具核梭杆菌感染对结肠癌转化生长因子-β1表达及肿瘤浸润淋巴细胞的影响

doi:10.3969/j.issn.1006-5725.2019.16.011

实用医学杂志 ›› 2019, Vol. 35 ›› Issue (16): 2564-2569.doi: 10.3969/j.issn.1006-5725.2019.16.011

具核梭杆菌感染对结肠癌转化生长因子-β1表达及肿瘤浸润淋巴细胞的影响

卫利民¹, 谷变利², 原翔²

河南科技大学第一附属医院（开元院区） 1甲状腺乳腺肿瘤外科,2肿瘤表观遗传学重点实验室（河南洛阳 471003）

收稿日期:2019-01-01 出版日期:2019-08-27 发布日期:2019-08-27
基金资助:
国家自然科学基金青年科学基金项目（编号：81702820）

Effects of Fusobacterium nucleatum infection on TGF-β1 expression and tumor-infiltrating lymphocytes in colon cancer

WEI Limin^*, GU Bianli, YUAN Xiang.

*Department of General Surgery, the First Affiliated Hospital of Henan University of Science and Technology, Luoyang 471023, China

Received:2019-01-01 Online:2019-08-27 Published:2019-08-27

摘要/Abstract

摘要： 目的探讨具核梭杆菌（Fusobacterium nucleatum,Fn）感染对结肠癌转化生长因子-β1（transforming growth factor-β1,TGF-β1）表达和肿瘤浸润淋巴细胞（tumor infiltrating lymphocytes,TIL）的影响。方法 qPCR和IHC检测结肠癌组织Fn表达和TGF-β1与FoxP3表达;ELISA、qPCR、流式细胞术检测Fn感染对结肠癌细胞TGF-β1表达、TIL、及FoxP3 mRNA的影响。结果 25例癌组织表达Fn,其Fn丰度高于癌旁组织,且TGF-β1与FoxP3表达正相关（P < 0.05）。Fn感染使结肠癌细胞TGF-β1分泌升高及mRNA上调。活化CD4⁺T细胞与Fn感染的HCT116细胞共培养,CD4⁺T细胞增殖较对照组降低,加入TGF-β1抗体结果逆转;初始CD4⁺T细胞与Fn感染的HCT116共培养,FoxP3⁺Treg增加,FoxP3 mRNA上调,加入TGF-β1抗体结果逆转（P < 0.05）。结论具核梭杆菌可能通过上调结肠癌细胞TGF-β1表达,产生局部免疫抑制效应,介导肿瘤细胞免疫逃逸。

关键词: 具核梭杆菌, 结肠癌, TGF-β1, 肿瘤浸润淋巴细胞, 免疫抑制

Abstract: Objective To investigate the effects of Fusobacterium nucleatum （Fn） infection on the expression of TGF-β1 and tumor infiltrating lymphocytes （TIL）s in colon cancer tissues. Methods qPCR and IHC were used to measure the expression of Fn in colon cancer tissues and determine TGF-β1 expression and Foxp3⁺Tregs of cancer tissues. The expression levels of TGF-β1 in 3 types of colon cancer cells were detected by ELISA and qPCR. Flow cytometry （FCM） and qPCR were respectively applied to measure CD4⁺T cell proliferation and FoxP3⁺Tregs and FoxP3 mRNA in the conditions of Fn infection. Results Fn was found in 25 cancer tissues using qPCR. Among the 25 patients who had positive Fn expression, the abundance of Fn in cancer tissues was significantly higher than that in adjacent normal tissues. IHC demonstrated TGF-β1 expression was correlated with FoxP3 expression in cancer tissues infected with Fn （P < 0.05）. TGF-β1 expression excreted from colon cancer cells with Fn infection increased and TGF-β1 mRNA expression was upregulated, when compared with the control groups. FCM analysis showed that the proliferation of activated CD4⁺T cells reduced in the presence of Fn-CM than that of the CM control group when in coculture with Fn-treated HCT116. The inhibitory effect of Fn-CM was partially reversed after adding anti-TGF-β1 neutralizing antibodies, and proliferation rates of activated CD4⁺T cells was increased. Addition of Fn-CM to cultures resulted in a strong generation of Tregs, with CD4⁺CD25⁺FoxP3⁺Treg and FoxP3 mRNA increased compared to the level for the CM control group, whereas neutralization of TGF-β1 in Fn-CM-treated cultures resulted in reversed Results（P < 0.05）. Conclusion Fn infection can inhibit immune response by upregulating the expression of TGF-β1 in colon cancer, which probably facilitated tumor immune escape in colon cancer.

Key words: Fusobacterium nucleatum, colon cancer, TGF-β1, tumor infiltrating lymphocyte, immunosuppression

卫利民, 谷变利, 原翔. 具核梭杆菌感染对结肠癌转化生长因子-β1表达及肿瘤浸润淋巴细胞的影响[J]. 实用医学杂志, 2019, 35(16): 2564-2569.

WEI Limin, GU Bianli, YUAN Xiang.. Effects of Fusobacterium nucleatum infection on TGF-β1 expression and tumor-infiltrating lymphocytes in colon cancer[J]. The Journal of Practical Medicine, 2019, 35(16): 2564-2569.

