ACREM Asian Case Reports in Emergency Medicine 2328-0409 Scientific Research Publishing 10.12677/ACREM.2024.121001 ACREM-80406 ACREM20240100000_47229416.pdf 医药卫生 非小细胞肺癌患者Th17细胞、血清IL-17、MMP2的表达在临床病理分期及淋巴结转移的诊断价值 The Diagnostic Value of Th17 Cells, Serum IL-17, and MMP2 Expression in Clinical Pathological Staging and Lymph Node Metastasis in Non-Small Cell Lung Cancer Patients 刘 延梅 1 2 宋 荟琴 3 1 黄 晓燕 4 1 陕西省人民医院陕西省感染与免疫疾病重点实验室，陕西 西安 陕西省铜川矿务局中心医院检验科，陕西 铜川 null 陕西省人民医院西院二病区，陕西 西安 02 02 2024 12 01 1 7 © Copyright 2014 by authors and Scientific Research Publishing Inc. 2014 This work is licensed under the Creative Commons Attribution International License (CC BY). http://creativecommons.org/licenses/by/4.0/

目的：探讨Th17细胞、血清白细胞介素-17 (IL-17)、基质金属蛋白酶(MMP2)在非小细胞肺癌(NSCLC)临床分期及淋巴结转移中的临床应用价值。方法：选取2022年11月~2023年10月入住于陕西省人民医院的42例NSCLC患者(NSCLC组)及40例肺良性病变患者(对照组)，采用流式细胞术检测各组外周血Th17细胞水平，采用酶联免疫吸附法(ELISA)测定血清中IL-17和MMP2水平，评估这三个检测指标在NSCLC临床分期和淋巴结转移中的应用价值及三者联合检测的诊断效能。结果：NSCLC组中，与I~II期患者对比，III~IV期患者外周血Th17细胞水平明显增高，血清IL-17和MMP2水平也明显较高，差异均有统计学意义(均p < 0.05)；NSCLC组中，淋巴结转移者Th细胞水平、血清IL-17和MMP-2水平与未转移者对比，明显较高，差异均有统计学意义(均p < 0.05)；Th17，IL-17和MMP2三项联合检测准确度、特异度、灵敏度分别为95.71%，93.84%和97.22%，与单项检测对比，均明显较高，差异均有统计学意义(均p < 0.05)。结论：Th17细胞，IL-17和MMP2在NSCLC与NSCLC临床分期及淋巴结转移中有一定关联，三者联合检测，有助于NSCLC的鉴别诊断。 Objective: To explore the clinical application value of Th17 cells, serum interleukin-17 (IL-17), and matrix metalloproteinases (MMP2) in the clinical staging and lymph node metastasis of non-small cell lung cancer (NSCLC). Methods: 42 NSCLC patients (NSCLC group) and 40 patients with benign lung lesions (control group) admitted to Shaanxi Provincial People’s Hospital from November 2022 to October 2023 were se-lected. Flow cytometry was used to detect peripheral blood Th17 cell levels in each group, and en-zyme-linked immunosorbent assay (ELISA) was used to measure IL-17 and MMP2 levels in the se-rum. The application value of these three detection indicators in clinical staging and lymph node metastasis of NSCLC, as well as the diagnostic efficacy of their combined detection were evaluated. Results: In the NSCLC group, compared with patients in stages I~II, patients in stages III~IV showed a significant increase in peripheral blood Th17 cell levels, as well as significantly higher serum IL-17 and MMP2 levels, with statistical significance (all p < 0.05); in the NSCLC group, the levels of Th cells, serum IL-17, and MMP-2 were significantly higher in patients with lymph node metastasis compared to those without metastasis, and the differences were statistically significant (all p < 0.05); the accuracy, specificity, and sensitivity of the combined detection of Th17, IL-17, and MMP2 were 95.71%, 93.84%, and 97.22%, respectively. Compared with single detection, they were significantly higher, and the differences were statistically significant (all p < 0.05). Conclusion: Th17 cells, IL-17, and MMP2 are associated with clinical staging and lymph node metastasis in NSCLC, and their combined detection can help in the differential diagnosis of NSCLC.

