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CTCF沉默的miR-137促进食管鳞状细胞癌的上皮-间质转化和放射抗性。

CTCF-silenced miR-137 contributes to EMT and radioresistance in esophageal squamous cell carcinoma.

作者信息

Xu Shuwen, Li Xiaofeng, Li Longfei, Wang Yufeng, Geng Chong, Guo Feng, Zhang Tao, Du Aonan, Lu Zhiwei, Hui Hua, Wang Qiang

机构信息

Department of Head and Neck Surgery, Xuzhou Cancer Hospital, Xuzhou, 221005, Jiangsu, China.

Department of Radiology, Xuzhou Cancer Hospital, Xuzhou, 221005, Jiangsu, China.

出版信息

Cancer Cell Int. 2021 Mar 8;21(1):155. doi: 10.1186/s12935-020-01740-8.

DOI:10.1186/s12935-020-01740-8
PMID:33685449
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7938596/
Abstract

BACKGROUND

Esophageal squamous cell carcinoma (ESCC) is one of the most malignant tumors in gastrointestinal system. MicroRNAs (miRNAs) have been reported to be implicated in cancer development. However, the role of miR-137 has not been fully revealed in ESCC.

METHODS

Quantitative real-time polymerase chain reaction (qRT-PCR) and western blot analyses were separately used to examine RNA level and protein level. 5-ethynyl-2'-deoxyuridine (EdU) assay, transwell assays and flow cytometry analyses were conducted to assess biological behaviors of ESCC cells. Additionally, the interaction between genes were analyzed via Chromatin Immunoprecipitation (ChIP) assay, RNA Binding Protein Immunoprecipitation (RIP) assay, RNA pull down assay and luciferase reporter assay.

RESULTS

MiR-137 was down-regulated in ESCC cells. Upregulation of miR-137 hindered ESCC cell proliferation, migration, invasion and epithelial mesenchymal transition (EMT). Besides, miR-137 enhanced the sensitivity of ESCC cells to irradiation. Moreover, CCCTC-binding factor (CTCF) inactivated miR-137 transcription in ESCC cells. Furthermore, we revealed enhancer of zeste 2 polycomb repressive complex 2 subunit (EZH2) and paxillin (PXN) as the downstream targets of miR-137. In turn, EZH2 was recruited by CTCF and induced methylation in miR-137 promoter.

CONCLUSION

CTCF/Suz12/EZH2 complex-silenced miR-137 facilitates ESCC progression and radioresistance by targeting EZH2 and PXN.

摘要

背景

食管鳞状细胞癌（ESCC）是胃肠道系统中最具侵袭性的肿瘤之一。据报道，微小RNA（miRNA）与癌症发展有关。然而，miR-137在ESCC中的作用尚未完全明确。

方法

分别采用定量实时聚合酶链反应（qRT-PCR）和蛋白质免疫印迹法检测RNA水平和蛋白质水平。采用5-乙炔基-2'-脱氧尿苷（EdU）检测法、Transwell检测法和流式细胞术分析评估ESCC细胞的生物学行为。此外，通过染色质免疫沉淀（ChIP）检测法、RNA结合蛋白免疫沉淀（RIP）检测法、RNA下拉检测法和荧光素酶报告基因检测法分析基因之间的相互作用。

结果

ESCC细胞中miR-137表达下调。上调miR-137可抑制ESCC细胞的增殖、迁移、侵袭及上皮-间质转化（EMT）。此外，miR-137增强了ESCC细胞对放疗的敏感性。此外，CCCTC结合因子（CTCF）使ESCC细胞中miR-137转录失活。此外，我们发现zeste基因增强子同源物2多梳抑制复合物2亚基（EZH2）和桩蛋白（PXN）是miR-137的下游靶点。反过来，EZH2被CTCF招募并诱导miR-137启动子甲基化。

结论

CTCF/Suz12/EZH2复合物沉默的miR-137通过靶向EZH2和PXN促进ESCC进展和放疗抵抗。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bd16/7938596/fa3d42974220/12935_2020_1740_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bd16/7938596/d8ad032ed9f0/12935_2020_1740_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bd16/7938596/50b2c1059319/12935_2020_1740_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bd16/7938596/fc668e97efc8/12935_2020_1740_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bd16/7938596/96644d637084/12935_2020_1740_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bd16/7938596/fa3d42974220/12935_2020_1740_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bd16/7938596/d8ad032ed9f0/12935_2020_1740_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bd16/7938596/50b2c1059319/12935_2020_1740_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bd16/7938596/fc668e97efc8/12935_2020_1740_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bd16/7938596/96644d637084/12935_2020_1740_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bd16/7938596/fa3d42974220/12935_2020_1740_Fig5_HTML.jpg

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