摘要
目的:探讨趋化因子CCL20/CCR6促进结肠癌SW480细胞侵袭和迁移的分子机制。方法:筛选高表达CCR6的结肠癌SW480细胞,加入外源性重组人CCL20后,采用Transwell、划痕愈合实验检测其侵袭和迁移能力,以免疫荧光、WB实验检测SW480细胞EMT标志蛋白、AKT信号蛋白以及靶标蛋白MMP3的表达;通过MK2206阻断实验验证AKT信号是其作用机制,通过TCGA数据库资源(https://portal.gdc.cancer.gov/)分析CCL20和MMP3在结直肠癌组织中的表达水平及其相关性。结果:趋化因子CCL20能够明显促进结肠癌SW480细胞侵袭和迁移(均P<0.01),其间并不伴随细胞的EMT变化,而是通过AKT信号的激活及下游靶标蛋白MMP3表达上调是其诱因之一;阻断AKT信号能够明显抑制SW480细胞侵袭和迁移能力,且下调MMP3的表达水平(P<0.05或P<0.01)。TCGA平台数据提示,结肠癌组织中CCL20和MMP3的表达明显高于正常肠黏膜组织,且两者呈明显正相关(r=0.051,P<0.01)。结论:趋化因子CCL20通过AKT/MMP3信号轴而非EMT机制促进结肠癌SW480细胞的侵袭和迁移。
Objective: To investigate the molecular mechanism of chemokine CCL20/CCR6 in promoting invasion and migration of colon cancer SW480 cells. Methods: Colorectal cancer SW480 cells with high expression of CCR6 receptor were screened by immunochemistry(IHC). After co-culture with recombinant human CCL20, the invasion and migration of SW480 cells were detected by Transwell assay and Wound-Healing assay, respectively. Expressions of EMT markers, AKT signal protein and target protein MMP3 were detected by immunofluorescence(IF) and WB. AKT signaling pathway as the key mechanism was confirmed by MK2206 blocking assay. The expressions of CCL20 and MMP3 in colorectal cancer tissues as well as their correlation were analyzed by TCGA database resources(https://portal.gdc.cancer.gov/). Results: CCL20 promoted the invasion and migration ability of SW480 cells significantly(all P<0.01), and this was induced by activation of AKT signaling and up-regulation of downstream target protein MMP3, instead of EMT.Blocking AKT signaling could significantly inhibit the invasion and migration of SW480 cells, and down-regulate MMP3 expression(P<0.05). TCGA platform data showed that the expressions of CCL20 and MMP3 in colorectal cancer tissues were significantly higher than those in normal mucosa tissues(P<0.05 or P<0.01), and an evidently positive correlation was found between CCL20 and MMP3(r=0.051, P<0.01). Conclusion: The chemokine CCL20 promotes the invasion and migration of SW480 cells through AKT/MMP3 signal axis, but not the EMT.
作者
程先硕
杨芳
董坚
李云峰
杨之斌
张洪涛
沈焘
刘萍
殷正丰
李强
CHENG Xianshuo;YANG Fang;DONG Jian;LI Yunfeng;YANG Zhibin;ZHANG Hongtao;SHEN Tao;LIU Ping;YIN Zhengfeng;LI Qiang(Colorectal Surgery, the Third Affiliated Hospital of Kunming Medical University, Tumor Hospital of Yunnan Province, Kunming 650118, Yunnan, China;Molecular Oncology Laboratory, Eastern Hepatobiliary Surgery Hospital, Shanghai 200438, China)
出处
《中国肿瘤生物治疗杂志》
CAS
CSCD
北大核心
2019年第6期650-655,共6页
Chinese Journal of Cancer Biotherapy
基金
国家自然科学基金资助项目(No.81560472)
云南省卫生厅-昆明医科大学联合专项[No.2017FE468(-076)]
云南省科技厅重点资助项目(No.2018FA040)
云南省结直肠肿瘤临床重点专科建设经费资助项目(No.2016-0137)~~
