低氧条件下巨噬细胞分泌CCL22促进三阴乳腺癌转移

陈敏; 胡绍勇; 何成思; 邹争志

doi:10.6054/j.jscnun.2022030

低氧条件下巨噬细胞分泌CCL22促进三阴乳腺癌转移

doi: 10.6054/j.jscnun.2022030

1.
广州医科大学附属第四医院影像科，广州 511300
2.
南方医科大学第三附属医院影像科，广州 510630
3.
华南师范大学生物光子学研究院激光生命科学研究所暨激光生命科学教育部重点实验室，广州 510631

基金项目:

国家自然科学基金项目 81972479

广东省自然科学基金项目 2019A1515011100

广东省普通高校特色创新类项目 2021KTSCX026

广州市科技计划项目 201904010038

详细信息

通讯作者:
邹争志，Email：zouzhengzhi@m.scnu.edu.cn

中图分类号: R453.9; R34
计量
- 文章访问数: 37
- HTML全文浏览量: 23
- PDF下载量: 25
- 被引次数: 0
出版历程
- 收稿日期: 2021-09-19
- 网络出版日期: 2022-05-12
- 刊出日期: 2022-04-25

Macrophages Secreting CCL22 to Promote Triple-negative Breast Cancer Metastasis Under Hypoxia

1.
Department of Imaging, The Fourth Affiliated Hospital of Guangzhou Medical University, Guangzhou 511300, China
2.
Department of Radiology, The Third Affiliated Hospital of Southern Medical University, Guangzhou 510630, China
3.
MOE Key Laboratory of Laser Life Science & Institute of Laser Life Science, College of Biophotonics, South China Normal University, Guangzhou 510631, China

摘要

摘要: 三阴乳腺癌(Triple Negative Breast Cancer, TNBC)是乳腺癌中恶性程度最高的一种亚型，表现为很高的转移潜能。巨噬细胞，即肿瘤相关巨噬细胞(Tumor-Associated Macrophages, TAM)，在促进TNBC转移中起了重要作用。乳腺癌作为一种实体肿瘤，往往处于缺氧环境中。低氧环境能够促进癌细胞的转移，然而低氧环境中巨噬细胞在促进肿瘤转移中的作用仍然不清楚。在该研究中，THP1细胞被诱导成TAM，经过缺氧培养后，通过Transwell实验检测其促进三阴乳腺癌细胞BT-549和MDA-MB-231的细胞迁移能力；通过尾静脉注射，将MDA-MB-231细胞移植于祼鼠体内，CT扫描，分析了TAM促进TNBC细胞的器官转移能力；通过ELISA实验检测低氧对TAM分泌的肿瘤转移相关因子的影响，通过GDSC在线软件分析了CCL22受体CCR4和其他CCR在乳腺癌组织与正常组织中表达的差异。结果表明低氧条件下巨噬细胞通过分泌CCL22的表达来促进三阴乳腺癌细胞迁移：经过缺氧培养后的TAM显著增强了TNBC细胞迁移能力，以及促进癌细胞在体内向肺转移；低氧诱导TAM分泌CCL22；CCL22受体CCR4在乳腺癌组织中的表达显著高于在正常组织中的。
- 巨噬细胞 /
- 三阴乳腺癌 /
- CCL22 /
- 低氧 /
- CT扫描
Abstract: Triple-negative breast cancer (TNBC) is the most malignant subtype of breast cancer and shows high metastatic potential. Macrophages (or known as tumor-associated macrophages, TAM) play an important role in promoting TNBC metastasis. Breast cancer as a solid tumor often exists in a hypoxic tumor microenvironment. Hypoxia promotes cancer cell metastasis, but the role of macrophages in promoting tumor metastasis in a hypoxic environment is still unclear. THP1 cells were differentiated into TAM and cultured in hypoxia. The migration ability of the TNBC cells BT-549 and MDA-MB-231 induced by TAM was detected through Transwell. MDA-MB-231 was transplanted through tail vein into the nude mice and the organ metastasis ability of TNBC cells induced by TAM was analyzed through CT scan. The effect of hypoxia on the secretion of several factors related to tumor metastasis in TAM was detected with the ELISA test. The difference between CCL22 receptor CCR4 and other CCR expression in breast cancer tissues and normal tissues was analyzed with the GDSC online software. The results showed that hypoxia-induced TAM significantly enhanced the migration of TNBC cells. Hypoxia induced CCL22 expression in TAM. Moreover, CCR4 expression in breast cancer tissues was significantly higher than that in normal tissues. In conclusion, TAM could promote the migration of TNBC cells through secretion of CCL22 under hypoxia.
- macrophages /
- triple-negative breast cancer /
- CCL22 /
- hypoxia /
- CT scan

