结直肠癌组织NRF2蛋白表达与M1型TAMs的相关性及对患者预后的影响

Correlation between NRF2 protein expression and M1-type tumor-associated macrophages in colorectal cancer tissues and their effects on patient prognosis

摘要:

目的 探讨结直肠癌（CRC）组织中核转录因子红系2相关因子2（NRF2）蛋白表达与M1型肿瘤相关巨噬细胞（TAMs）的相关性，并分析其对CRC患者预后的影响。方法 回顾性分析2022年5月—2023年12月在新乡医学院洛阳市中心医院进行手术治疗的125例结直肠癌患者的临床资料。收集患者CRC组织。采用免疫组织化学方法检测CD86蛋白标记的CRC组织中的M1型TAMs的表达，根据CD86阳性细胞平均密度进行分组：高CD86阳性细胞组56例，低CD86阳性细胞组69例。采用Western blotting检测两组CRC组织中NRF2蛋白相对表达量。分析两组临床资料，采用Spearman法分析NRF2蛋白相对表达量与M1型TAMs的相关性。绘制生存曲线。采用多因素Cox比例风险回归模型分析CRC患者预后的影响因素。结果 高CD86组肿瘤大小<5 cm占比、分化程度中/低占比、CD8⁺肿瘤浸润性细胞毒性T淋巴细胞（CD8⁺ TILs）密度高占比均高于低CD86组（P <0.05）。高CD86组NRF2蛋白相对表达量高于低CD86组（P <0.05）。Spearman相关分析结果表明，肿瘤大小（r_s =-0.328，P =0.000）、分化程度（r_s =-0.237，P =0.008）与CD86水平呈负相关，CD8⁺ TILs密度（r_s =0.353，P =0.000）、NRF2蛋白水平（r_s =0.763，P =0.000）与CD86水平呈正相关。高CD86组预后总生存期长于低CD86组，高CD86组无病生存期长于低CD86组（均P <0.05）。多因素Cox比例风险回归分析结果表明：肿瘤大小≥5 cm［H^R =2.091（95% CI：1.012，4.319）］、分化程度中/低［H^R =2.685（95% CI：1.161，6.210）］均为影响CRC患者预后的危险因素（P <0.05）；CD8⁺ TILs密度高［H^R =0.433（95% CI：0.206，0.910）］、NRF2蛋白相对表达量高［H^R =0.713（95% CI：0.510，0.995）］均为影响CRC患者预后的保护因素（P <0.05）。结论 NRF2蛋白相对表达量与M1型TAMs水平呈正相关，是CRC患者预后的保护因素。

Abstract:

Objective To investigate the correlation between nuclear factor erythroid 2-related factor 2 (NRF2) protein expression in colorectal cancer tissues and M1-type tumor-associated macrophages (TAMs) and to analyze its effect on the prognosis of patients with colorectal cancer (CRC).Methods The clinical data of 125 patients with colorectal cancer who underwent surgery at Luoyang Central Hospital Affiliated to Xinxiang Medical University from May 2022 to December 2023 were retrospectively analyzed. CRC tissues were collected. Immunohistochemistry was used to detect M1-type TAMs in CRC tissues using CD86 as a marker. According to the mean density of CD86-positive cells, 56 patients were assigned to the high CD86-positive cell group and 69 to the low CD86-positive cell group. Western blotting was used to determine the relative expression of NRF2 protein in CRC tissues from the two groups. Clinical data were analyzed, Spearman correlation analysis was used to assess the correlation between relative NRF2 protein expression and M1-type TAMs, and survival curves were plotted.Results The proportions of tumors < 5 cm, moderate/poor differentiation, and high CD8⁺ tumor-infiltrating cytotoxic T lymphocyte (CD8⁺ TIL) density were higher in the high CD86 group than in the low CD86 group (P < 0.05). Relative NRF2 protein expression was higher in the high CD86 group than in the low CD86 group (P < 0.05). Spearman correlation analysis showed that tumor size (r_s = -0.328, P = 0.000) and degree of differentiation (r_s = -0.237, P = 0.008) were negatively correlated with CD86 levels, whereas CD8⁺ TIL density (r_s = 0.353, P = 0.000) and NRF2 protein levels (r_s = 0.763, P = 0.000) were positively correlated with CD86 levels. Overall survival and disease-free survival were longer in the high CD86 group than in the low CD86 group (both P < 0.05). Multivariate Cox proportional hazards regression analysis showed that tumor size ≥ 5 cm [H^R = 2.091 (95% CI: 1.012, 4.319) ] and moderate/poor differentiation [H^R = 2.685 (95% CI: 1.161, 6.210) ] were risk factors for the prognosis of patients with CRC (P < 0.05), whereas high CD8⁺ TIL density [H^R = 0.433 (95% CI: 0.206, 0.910) ] and high relative NRF2 protein expression [H^R = 0.713 (95% CI: 0.510, 0.995) ] were protective factors (P < 0.05).Conclusion Relative NRF2 protein expression is positively correlated with M1-type TAM levels and is a protective prognostic factor in patients with CRC.