目的：探讨恶性黑色素瘤（malignant melanoma，MM）微环境分型对MM患者预后的评估价值。方法：对2010年7月 至2017年5月在南京鼓楼医院手术切除的87例原发性MM组织进行二代测序，免疫组化法检测PD-1、PD-L1、CD3+TIL、MSH2、 MSH6、PMS2和MLH1的表达。随访患者的生存时间，分析不同免疫微环境分型对患者预后的影响及其基因表达特征。结果： 根据PD-L1和TIL表达水平将87例MM患者的肿瘤微环境分为4个亚型：PD-L1+TIL+ 型或双阳型（15/87，17.24%）、PD-L1+TIL型 （15/87，17.24%）、PD-L1- TIL+ 型（20/87，22.99%）、PD-L1- TIL型或双阴型（37/87，42.53%）。双阳型患者的中位无病生存期显著长 于双阴型患者(P<0.05)，此可能与双阴型患者存在更多CDK4、MCL1、MYC、AKT2、CCND1、FGF19等预后不良基因拷贝数扩增 相关；双阳型患者PD-1表达显著高于双阴型患者(P<0.01)，可能与PD-L1、TIL分别与PD-1呈共表达和共不表达有关。结论：根 据PD-L1及TIL表达将MM 患者微环境分为4种亚型，能够区分MM患者预后，双阴型患者存在更多预后不良基因拷贝数扩增。
Objective: To investigate the prognostic value of microenvironmental typing in patients with malignant melanoma (MM). Methods: Next generation sequencing (NGS) was performed on 87 cases of primary MM tissues that had been surgically removed in Nanjing Drum Tower Hospital from July 2010 to May 2017. Immunohistochemistry was carried out to detect the expressions of PD-1, PD-L1, CD3+TIL, MSH2, MSH6, PMS2 and MLH1. The survival of these patients was followed up. Then, the effects of different immune microenvironment types on prognosis and gene expression profile of patients were analyzed. Results: According to the expression level of PD-L1 and TIL, MM patients were divided into 4 immune microenvironment subtypes, including PD-L1+TIL+ group (15/87, 17.24%), PD-L1+ TIL- group (15/87, 17.24%), PD-L1- TIL+ group (20/87, 22.99%), and PD-L1- TIL- group (37/87, 42.53%). The median disease free survival (DFS) time of PD-L1+ TIL+ patients was significantly longer than that of PD-L1- TIL- patients (P<0.05)， which might be related with more copy number amplification of poor prognosis-associated genes in PD-L1- TIL- patients, such as CDK4, MCL1, MYC, AKT2, CCND1 and FGF19. It was found that the expression of PD-1 in PD-L1+ TIL+ patients was significantly higher than that in PD-L1- TIL- ones (P<0.01)，which may be related to the co-expression or un-expression of PD-L1 and PD-1 as well as TIL and PD-1. Conclusion: The microenvironment of MM patients can be divided into four subtypes according to the expression of PD-L1 and TIL, which can distinguish the prognosis of MM patients. The PD-L1- TIL- patients have more copy number amplification of poor prognosis related genes.
国家自然科学基金资助项目（No. 81872484，No. 82073365）；江苏省社会发展面上项目（No. BE2019605）