原发性肝癌患者肿瘤微环境中CD8+细胞毒性T淋巴细胞浸润水平与预后的关系探究

摘要/Abstract

摘要： 目的探讨原发性肝癌(PLC)患者肿瘤微环境中CD8⁺细胞毒性T淋巴细胞的浸润程度与患者预后之间的关系,以期为肝癌的治疗提供新的免疫学依据。方法本研究为回顾性研究,选取2018年1月至2020年1月间进行PLC切除术治疗的患者68例为研究对象。患者行切除术时取新鲜肝组织(包括肿瘤组织与癌旁组织)。采用流式细胞术检测肿瘤组织、癌旁组织、CD8⁺T细胞表达水平。根据术后是否复发,将68例肝癌患者分为复发组(25例)与非复发组(43例)。根据最终生存情况分为死亡组(18例)与生存组(50例)。统计并比较患者临床病理特征。采用Cox回归分析评估PLC切除术患者治疗后生存情况的影响因素。对上述所有研究对象进行随访,随访时间为2020年1月至2024年1月。采用Kaplan-Meier法,基于肝癌组织浸润淋巴细胞CD8⁺T细胞数量绘制生存曲线,Log-Rank检验比较生存曲线。结果复发组中,17例患者的CD8⁺T细胞数量≤100个/高倍视野,少于非复发组的12例(P<0.05)。复发组死亡12例,多于非复发组的6例(P<0.05)。死亡组与存活组中,肿瘤≥5 cm的患者均为14例,但差异有统计学差异(P<0.05)。死亡组13例肿瘤个数≥2个,少于存活组的21例(P<0.05)。死亡组11例有肿瘤包膜,少于存活组的13例(P<0.05)。死亡组与存活组中,有肿瘤血栓的患者均为9例,但差异有统计学意义(P<0.05)。死亡组术后1月,10例甲胎蛋白(AFP)＜400 ng/mL,少于存活组的31例(P<0.05)。死亡组5例Edmondson-Steiner分级为Ⅰ、Ⅱ级,少于存活组的33例(P<0.05)。死亡组6例AFP<20 ng/mL,少于存活组的31例(P<0.05)。肿瘤浸润淋巴细胞主要存在于癌旁组织间质中,肿瘤浸润细胞以CD8⁺T细胞为主,癌旁组织中CD8⁺T细胞数量低于肿瘤组织。Cox单因素及多因素风险比例回归分析显示,AFP水平(P=0.041)、CD8⁺T细胞数量(P=0.009)是PLC患者手术后复发情况的影响因素。Cox单因素风险比例回归分析显示,肿瘤大小(P=0.001)、肿瘤包膜(P=0.037)、术后1个月血清AFP(P=0.034)、Edmondson-Steiner分级(P=0.013)、AFP(P=0.013)是PLC患者手术后死亡的影响因素;多因素分析显示AFP(P=0.009)、CD8⁺T细胞数量(P=0.006)是PLC患者手术后死亡的影响因素。PLC患者手术后随访时间为4年,68例患者中,共存活50例,死亡18例;复发25例,非复发43例。Kaplan-Meier法与Log-Rank检验结果显示,浸润淋巴细胞CD8⁺T细胞数量与预后情况具有相关性,结果具有统计学意义(χ²=6.026,P=0.014),浸润淋巴细胞CD8⁺T细胞数量高者复发情况较少。结论 PLC患者肿瘤微环境中CD8⁺ T细胞浸润水平与患者预后密切相关,高水平的CD8⁺ T细胞浸润与较好的临床预后相关,提示CD8⁺ T细胞可能是肝癌治疗中一个重要的免疫靶标。

关键词: 原发性肝癌, CD8⁺细胞毒性T淋巴细胞, 肿瘤微环境, 预后, 流式细胞术

Abstract: Objective This study aims to explore the relationship between the degree of infiltration of CD8⁺ cytotoxic T lymphocytes in the tumor microenvironment of patients with primary liver cancer and their prognosis, with the hope of providing a new immunological basis for the treatment of liver cancer. Methods This study was retrospective, selecting 68 patients who underwent surgery for primary liver cancer between January 2018 and January 2020 as research subjects. Fresh liver tissues (including tumor and adjacent non-tumor tissues) were obtained during the surgery. Flow cytometry was used to assess the infiltration levels (IL) of CD8⁺ T cells in tumor tissues, adjacent non-tumor tissues. Patients were divided into recurrence (25 patients) and non-recurrence groups (43 patients) based on postoperative recurrence, and into survival (50 patients) and death groups (18 patients) based on final survival status. Clinical and pathological characteristics were statistically compared. All the above study subjects were followed up, with the follow-up period from January 2020 to January 2024. Cox regression analysis was utilized to examine factors affecting survival post-hepatectomy. Survival curves based on CD8⁺ T IL quantities were plotted using the Kaplan-Meier method, and differences were analyzed using the log-rank test. Results The recurrence group had 17 patients with ≤100 CD8⁺ T cells, fewer than the non-recurrence group′s 12 patients (P<0.05). The death group included 14 patients with tumors ≥5 cm, significantly more than the survival group (P<0.05), and 13 patients with ≥2 tumors, fewer than the 21 in the survival group (P<0.05). The death group also had 11 patients with encapsulated tumors, fewer than the survival group′s 13 patients (P<0.05), and 9 with tumor thrombi, the same number as the survival group but with statistical significance (P<0.05). Ten patients in the death group had postoperative alpha-fetoprotein (AFP) levels <400 ng/mL, fewer than the 31 in the survival group (P<0.05). Univariate and multivariate Cox regression analyses identified alpha-fetoprotein levels (P=0.041) and CD8⁺ T cell counts (P=0.009) as factors influencing recurrence post-surgery. Tumor size (P=0.001), capsule presence (P=0.037), serum AFP levels one month post-surgery (P=0.034), Edmondson-Steiner grading (P=0.013), and alpha-fetoprotein levels (P=0.013) were factors affecting mortality in univariate analysis, while AFP levels (P=0.009) and CD8⁺ T cell counts (P=0.006) were identified in multivariate analysis. Conclusion The level of CD8⁺ T cell infiltration in the tumor microenvironment of primary liver cancer patients is closely related to their prognosis. Higher levels of CD8⁺ T cell infiltration correlate with a better clinical outcome, suggesting that CD8⁺ T cells may be an important immunological target in liver cancer treatment.

Key words: Primary liver cancer, CD8⁺ cytotoxic T lymphocytes, Tumor microenvironment, Prognosis, Flow cytometry

范斌, 黄胜利, 易钢锋, 周宇. 原发性肝癌患者肿瘤微环境中CD8⁺细胞毒性T淋巴细胞浸润水平与预后的关系探究[J]. 肝脏, 2026, 31(3): 369-374.

FAN Bin, HUANG Sheng-li, YI Gang-feng, ZHOU Yu. Study on the relationship between prognosis in primary liver cancer patients and the infiltration levels of CD8⁺ cytotoxic T lymphocytes in the tumor microenvironment[J]. Chinese Hepatolgy, 2026, 31(3): 369-374.

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