Liu Wan, Wang Wenjing, Zhang Ning, Di Wen
Department of Obstetrics and Gynecology, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200127, People's Republic of China.
Shanghai Key Laboratory of Gynecologic Oncology, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200127, People's Republic of China.
Onco Targets Ther. 2020 Dec 11;13:12739-12750. doi: 10.2147/OTT.S280309. eCollection 2020.
Chemokine networks play a key and complex role in tumor progression. CCL20 and its unique receptor CCR6 have been reported to mediate malignant biological activities in various cancers, but their role in ovarian cancer metastasis remains unclear.
Our study aims to explore the effect of CCL20-CCR6 axis on ovarian cancer metastasis and its potential mechanism.
The transwell assay was used to detect the cell migration and invasion after CCL20 treatment. The CCK-8 assay was used to detect the cell viability after CCL20 treatment and CCR6 depletion. The mRNA and protein expression were assayed through qRT-PCR and Western blotting. The siRNAs and CRISPR-Cas9 system were adopted to suppress CCR6 expression. Intraperitoneal xenograft mouse model was constructed to test the pro-metastasis effect of CCL20-CCR6 axis in vivo. The differentially expressed genes induced by CCL20 were identified through RNA-sequencing, and immunohistochemistry staining was used to detect their protein expression in tumor tissues.
Our results revealed that CCL20 treatment selectively promoted the migration and invasion of CCR6 ovarian cancer cells, but had no effect on CCR6 cells. Blockade of CCR6 expression effectively reversed the cell migration and invasion induced by CCL20 stimulation. Animal experiment proved that CCL20-CCR6 axis mediated ovarian cancer metastasis in vivo. The differentially expressed genes after CCL20 stimulation were associated with metastasis, and CCL20 induced an increased expression of CDH2 and VCAN and decreased CDH1 expression in cancer cells. Moreover, CCL20 stimulated the expression of N-cadherin and versican in tumor tissues and inhibited the expression of E-cadherin, while CCR6 knockout successfully blocked the expression changes.
Our findings revealed that CCL20-CCR6 axis promotes ovarian cancer metastasis both in vivo and in vitro, probably through increasing cancer cell adhesion and epithelial-mesenchymal transition. Blockade of CCL20-CCR6 axis might become a novel anti-tumor therapeutic target for ovarian cancer.
趋化因子网络在肿瘤进展中起着关键且复杂的作用。据报道，CCL20及其独特受体CCR6在多种癌症中介导恶性生物学活性，但其在卵巢癌转移中的作用仍不清楚。
本研究旨在探讨CCL20-CCR6轴对卵巢癌转移的影响及其潜在机制。
采用Transwell实验检测CCL20处理后细胞的迁移和侵袭能力。采用CCK-8实验检测CCL20处理及CCR6基因敲减后细胞的活力。通过qRT-PCR和蛋白质免疫印迹法检测mRNA和蛋白表达。采用小干扰RNA（siRNAs）和CRISPR-Cas9系统抑制CCR6表达。构建腹腔内异种移植小鼠模型，在体内验证CCL20-CCR6轴的促转移作用。通过RNA测序鉴定CCL20诱导的差异表达基因，并用免疫组织化学染色检测其在肿瘤组织中的蛋白表达。
我们的结果显示，CCL20处理选择性促进CCR6阳性卵巢癌细胞的迁移和侵袭，但对CCR6阴性细胞无影响。阻断CCR6表达可有效逆转CCL20刺激诱导的细胞迁移和侵袭。动物实验证明，CCL20-CCR6轴在体内介导卵巢癌转移。CCL20刺激后的差异表达基因与转移相关，CCL20诱导癌细胞中CDH2和VCAN表达增加，CDH1表达降低。此外，CCL20刺激肿瘤组织中N-钙黏蛋白和多功能蛋白聚糖的表达，抑制E-钙黏蛋白的表达，而CCR6基因敲除成功阻断了这些表达变化。
我们的研究结果表明，CCL20-CCR6轴在体内和体外均促进卵巢癌转移，可能是通过增加癌细胞黏附和上皮-间质转化实现的。阻断CCL20-CCR6轴可能成为卵巢癌新的抗肿瘤治疗靶点。