 非小细胞肺癌，Th17细胞，IL-17，MMP2, Non-Small Cell Lung Cancer Th17 Cells Serum Interleukin-17 Matrix Metalloproteinases 摘要

目的：探讨Th17细胞、血清白细胞介素-17 (IL-17)、基质金属蛋白酶(MMP2)在非小细胞肺癌(NSCLC)临床分期及淋巴结转移中的临床应用价值。方法：选取2022年11月~2023年10月入住于陕西省人民医院的42例NSCLC患者(NSCLC组)及40例肺良性病变患者(对照组)，采用流式细胞术检测各组外周血Th17细胞水平，采用酶联免疫吸附法(ELISA)测定血清中IL-17和MMP2水平，评估这三个检测指标在NSCLC临床分期和淋巴结转移中的应用价值及三者联合检测的诊断效能。结果：NSCLC组中，与I~II期患者对比，III~IV期患者外周血Th17细胞水平明显增高，血清IL-17和MMP2水平也明显较高，差异均有统计学意义(均p < 0.05)；NSCLC组中，淋巴结转移者Th细胞水平、血清IL-17和MMP-2水平与未转移者对比，明显较高，差异均有统计学意义(均p < 0.05)；Th17，IL-17和MMP2三项联合检测准确度、特异度、灵敏度分别为95.71%，93.84%和97.22%，与单项检测对比，均明显较高，差异均有统计学意义(均p < 0.05)。结论：Th17细胞，IL-17和MMP2在NSCLC与NSCLC临床分期及淋巴结转移中有一定关联，三者联合检测，有助于NSCLC的鉴别诊断。

 关键词

非小细胞肺癌，Th17细胞，IL-17，MMP2

 The Diagnostic Value of Th17 Cells, Serum IL-17, and MMP2 Expression in Clinical Pathological Staging and Lymph Node Metastasis in Non-Small Cell Lung Cancer Patients<sup> </sup>

Yanmei Liu1, Huiqin Song2, Xiaoyan Huang3*

1West Hospital Second Ward, Shaanxi Provincial People’s Hospital, Xi’an Shaanxi

2Laboratory Department, Central Hospital of Tongchuan Mining Bureau in Shaanxi Province, Tongchuan Shaanxi

3Key Laboratory of Infectious and Immunological Diseases of Shaanxi Provincial People’s Hospital, Xi’an Shaanxi

Received: Dec. 13th, 2023; accepted: Jan. 26th, 2024; published: Feb. 2nd, 2024

 ABSTRACT

Objective: To explore the clinical application value of Th17 cells, serum interleukin-17 (IL-17), and matrix metalloproteinases (MMP2) in the clinical staging and lymph node metastasis of non-small cell lung cancer (NSCLC). Methods: 42 NSCLC patients (NSCLC group) and 40 patients with benign lung lesions (control group) admitted to Shaanxi Provincial People’s Hospital from November 2022 to October 2023 were selected. Flow cytometry was used to detect peripheral blood Th17 cell levels in each group, and enzyme-linked immunosorbent assay (ELISA) was used to measure IL-17 and MMP2 levels in the serum. The application value of these three detection indicators in clinical staging and lymph node metastasis of NSCLC, as well as the diagnostic efficacy of their combined detection were evaluated. Results: In the NSCLC group, compared with patients in stages I~II, patients in stages III~IV showed a significant increase in peripheral blood Th17 cell levels, as well as significantly higher serum IL-17 and MMP2 levels, with statistical significance (all p < 0.05); in the NSCLC group, the levels of Th cells, serum IL-17, and MMP-2 were significantly higher in patients with lymph node metastasis compared to those without metastasis, and the differences were statistically significant (all p < 0.05); the accuracy, specificity, and sensitivity of the combined detection of Th17, IL-17, and MMP2 were 95.71%, 93.84%, and 97.22%, respectively. Compared with single detection, they were significantly higher, and the differences were statistically significant (all p < 0.05). Conclusion: Th17 cells, IL-17, and MMP2 are associated with clinical staging and lymph node metastasis in NSCLC, and their combined detection can help in the differential diagnosis of NSCLC.

Keywords:Non-Small Cell Lung Cancer, Th17 Cells, Serum Interleukin-17, Matrix Metalloproteinases

Copyright © 2024 by author(s) and beplay安卓登录

This work is licensed under the Creative Commons Attribution International License (CC BY 4.0).

http://creativecommons.org/licenses/by/4.0/

 1. 引言

肺癌是公认的发病率高、预后差的恶性肿瘤，对人类的生命健康构成了巨大的威胁 [ 1 ] 。每年死于肺癌的人数预计在160万人左右，其中有85%的肺癌在组织学上的分类为非小细胞肺癌(nonsmall cell lung cancer, NSCLC)，由于NSCLC早期无典型症状及体征，发现及确诊时已到中晚期 [ 2 ] 。目前NSCLC治疗的主要策略包括手术、放疗和化疗。手术和放疗通常用于切除肿瘤病灶，而化疗在临床实践中主要作为术后辅助治疗 [ 3 ] ，但适宜手术治疗者仅为20%~30%，术后仍有60%患者存活不满5年，追其原因是25%~50%的病例会在疾病转归过程中发生转移 [ 4 ] 。因此，在临床分期和转移前提高诊断率，简化诊断方式，推广无创有效的诊断方法对NSCLC患者的早期诊断、预防治疗等有重要意义。