HTML全文

图 1 3种CM处理的三阴乳腺癌细胞Transwell实验

注：M0代表M0巨噬细胞，TAM代表肿瘤相关巨噬细胞，Hyp代表低氧(hypoxia)；数据表示为x±s；* * *表示P＜0.001。

Figure 1. The Transwell-detected cell migration of TNBC treated with three CM

下载: 全尺寸图片幻灯片

图 2 RT-PCR检测3种CM处理的MDA-MB-231细胞迁移相关基因表达

注：M0代表M0巨噬细胞，TAM代表肿瘤相关巨噬细胞，Hyp代表低氧(hypoxia)；数据表示为x±s；* * *表示P＜0.001。

Figure 2. The RT-PCR-detected expression of migration-associated gene in MDA-MB-231 cells treated with three CM

下载: 全尺寸图片幻灯片

图 3 CT扫描检测三阴乳腺癌细胞肺转移灶

注：红色箭头代表转移灶；TAM代表肿瘤相关巨噬细胞，Hyp代表低氧(hypoxia)；数据表示为x±s；* *表示P＜0.01，n=5。

Figure 3. The CT-detected TNBC lung metastasis

下载: 全尺寸图片幻灯片

图 4 通过ELISA方法检测巨噬细胞分泌细胞因子的质量浓度

注: TAM代表肿瘤相关巨噬细胞，Hyp代表低氧(hypoxia)；数据表示为x±s；* * *表示P＜0.001。

Figure 4. The ELISA-detected concentrations of cytokines secreted by macrophage

下载: 全尺寸图片幻灯片

图 5 不同质量浓度的CCL22诱导的三阴乳腺癌细胞Transwell实验

注: 数据表示为x±s；*表示P＜0.05，* * *表示P＜0.001。

Figure 5. The Transwell-detected TNBC cells migration induced by CCL22 of different concentrations

下载: 全尺寸图片幻灯片

表 1 CCR家族基因在乳腺癌组织与正常组织中表达的差异与比值

Table 1. The expression and ratio of CCR family members in breast cancer tissue and normal tissue

基因	正常组织表达值(N)	肿瘤组织表达值(T)	T/N	P值
CCR1	270	320	1.19	0.19
CCR2	98.00	110	1.12	0.32
CCR3	1.90	3.50	1.84	7.80×10^-3
CCR4	15.00	49.00	3.27	2.20×10^-7
CCR5	140	270	1.93	3.00×10^-5

下载: 导出CSV

参考文献(19)

