结直肠癌组织高表达IL-35对Th1和Th17可塑性的作用

doi:10.3969/j.issn.1673-8640.2024.04.006

摘要/Abstract

摘要：

目的探讨结直肠癌组织中高表达的抑制性细胞因子白细胞介素（IL）-35对辅助性T细胞1（Th1）和辅助性T细胞17（Th17）可塑性的作用。方法选取2019年3月—2020年2月上海交通大学医学院附属新华医院结直肠癌患者90例（结直肠癌组）和健康体检者28名（正常对照组）。采用流式细胞术检测结直肠癌组和正常对照组外周血Th1百分比（Th1%）和Th17百分比（Th17%）。基于Kaplan-Meier Plotter数据库分析γ-干扰素（IFN-γ）和IL-17A高表达对结直肠癌患者总生存期和预后不良的影响。采用免疫组化法检测结直肠癌患者癌组织和癌旁组织（距肿瘤边缘≥2 cm）中IL-35的表达。采用酶联免疫吸附试验检测结直肠癌患者和正常对照者血清IL-35水平。采用体外诱导Th1分化实验观察IL-35对Th1分泌IL-17A和IFN-γ的影响。结果与正常对照组比较，结直肠癌组Th1%显著降低（P=0.019），Th17%显著升高（P<0.001）。结直肠癌组Th1%与Th17%的r²值（0.393 4）与正常对照组（0.041 0）比较差异有统计学意义（P=0.007）。基于Kaplan-Meier Plotter数据库的分析结果显示，IFN-γ mRNA低表达是结直肠癌患者总生存期缩短的危险因素[风险比（HR）=0.74，95%可信区间（CI）为0.58~0.94，P=0.013]，IL-17A mRNA高表达是结直肠癌患者总生存期缩短的危险因素（HR=1.26，95%CI为1.02~1.59，P=0.020）。结直肠癌患者癌组织IL-35表达显著高于癌旁组织（P<0.001）。Ⅰ~Ⅱ期（早期）和Ⅲ~Ⅳ期（晚期）结直肠癌患者血清IL-35水平均显著高于正常对照者（P<0.05）。体外诱导Th1分化实验结果显示，IL-35可降低Th1中IFN-γ表达，提高IL-17A表达。结论结直肠癌患者外周血Th1和Th17呈异常变化，且IL-35表达增加。高表达的IL-35可降低Th1中IFN-γ表达，提高IL-17A表达。

关键词: 白细胞介素-35, 结直肠癌, 辅助性T细胞1, 辅助性T细胞17, 肿瘤免疫

Abstract:

Objective To investigate the effect of highly expressed inhibitory cytokine interleukin（IL）-35 on the plasticity of T helper cell 1（Th1） and T helper cell 17（Th17）in colorectal cancer tissues. Methods A total of 90 patients with colorectal cancer（colorectal cancer group） and 28 healthy subjects（healthy control group） were enrolled from Xinhua Hospital of Shanghai Jiao Tong University School of Medicine from March 2019 to February 2020. Flow cytometry was used to determine Th1 percentage（Th1%） and Th17 percentage（Th17%） in peripheral blood of colorectal cancer group and healthy control group. The effects of highly expressed interferon-gamma（IFN-γ） and IL-17A on overall survival and poor prognosis of colorectal cancer were analyzed based on Kaplan-Meier Plotter database. Immunohistochemistry was used to determine the expression of IL-35 in cancer tissues and adjacent tissues（2 cm from tumor margin） of colorectal cancer patients. Serum IL-35 levels in colorectal cancer and healthy control groups were determined by enzyme-linked immunosorbent assay. The effect of IL-35 on the secretion of IL-17A and IFN-γ by Th1 was observed based on the induced Th1 differentiation induction experiment in vitro. Results Compared with healthy control group，Th1% in colorectal cancer group was decreased（P=0.019），and Th17% was increased（P<0.001）. The r² values of Th1% and Th17% in colorectal cancer group（0.393 4） were different from those in healthy control group（0.041 0）（P=0.007）. Low expression of IFN-γ mRNA was a risk factor for shorter survival in colorectal cancer patients [hazard ratio（HR）=0.74，95% confidence interval（CI）0.58-0.94，P=0.013]. High expression of IL-17A mRNA was a risk factor for shorter survival in colorectal cancer patients（HR=1.26，95%CI 1.02-1.59，P=0.020）. The expression of IL-35 in colorectal cancer tissues was higher than that in adjacent tissues（P<0.001）. Serum IL-35 levels in patients with stage Ⅰ to Ⅱ（early stage） and stage Ⅲ to Ⅳ（advanced stage） colorectal cancer were higher than those in healthy control group（P<0.05）. According to the results of Th1 differentiation induction experiment in vitro，IL-35 can decrease the expression of IFN-γ and increase the expression of IL-17A in Th1. Conclusions The proportions of Th1 and Th17 in peripheral blood of patients with colorectal cancer show abnormal changes，and the expression of IL-35 increases. High expression of IL-35 can decrease the expression of Th1 IFN-γ and increase the expression of IL-17A.