近年的研究中发现一种新型的CD4+效应T细胞即Th17细胞参与了恶性肿瘤的病理发展过程 [ 5 ] 。白细胞介素-17 (interleukin-17, IL-17)是一种前炎性细胞因子，由Th17细胞分泌，具有很强的募集中性粒细胞、促进炎性因子释放作用，通过与IL-17受体结合，使下游基因表达细胞因子 [ 6 ] ，且可通过Th17参与细胞免疫，IL-17与肿瘤的发生发展密切相关 [ 7 ] 。有科学家在鼠模型体内发现，IL-17等Th17细胞相关因子可能参与了NSCLC的发生发展过程 [ 8 ] 。基质金属蛋白酶(Matrix metalloproteinase, MMP)中的MMP-2是机体中主要的IV型胶原降解酶，在肿瘤的形成过程中起重要作用 [ 9 ] 。本研究拟探讨Th17细胞、IL-17、MMP2在NSCLC临床分期及淋巴结转移中的临床应用价值，及评估三者联合检测的效能。

 2. 材料与方法 2.1. 研究对象

选取陕西省人民医院2022年11月~2023年10月经病理检验确诊的42例NSCLC患者(NSCLC组)及40例肺良性病变患者(良性病变组)为观察对象。NSCLC组纳入标准 [ 10 ] ：① 患者符合NSCLC诊断标准，且经病理检验确诊；② 患者心、肝、肾、胃功能正常；③ 入组前未接受相关放疗、化疗、免疫治疗；④ 预计生存期超3个月；⑤ 患者临床资料完整。NSCLC组排除标准：① 临床资料不全者；② 患有慢性阻塞性肺病、支气管哮喘等急慢性感染。③ 伴多发性硬化症、红斑狼疮、类风湿性关节炎等自身免疫性疾病；④ 并发其他恶性肿瘤；⑤ 妊娠期、哺乳期女性。NSCLC组：男性30例，女性12例，年龄22~65岁，平均年龄48.03 ± 5.14岁。腺癌28例，鳞癌14例。TNM分期：I期8例，II期12例，III期15例，IV期7例。淋巴结转移：有27例，无15例。良性病变组患者入组前均经CT与病理确诊，排除癌前病变，男性22例，女性18例，年龄27~65岁，平均年龄46.14 ± 4.28岁。两组患者性别、年龄对比差异无统计学意义(p > 0.05)，有可比性。以上患者均知情同意并经陕西省人民医院伦理委员会批准。

 2.2. 仪器与试剂

采用流式细胞术检测各组外周血Th17细胞水平，采用酶联免疫吸附法(ELISA)测定血清中IL-17和MMP2水平，ELISA试剂盒由武汉伊莱瑞特有限公司提供，所有操作严格按照试剂盒说明书进行。

 2.3. 方法

流式细胞检测：空腹抽取患者每人肘静脉血6 ml，其中肝素抗凝血3 mL，用于流式细胞检测；促凝剂处理的血液3 mL，用于ELISA检测。肝素抗凝血采用淋巴细胞分离液密度梯度离心(2000 r/min离心20 min)分离单个核细胞。每组取5 × 104细胞移入至流式管，向流式管中加入荧光抗体FITC-CD3和PerCP-CD4各10 μL，室温避光反应15 min。1500 r/min速率离心5 min，弃上清。在沉淀中加入Fixation/pormeabilization破膜剂1 mL，混匀室温放置20 min。加入PBS洗液2 mL，1500 r/min速率离心5 min，弃去上清液，留下沉淀。加入10 μL荧光标记的PE-IL17进行细胞内染色，避光反应30 min。加入PBS洗液2 mL，1500 r/min速率离心5 min，弃去上清液，留下沉淀。如此重复洗涤两次后，加入500 μL PBS重悬细胞进行流式检测。

ELISA检测：促凝剂处理的血液常温静置2 h，按2500 r/min离心10 min，分离血清。配置不同浓度标准品，然后将标本和不同浓度标准品加入相应板孔，再加入相应的生物标记素进行标记，置于37℃孵育60 min。洗板4次后，分别加入抗人IL-17和MMP2生物素化的抗体进行反应，37℃孵育60 min，加入特异性酶溶液作用后洗板，加入显色液终止反应，测其OD值，通过绘制标准曲线计算得出IL-17和MMP2的含量。