[1]	赖丽梨, 靳焕, 段华英, 等. 巨噬细胞增强宫颈癌细胞对SN－38的抗性[J]. 华南师范大学学报(自然科学版), 2021, 53(1): 63-69. doi: 10.6054/j.jscnun.2021010 LAI L L, JIN H, DUAN H Y, et al. Macrophage's promotion of cervical cancer cell resistance to SN-38[J]. Journal of South China Normal University(Natural Science Edition), 2021, 53(1): 63-69. doi: 10.6054/j.jscnun.2021010
[2]	关燕清, 杨鑫华, 何丽梅, 等. TNF与IFN协同诱导癌细胞凋亡机制的研究进展[J]. 华南师范大学学报(自然科学版), 2006, 8(3): 122-126. doi: 10.3969/j.issn.1000-5463.2006.03.022 GUAN Y Q, YANG X H, HE L M, et al. Research progress of synergic mechanisms of apoptosis induced by TNF and IFN[J]. Journal of South China Normal University(Natural Science Edition), 2006, 8(3): 122-126. doi: 10.3969/j.issn.1000-5463.2006.03.022
[3]	CHEN J Q, YAO Y D, CHANG G, et al. CCL18 from tum-or-associated macrophages promotes breast cancer metastasis via PITPNM3[J]. Cancer Cell, 2011, 19(6): 814-816. doi: 10.1016/j.ccr.2011.05.024
[4]	SU S C, LIU Q, CHEN J Q, et al. A positive feedback loop between mesenchymal-like cancer cells and macrophages is essential to breast cancer metastasis[J]. Cancer Cell, 2014, 25(5): 605-620. doi: 10.1016/j.ccr.2014.03.021
[5]	MONTAGNER M, ENZO E, FORCATO M, et al. SHARP1 suppresses breast cancer metastasis by promoting degradation of hypoxia-inducible factors[J]. Nature, 2012, 487: 380-384. doi: 10.1038/nature11207
[6]	KORBECKI J, KOJDER K, SIMIŃSKA D, et al. CC chemokines in a tumor: a review of pro-cancer and anti-cancer properties of the ligands of receptors CCR1, CCR2, CCR3, and CCR4[J]. International Journal of Molecular Sciences, 2020, 21(21): 8412-8440. doi: 10.3390/ijms21218412
[7]	TANG J M, ZHONG G S, ZHANG H B, et al. LncRNA DANCR upregulates PI3K/AKT signaling through activating serine phosphorylation of RXRA[J]. Cell Death & Disease, 2018, 9(12): 1167-1178.
[8]	SHOME R, GHOSH S S. Tweaking EMT and MDR dynamics to constrain triple-negative breast cancer invasiveness by EGFR and Wnt/β-catenin signaling regulation[J]. Cellular Oncology, 2021, 44(2): 405-422. doi: 10.1007/s13402-020-00576-8
[9]	SAXENA K, JOLLY M K, BALAMURUGAN K. Hypoxia, partial EMT and collective migration: emerging culprits in metastasis[J]. Translational Oncology, 2020, 13(11): 100845-100858. doi: 10.1016/j.tranon.2020.100845
[10]	BRANCO-PRICE C, ZHANG N, SCHNELLE M, et al. Endothelial cell HIF-1α and HIF-2α differentially regulate metastatic success[J]. Cancer Cell, 2012, 21(1): 52-65. doi: 10.1016/j.ccr.2011.11.017
[11]	CHEN Y B, SONG Y C, DU W, et al. Tumor-associated macrophages: an accomplice in solid tumor progression[J]. Journal of Biomedical Science, 2019, 26(1): 1-13. doi: 10.1186/s12929-018-0495-4
[12]	邹争志, 聂培培, 倪艺榕, 等. 硫链丝菌素诱导非小细胞肺癌细胞自噬性死亡[J]. 激光生物学报, 2014, 23(1): 33-37. https://www.cnki.com.cn/Article/CJFDTOTAL-JGSW201401007.htm ZOU Z Z, NIE P P, NI Y R, et al. Thiostrepton induces autophagic cell death in non-small-cell lung cancer cells[J]. Acta Laser Biology Sinica, 2014, 23(1): 33-37. https://www.cnki.com.cn/Article/CJFDTOTAL-JGSW201401007.htm
[13]	TU D Y, DOU J, WANG M K, et al. M2 macrophages contribute to cell proliferation and migration of breast cancer[J]. Cell Biology International, 2020, 45(4): 831-838.
[14]	ALLEN S G, CHEN Y C, MADDEN J M, et al. Macrophages enhance migration in inflammatory breast cancer cells via RhoC GTPase signaling[J]. Scientific Reports, 2016, 6(1): 39190-39200. doi: 10.1038/srep39190
[15]	SEMENZA G L. Molecular mechanisms mediating metastasis of hypoxic breast cancer cells[J]. Trends in Molecular Medicine, 2012, 18(9): 534-543.
[16]	QIAO J H, CHEN Y B, MI Y J, et al. Macrophages confer resistance to BET inhibition in triple-negative breast cancer by upregulating IKBKE[J]. Biochemical Pharmacology, 2020, 180: 114-126.
[17]	ARAUJO-PIRES A C, VIEIRA A E, FRANCISCONI C F, et al. IL-4/CCL22/CCR4 axis controls regulatory T-cell migration that suppresses inflammatory bone loss in murine experimental periodontitis[J]. Journal of Bone and Mineral Research, 2015, 30(3): 412-422.
[18]	YANG X X, BAO M C, FANG Y, et al. STAT3/HIF-1α signaling activation mediates peritoneal fibrosis induced by high glucose[J]. Journal of Translational Medicine, 2021, 19(1): 283-297.
[19]	NICOLAY J P, ALBRECHT J D, ALBERTI-VIOLETTI S, et al. CCR4 in cutaneous T-cell lymphoma: therapeutic targeting of a pathogenic driver[J]. European Journal of Immunology, 2021, 51(7): 1660-1671.