腹腔镜结直肠癌根治患者手术前后外周血免疫细胞表达变化及其与短期预后的关系

Study on the relationship between changes of peripheral blood immune cell expression before and after operation and short-term prognosis in patients with laparoscopic radical resection for colorectal cancer

目的探讨结直肠癌患者腹腔镜根治性结直肠癌切除手术前后外周血中免疫细胞的变化,并进一步分析其与短期预后的关系。方法选择2021年3月至2022年3月该院收治的418例结直肠癌患者纳入结直肠癌组,136例门诊体检的健康志愿者纳入对照组,所有结直肠癌患者均接受腹腔镜根治性结直肠癌切除手术,术前和术后1、3、5 d采集患者外周静脉血,采用流式细胞仪检测外周血中CD3^(+)T淋巴细胞、CD4^(+)T淋巴细胞、CD8^(+)T淋巴细胞、CD4^(+)/CD8^(+)双阳性T淋巴细胞、CD19+B淋巴细胞,并计算CD4^(+)/CD8^(+)比值。术后随访至2023年3月31日,收集患者术后复发、转移和生存情况,将复发、转移定义为预后不良,无复发、转移定义为预后良好,分析预后不良组与预后良好组外周血免疫细胞的差异。结果结直肠癌组术前外周血CD3^(+)T淋巴细胞、CD4^(+)T淋巴细胞、CD4^(+)/CD8^(+)比值明显低于对照组,CD8^(+)T淋巴细胞、CD4^(+)CD8^(+)双阳性T淋巴细胞、CD19+B淋巴细胞明显高于对照组,差异有统计学意义(P<0.05)。结直肠癌患者术后外周血CD3^(+)T淋巴细胞、CD4^(+)T淋巴细胞、CD4^(+)/CD8^(+)比值先降低再升高,CD8^(+)T淋巴细胞、CD4^(+)CD8^(+)双阳性T淋巴细胞、CD19+B淋巴细胞先升高后降低,且术后5 d外周血CD3^(+)T淋巴细胞、CD4^(+)T淋巴细胞、CD4^(+)/CD8^(+)比值高于术后1 d,差异有统计学意义(P<0.05),术后5 d CD8^(+)T淋巴细胞、CD4^(+)CD8^(+)双阳性T淋巴细胞、CD19+B淋巴细胞低于术后1 d,差异有统计学意义(P<0.05)。所有患者随访19(12~24)个月,随访期间复发17例,转移65例,预后不良组术后5 d CD4^(+)/CD8^(+)比值低于预后良好组,CD4^(+)CD8^(+)双阳性T淋巴细胞、CD19+B淋巴细胞高于预后良好组,差异有统计学意义(P<0.05)。术后5 d低CD4^(+)/CD8^(+)比值、高CD4^(+)CD8^(+)双阳性T淋巴细胞、高CD19+B淋巴细胞结直肠癌患者无疾病进展生存率低于高CD4^(+)/CD8^(+)比值、低CD4^(+)CD8^(+)双阳性T淋巴细胞、低CD19+B淋巴细胞结直肠癌患者(P<0.05)。Cox回归分析结果显示TNM分期Ⅲ期、淋巴结转移、术后放疗、术后5 d CD4^(+)CD8^(+)双阳性T淋巴细胞升高、术后5 d CD19+B淋巴细胞升高是结直肠癌患者预后不良的危险因素(P<0.05),术后5 d CD4^(+)/CD8^(+)比值升高是结直肠癌患者预后不良保护因素(P<0.05)。结论结直肠癌患者术后低CD4^(+)/CD8^(+)比值、高CD4^(+)CD8^(+)双阳性T淋巴细胞、高CD19+B淋巴细胞与术后短期内不良预后有关,可作为结直肠癌术后预后评估的指标。

Objective To investigate the expression of immune cells in peripheral blood of patients with colorectal cancer before and after laparoscopic radical resection of colorectal cancer and its relationship with short-term prognosis.Methods A total of 418 patients with colorectal cancer admitted to the First Affiliated Hospital of Chongqing Medical University from March 2021 to March 2022 were included in the colorectal cancer group,and 136 healthy volunteers in the outpatient department were included in the control group.All patients with colorectal cancer received laparoscopic radical resection of colorectal cancer.Venous blood was collected on preoperative,postoperative 1,3,5 d,and flow cytometry was used to detect CD3^(+)T lymphocytes,CD4^(+)T lymphocytes,CD8^(+)T lymphocytes,CD4^(+)CD8^(+)double positive T lymphocytes and CD19+B lymphocytes in peripheral blood,and CD4^(+)/CD8^(+)ratio was calculated.The patients were followed up until March 31,2023,and their status of postoperative recurrence,metastasis and survival were collected.The recurrence and metastasis were defined as poor prognosis,no recurrence and metastasis were defined as good prognosis.The differences in peripheral blood immune cells between the poor prognosis group and the good prognosis group were analyzed.Results The ratios of CD3^(+)T lymphocytes,CD4^(+)T lymphocytes and CD4^(+)/CD8^(+)preoperative in peripheral blood of colorectal cancer group were lower than those of control group,CD8^(+)T lymphocytes,CD4^(+)CD8^(+)double positive T lymphocytes and CD19+B lymphocytes were higher than those of control group,the differences were significant(P<0.05).The ratio of CD3^(+)T lymphocytes,CD4^(+)T lymphocytes and CD4^(+)/CD8^(+)in peripheral blood of patients with colorectal cancer after surgery decreased first and then increased,while CD8^(+)T lymphocytes,CD4^(+)CD8^(+)double positive T lymphocytes and CD19+B lymphocytes increased first then decreased.The CD3^(+)T lymphocytes,CD4^(+)T lymphocytes and CD4^(+)/CD8^(+)ratio in peripheral blood 5 days after operation were higher than those 1 day after operation,the differences were statistically significant(P<0.05).CD8^(+)T lymphocytes,CD4^(+)CD8^(+)double positive T lymphocytes and CD19+B lymphocytes on the 5 days after operation were lower than those on 1 d after operation(P<0.05).The duration follow-up was 19(12—24)months,during which there were 17 cases of relapse,65 cases of metastasis.The CD4^(+)/CD8^(+)ratio at 5 days after operation in the poor prognosis group was lower than that in the good prognosis group(P<0.05),CD4^(+)CD8^(+)double positive T lymphocytes and CD19+B lymphocytes were higher than those in the good prognosis group(P<0.05).The rates of progression-free survival in colorectal cancer patients with low CD4^(+)/CD8^(+)ratio,high CD4^(+)CD8^(+)double-positive T lymphocytes and high CD19+B lymphocytes at 5 days after operation were lower than those in colorectal cancer patients with high CD4^(+)/CD8^(+)ratio,low CD4^(+)CD8^(+)double-positive T lymphocytes and low CD19+B lymphocytes(P<0.05).Cox regression analysis showed that TNM stageⅢ,lymph node metastasis,postoperative radiotherapy,high CD4^(+)CD8^(+)double positive T lymphocytes at 5 days after operation and high CD19+B lymphocytes 5 days after operation were risk factors for poor prognosis in colorectal cancer patients(P<0.05),and high CD4^(+)/CD8^(+)ratio at 5 days after operation was protective factor(P<0.05).Conclusion Low CD4^(+)/CD8^(+)ratio,high CD4^(+)CD8^(+)double positive T lymphocytes and high CD19+B lymphocytes associate with poor prognosis in the short term after surgery in patients with colorectal cancer.