参考文献 20

[1]	CHEN W, ZHENG R, BAADE P D, et al.Cancer statistics in China, 2015[J]. CA Cancer J Clin, 2016, 66(2): 115-132.
[2]	AMITAY E L, WERNER S, VITAL M, et al.Fusobacterium and colorectal cancer: Causal factor or passenger? Results from a large colorectal cancer screening study[J]. Carcinogenesis, 2017, 38(8):781-788.
[3]	ALLALI I, BOUKHATEM N, BOUGUENOUCH L, et al.Gut microbiome of Moroccan colorectal cancer patients[J]. Med Microbiol Immunol, 2018, 207(3-4):211-225.
[4]	HAN Y W.Commentary: Oral bacteria as drivers for colorectal cancer[J]. J Periodontol, 2014, 85(9):1155-1157.
[5]	GHOLIZADEH P, ESLAMI H, KAFIL H S.Carcinogenesis mechanisms of Fusobacterium nucleatum[J]. Biomed Pharmacother, 2017, 89:918-925.
[6]	MIMA K, SUKAWA Y, NISHIHARA R, et al.Fusobacterium nucleatum and T Cells in colorectal carcinoma[J]. JAMA Oncol, 2015, 1(5):653-661.
[7]	LI Z, ZHANG L J, ZHANG H R, et al.Tumor-derived transforming growth factor-beta is critical for tumor progression and evasion from immune surveillance[J]. Asian Pac J Cancer Prev, 2014, 15(13):5181-5186.
[8]	LI N, XIE C, LU N H.Transforming growth factor-beta: An important mediator in Helicobacter pylori-associated pathogenesis[J]. Front Cell Infect Microbiol, 2015,5:77.
[9]	GIL J H, SEO J W, CHO M S, et al.Role of Treg and TH17 cells of the gastric mucosa in children with Helicobacter pylori gastritis[J]. J Pediatr Gastroenterol Nutr, 2014, 58(2):245-251.
[10]	HAMADA T, ZHANG X, MIMA K, et al.Fusobacterium nucleatum in colorectal cancer relates to immune response differentially by tumor microsatellite instability status[J]. Cancer Immunol Res, 2018, 6(11):1327-1336.
[11]	BAGHERI N, AZADEGAN-DEHKORDI F, RAHIMIAN G, et al.Role of regulatory t-cells in different clinical expressions of Helicobacter pylori infection[J]. Arch Med Res, 2016, 47(4):245-254.
[12]	尹丹, 曹颖, 王礼鑫, 等. 肠型胃癌细胞中酪蛋白激酶2相互作用蛋白1与转化生长因子β1表达的相关性[J]. 实用医学杂志, 2019,35(2):276-280.
[13]	CHEN T, LI Q, ZHANG X, et al.TOX expression decreases with progression of colorectal cancers and is associated with CD4 T-cell density and Fusobacterium nucleatum infection[J]. Hum Pathol, 2018, 79:93-101.
[14]	贾一琼, 朱光发. 调节性B细胞与自身免疫性疾病关系的研究进展[J]. 实用医学杂志, 2018, 34(15):2478-2480.
[15]	METELLI A, SALEM M, WALLACE C H, et al.Immunoregulatory functions and the therapeutic implications of GARP-TGF-β in inflammation and cancer[J]. J Hematol Oncol, 2018, 11(1):24.
[16]	MIYAZONO K, KATSUNO Y, KOINUMA D, et al.Intracellular and extracellular TGF-β signaling in cancer: Some recent topics[J]. Front Med, 2018, 12(4):387-411.
[17]	AHN S H, CHUN S, PARK C, et al.Transcriptome profiling analysis of senescent gingival fibroblasts in response to Fusobacterium nucleatum infection[J]. PLoS One, 2017, 12(11):e0188755.
[18]	LI N, XIE C, LU N H.Transforming growth factor-β: An important mediator in Helicobacter pylori-associated pathogenesis[J]. Front Cell Infect Microbiol, 2015, 4(5):77.
[19]	LEE D W, HAN S W, KANG J K, et al.Association between fusobacterium nucleatum, pathway mutation, and patient prognosis in colorectal cancer[J]. Ann Surg Oncol, 2018, 25(11):3389-3395.
[20]	SAITO T, NISHIKAWA H, WADA H, et al.Two FOXP3(+)CD4(+) T cell subpopulations distinctly control the prognosis of colorectal cancers[J]. Nat Med, 2016, 22(6):679-684.

具核梭杆菌感染对结肠癌转化生长因子-β1表达及肿瘤浸润淋巴细胞的影响

Effects of Fusobacterium nucleatum infection on TGF-β1 expression and tumor-infiltrating lymphocytes in colon cancer

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献 20

相关文章 2

编辑推荐

Metrics

本文评价

[1]	朱朝阳, 赖冬萍, 张涛, 丁明健, 卓少元, 黄晓燕. 慢病毒介导miR⁃222稳定沉默结肠癌细胞系的构建及其生物学作用[J]. 实用医学杂志, 2020, 36(21): 2894-2899.
[2]	钟文德, 邓小燕, 陈广盛. 髓系来源抑制细胞在舌鳞状细胞癌中的作用[J]. 实用医学杂志, 2020, 36(21): 2900-2905.