 2.4. 观察指标

观察指标如下：① 比较Th17，IL-17和MMP2在NSCLC组和良性病变组中的表达。② 比较NSCLC组内I~II期和III~IV期患者Th17、IL-17和MMP表达情况。③ 比较NSCLC组内淋巴结转移和无转移患者Th17、IL-17和MMP2表达情况。④ 计算Th17、IL-17和MMP2单项诊断及三项联合诊断准确度、特异度、敏感度，其中准确度 = (真阳性 + 真阴性)/总例数 × 100%；灵敏度 = 真阳性/(真阳性 + 假阴性 × 100%。特异度 = 真阴性/(真阴性 + 假阳性) × 100%。

 2.5. 统计学分析

将研究数据纳入SPSS22.0统计学软件中，计数资料用百分率(%)表示，使用χ2检验；计量资料用均数 ± 标准差(x ± s)表示，使用方差分析进行组间比较，经t检验，p < 0.05为差异有统计学意义。

 3. 结果

1) NSCLC组和良性病变组血清CA724，NSE和PGR水平比较见表1。NSCLC组患者CA724，NSE和PGR水平与肺良性病变组对比，两组间差异均具有统计学意义(p < 0.05)。

Comparison of Th17, IL-17 and MMP2 levels between NSCLC group and benign lesion group (x ± s
检测指标 NSCLC组 良性病变组 t p
n = 42 n = 40
Th17细胞(%) 3.21 ± 0.88 1.00 ± 0.28 7.64 <0.001
IL-17 (pg/mL) 8.06 ± 1.57 2.68 ± 0.61 10.22 <0.001
MMP2 (pg/mL) 50.47 ± 3.46 18.79 ± 1.57 12.74 <0.001

表1. NSCLC组和良性病变组患者Th17、IL-17和MMP2水平比较(x ± s)

2) 不同NSCLC临床病理分期患者Th17、IL-17和MMP2水平比较见表2。NSCLC组中，III~IV期患者血清CA724，NSE和PGR水平与I~II 期患者比较，两组间差异具有统计学意义(p < 0.05)。

Comparison of Th17, IL-17 and MMP2 levels in patients with different clinicopathological stages of NSCLC (x ± s
检测指标 I~II期 III~IV期 t p
n = 20 n = 22
Th17细胞(%) 2.14 ± 0.93 5.07 ± 1.65 8.45 <0.001
IL-17 (pg/mL) 5.41 ± 1.08 10.23 ± 1.85 13.04 <0.001
MMP2 (pg/mL) 42.61 ± 2.81 68.04 ± 5.23 7.32 <0.001

表2. 不同NSCLC临床病理分期患者Th17、IL-17和MMP2水平比较(x ± s)

3) NSCLC淋巴结转移组患者与无淋巴结转移组患者血清Th17、IL-17和MMP2水平比较见表3。NSCLC组中淋巴结转移组患者Th17、IL-17和MMP2水平与无淋巴结转移组患者比较，两组间差异具有统计学意义(p < 0.05)。

Comparison of Th17, IL-17 and MMP2 levels between NSCLC lymph node metastasis group and non-lymph node metastasis group (x ± s
检测指标 无淋巴结转移 淋巴结转移 t p
n = 15 n = 27
Th17细胞(%) 2.19 ± 0.68 4.94 ± 1.35 6.33 <0.001
IL-17 (pg/mL) 4.94 ± 0.83 12.17 ± 1.99 17.12 <0.001
MMP2 (pg/mL) 39.41 ± 2.81 76.44 ± 6.02 14.30 <0.001

表3. NSCLC淋巴结转移组与无淋巴结转移组Th17、IL-17和MMP2水平比较(x ± s)

4) Th17、IL-17和MMP2单项及三项联合诊断效能比较见表4。Th17、IL-17和MMP2联合诊断准确度、特异度、灵敏度与单项检测比较，两组间差异有统计学意义(p < 0.05)。

Comparison of accuracy, specificity and sensitivity of Th17, IL-17 and MMP2 single and combined detection (%
指标 Th17 IL-17 MMP2 三项联合 三项联合与Th17 三项联合与IL-17 三项联合与MMP2
χ2 p χ2 p χ2 p
准确度 72.66 78.32 75.57 95.71 15.71 <0.001 18.57 <0.001 21.40 <0.001
特异度 75.48 76.74 72.11 93.84 5.78 0.024 8.43 0.006 6.12 0.004
灵敏度 80.22 82.05 75.48 97.22 10.24 0.003 16.07 <0.001 12.55 0.002