Key words: Interleukin-35, Colorectal cancer, T helper cell 1, T helper cell 17, Tumor immunity

中图分类号:

R446.1

许耘川, 皇甫昱婵, 马妍慧. 结直肠癌组织高表达IL-35对Th1和Th17可塑性的作用[J]. 检验医学, 2024, 39(4): 343-350.

XU Yunchuan, HUANGFU Yuchan, MA Yanhui. Effect of highly expressed IL-35 on Th1 and Th17 plasticity in colorectal cancer[J]. Laboratory Medicine, 2024, 39(4): 343-350.

图/表 10

参考文献 23

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TNM分期			年龄				性别				病理类型
TNM分期			≤60岁		>60岁		男		女		管状腺癌	黏液性腺癌	乳头状腺癌	未知
Ⅰ~Ⅱ期	（54例）		7（12.96）		47（87.03）		28（51.85）		26（48.15）		44（81.58）	3（5.26）	3（5.26）	4（7.89）
Ⅲ~Ⅳ期	（36例）		9（25.00）		27（75.00）		19（52.78）		17（47.22）		27（75.00）	3（8.33）	2（5.56）	4（11.11）
U值			871.00				963.00				772.50
P值			0.41				0.93				0.70
TNM分期		肿瘤部位
TNM分期		升结肠		横结肠		降结肠		乙状结肠		直肠
Ⅰ~Ⅱ期		7（12.96）		16（29.63）		4（7.41）		22（40.74）		5（9.26）
Ⅲ~Ⅳ期		6（16.67）		14（38.89）		5（13.89）		10（27.78）		1（2.78）
U值		787.50
P值		0.11

TNM分期			年龄				性别				病理类型
TNM分期			≤60岁		>60岁		男		女		管状腺癌	黏液性腺癌	乳头状腺癌	未知
Ⅰ~Ⅱ期	（54例）		7（12.96）		47（87.03）		28（51.85）		26（48.15）		44（81.58）	3（5.26）	3（5.26）	4（7.89）
Ⅲ~Ⅳ期	（36例）		9（25.00）		27（75.00）		19（52.78）		17（47.22）		27（75.00）	3（8.33）	2（5.56）	4（11.11）
U值			871.00				963.00				772.50
P值			0.41				0.93				0.70
TNM分期		肿瘤部位
TNM分期		升结肠		横结肠		降结肠		乙状结肠		直肠
Ⅰ~Ⅱ期		7（12.96）		16（29.63）		4（7.41）		22（40.74）		5（9.26）
Ⅲ~Ⅳ期		6（16.67）		14（38.89）		5（13.89）		10（27.78）		1（2.78）
U值		787.50
P值		0.11

组别	例数	Th1%/%	Th17%/%
结直肠癌组	31	10.38（4.03，15.38）	2.16（1.56，3.12）
正常对照组	28	16.75（12.37，21.89）	0.96（0.76，1.83）
U值		280	167
P值		0.019	<0.001

组别	例数	Th1%/%	Th17%/%
结直肠癌组	31	10.38（4.03，15.38）	2.16（1.56，3.12）
正常对照组	28	16.75（12.37，21.89）	0.96（0.76，1.83）
U值		280	167
P值		0.019	<0.001

临床病理特征	例数	癌组织			癌旁组织			临床病理特征	例数	癌组织			癌旁组织
临床病理特征	例数	低表达	高表达	缺失	低表达	高表达	缺失	临床病理特征	例数	低表达	高表达	缺失	低表达	高表达	缺失
年龄								T分期
≤60岁	34	15	18	1	30	2	2	T1/T2期	9	6	3	0	8	1	0
>60岁	54	36	18	0	49	3	2	T3/T4期	79	46	32	1	72	4	3
缺失	2	1	1	0	2	0	0	缺失	2	0	2	0	1	0	1
P值			0.053			0.928		P值			0.685			0.483
性别								淋巴转移
男	47	27	20	0	43	2	2	N0	56	33	23	0	51	2	3
女	43	25	17	0	38	3	2	N1/N2	34	19	14	1	30	3	1
P值			0.844			0.572		P值			0.901			0.308
肿瘤大小								远隔转移
≤5 cm	46	25	20	1	40	4	2	M0	87	51	35	1	79	4	4
>5 cm	42	16	26	9	39	1	2	M1	2	1	1	0	1	1	0
缺失	2	1	1	0	2	0	0	缺失	1	0	1	0	1	0	0
P值			0.550			0.205		P值			0.793			0.008
病理分级								TNM分期
Ⅰ级	5	5	0	0	5	0	0	Ⅰ~Ⅱ期	54	33	21	0	50	2	2
Ⅱ~Ⅲ级	85	47	37	1	76	5	4	Ⅲ~Ⅳ期	34	19	14	1	30	3	1
P值			0.054			0.569		缺失	2	0	2	0	1	0	1
								P值			0.746			0.319