)was lower(P=0.025),maximum diameter of tumor,vitamin K deficiency or antagonistⅡ-induced protein(PIVKA-Ⅱ),platelets(PLT),and fibrinogen(Fib)levels were lower.Exosomal differential mRNA enrichment analysis showed that the differential genes between TERTp mutant group and non-TERTp mutant group were significantly enriched in mitochondrial-related cellular components,molecular functions and biological processes.Conclusion There are significant differences in clinical characteristics between TERTp mutation group and non-TERTp mutation group,patients in TERTp mutation group are older and have milder laboratory characteristics.Using AFP as a tumor screening index may cause missed detection.TERTp mutation may enhance tumor proliferation by enhancing mitochondrial metabolism,antioxidant therapy may be an effective treatment approach for patients with TERTp mutation liver cancer.