表4. Th17、IL-17和MMP2单项及三项联合检测准确度、特异度、灵敏度比较(%)

 4. 讨论

NSCLC是一种临床常见的肺部恶性肿瘤，占所有肺癌的80%，具有高发病率和病死率特点 [ 11 ] ，且近年来呈上升趋势，严重影响患者生命健康 [ 12 ] 。目前手术切除、放疗、化疗和联合治疗作为NSCLC的主要治疗手段，显著提高了NSCLC患者的生存率 [ 13 ] 。然而，NSCLC发病隐匿，早期临床体征不明显，由于缺乏有效的早期诊断标记物，往往确诊时已进入中晚期，且25%~40%的病例会在疾病转归过程中发生转移 [ 14 ] 。目前，临床症状和影像学检查对于微小病灶等的检出效果较一般，仅60%~70%，病理学检查为诊断金标准，但有一定创伤性 [ 15 ] ，因此，开发NSCLC早期的诊断性生物标志物和治疗靶点尤为重要。

本研究中，我们通过流式细胞术检测NSCLC和肺良性病变患者外周血Th17细胞水平，结果显示NSCLC患者外周血中Th17细胞水平明显高于良性病变组，且在NSCLC组中，III~IV期及淋巴结转移患者Th17细胞水平明显升高。这表明Th17细胞容易趋化到肺癌组织内，且在肺癌组织局部呈高表达。有研究发现肿瘤细胞与肿瘤成纤维细胞能分泌趋化因子RANTES与单核细胞趋化蛋白1 (MCP-1)，诱导Th17细胞在肿瘤局部聚集增殖 [ 16 ] ；此外，肿瘤微环境中肿瘤细胞会分泌一些促进Th17细胞增殖所需要的细胞因子，这对Th17细胞的分化有着至关重要的促进作用，其中以分泌的IL-17最常见 [ 17 ] 。这验证了本研究中IL-17结果，即NSCLC患者外周血中IL-17水平明显高于良性病变组，且在NSCLC组中，III~IV期及淋巴结转移患者IL-17水平明显升高。IL-17与炎性反应和免疫活动有密切关系，有学者认为IL-17可能通过趋炎反应促进局部肿瘤的血管生成及侵袭转移，也可诱导PI3K/AKT通路 [ 18 ] ，使抗凋亡 [ 19 ] 、促血管生成等基因的表达上调 [ 20 ] ，促肿瘤发生发展。

MMP2与肿瘤的浸润和转移密切相关，由肿瘤细胞和间质细胞分泌，参与降解3型胶原、纤维黏连蛋白等胞外基质成分 [ 21 ] 。肿瘤细胞的持续性生长和转移均有MMP2的参与，MMP2是肿瘤新生血管形成的关键 [ 22 ] ，其可促进VEGF的表达 [ 23 ] ，从而使血管内皮细胞增殖，使血管通透性增加 [ 24 ] ，使血管内皮细胞外基质降解，进而为血管生长及延伸提供基础 [ 25 ] 。这完全解释了我们研究结果中NSCLC患者外周血中MMP2水平明显高于良性病变组，且在III~IV期及淋巴结转移患者中明显升高。

同时，本研究结果提示，Th17、IL-17和MMP2联合检测，在NSCLC诊断中的准确度、特异度及灵敏度分别95.71%、93.84%和97.22%，均明显高于单一检测，提示三种指标联合检测的诊断效能较好。

 5. 结论

综上所述，Th17、IL-17和MMP2在NSCLC患者中表达异常，并能反映NSCLC临床分期及淋巴结转移程度，且三项联合检测具有较高诊断效能，可为NSCLC诊治提供可靠参考。

 基金项目

陕西省人民医院科技发展孵化基金项目(2021YJY-29)，陕西省人民医院菁英人才项目(2021JY-48)。

 文章引用

刘延梅,宋荟琴,黄晓燕. 非小细胞肺癌患者Th17细胞、血清IL-17、MMP2的表达在临床病理分期及淋巴结转移的诊断价值 The Diagnostic Value of Th17 Cells, Serum IL-17, and MMP2 Expression in Clinical Pathological Staging and Lymph Node Metastasis in Non-Small Cell Lung Cancer Patients[J]. 亚洲急诊医学病例研究, 2024, 12(01): 1-7. https://doi.org/10.12677/ACREM.2024.121001

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