44例头颈部鳞状细胞癌多原发癌患者M2型巨噬细胞浸润特点及其临床意义分析

Analysis of M2 macrophage infiltration and its clinical significance in 44 patients with multiple primary cancers of the head and neck

目的探讨分析M2型巨噬细胞在头颈部鳞状细胞癌(squamous cell carcinoma of head and neck,HNSCC)多原发癌患者中的浸润特点及其临床意义,总结及发掘M2型巨噬细胞在多原发癌患者鉴别诊断及预后评价中的作用。方法获取癌症基因组图谱(The Cancer Genome Atlas,TCGA)数据库中502例HNSCC患者的癌组织与44例癌旁组织样本RNA表达谱数据,使用R包及R软件v4.0.3进行统计分析。回顾性筛选1998年7月至2016年2月期间就诊于北京大学口腔医学院·口腔医院口腔颌面外科,经随访并确诊为HNSCC多原发癌的患者44例,其中男性17例,女性27例。筛选2013年8月至2015年12月期间经随访为单发的牙龈癌患者41例,其中男性28例,女性13例。石蜡切片免疫组织化学染色全景切片分析CD163表达阳性细胞数及表达特点,CD163阳性细胞数≤15为CD163低表达组,CD163阳性细胞数>15为CD163高表达。采用χ2检验及Spearman相关性分析比较单发牙龈癌患者与HNSCC多原发癌患者CD163阳性计数和(或)浸润深度与发病次数间差异及关联。多原发癌及单发牙龈癌患者的临床特征用SPSS 25.0进行描述性统计分析。结果分析TCGA数据库RNAseq数据结果提示,巨噬细胞浸润在HNSCC组织中较癌旁组织增加(P<0.001)。多原发癌患者原发肿瘤的CD163阳性细胞数[90.9%(40/44)]显著高于单发牙龈癌患者[61.0%(25/41)](P=0.001),且CD163阳性细胞计数与发病次数呈正相关(r=0.368,P=0.001)。CD163原发肿瘤阳性细胞数与浸润深度比值与发病次数也呈正相关(r=0.331,P=0.03)。44例多原发癌患者以女性、无烟酒嗜好、无系统病史、Tis~T2期以及N0期HNSCC为主,多原发次数自第2至第8次不等。同时癌发病比例随多原发次数增加而增大。原发癌发病部位以舌、牙龈、颊为主,随着多原发次数增加,发病部位牙龈、颊及腭部占比增高。结论M2型巨噬细胞数和(或)结合浸润深度与多原发癌发生次数相关,可能成为临床鉴别单发与多原发癌患者的指标。对发生于头颈部(舌、牙龈、颊部为主)的女性、无烟酒嗜好、无系统病史的早期HNSCC患者,应加强随访。

Objective To investigate and analyze the characteristics of M2 macrophage infiltration and the clinical significance in patients with multiple primary cancers(MPCs)of head and neck in order to explore its role in the diagnosis and prognosis for patients with MPCs.Methods RNA-seq data were downloaded from the Genomic Data Commons data portal(TCGA)and the R software v4.0.3 was used to statistically analyze the differences.A retrospective analysis was conducted by screening the clinical data of 44 patients(17 males and 27 females)with MPCs in head and neck from July 1998 to February 2016 in the Department of Oral and Maxillofacial Surgery,Peking University School and Hospital of Stomatology.Clinical data from a batch of 41 patients(28 males and 13 females)with gingival cancer and without MPCs from August 2013 to December 2015 were collected and analyzed.The number of CD163 positive cells and the expression patterns in immunohistochemically panoramic slices were observed under high magnification.Chi-square test and Spearman correlation analysis were used to compare the difference and correlation between the CD163 positive counts and/or depths of invasion and the number of incidences.The descriptive statistics on the clinical features was performed by SPSS 25.0.Results TCGA database analysis showed that the infiltration of macrophage in patients with squamous cell carcinoma of head and neck(HNSCC)was increased compared to the para-cancer sites.A total of 142 tissue samples from 44 patients with MPCs were selected in the present single-center retrospective research.The number of CD163-positive cells in MPCs patients[90.9%(40/44)]was significantly increased compared to single gingival cancer patients[61.0%(25/41)](r=0.353,P=0.001),which was related to the number of occurrence(r=0.368,P=0.001).The ratio of the CD163 counts in primary tumor to the depths of invasion was positively correlated with the number of onsets(r=0.331,P=0.03).In terms of clinical features,the 44 patients with MPCs were mainly female,non-smoking,no alcohol addiction,no systemic history,Tis-T2 stage and N0 stage squamous cell carcinoma.The number of incidences ranged from two to eight.The incidence of cancer relative to synchronous cancer increased with the increased occurrence of MPCs.The primary cancer mainly occurred in tongue,gingiva and buccal sites,while the proportion of onset sites in gingiva,buccal and palate areas increased with the increased occurrence.Conclusions M2 type macrophage counts and/or ratio to depth of infiltration were associated with the occurrence of MPCs,which could be used as a clinical indicator to distinguish single and MPCs in HNSCC.For early stage of HNSCC,patients with clinical characters of women,non-smoking,no alcohol addiction,no systemic medical history and sites of tongue,gingiva,and buccal should be paid more attention on their follow-up plan.The findings in the present study was also helpful to explore new treatment methods for the patients with MPCs.