DCLK1敲除对食管鳞癌细胞迁移和侵袭能力的影响

doi:10.3969/j.issn.1006-7795.2019.03.017

首都医科大学学报 ›› 2019, Vol. 40 ›› Issue (3): 417-423.doi: 10.3969/j.issn.1006-7795.2019.03.017

DCLK1敲除对食管鳞癌细胞迁移和侵袭能力的影响

范晓娜¹, 姚健楠², 葛洋², 安广宇², 李鹰¹

1. 首都医科大学附属北京朝阳医院医学研究中心, 北京 100020;
2. 首都医科大学附属北京朝阳医院肿瘤科, 北京 100020

收稿日期:2019-03-12 出版日期:2019-05-21 发布日期:2019-06-13
通讯作者: 李鹰 E-mail:leeeying2013@hotmail.com
基金资助:
国家自然科学基金（81802738）。

Effect of DCLK1 knockout on migration and invasion of esophageal squamous carcinoma cells

Fan Xiaona¹, Yao Jiannan², Ge Yang², An Guangyu², Li Ying¹

1. Medical Research Center, Beijing Chaoyang Hospital, Capital Medical University, Beijing 100020, China;
2. Department of Oncology, Beijing Chaoyang Hospital, Capital Medical University, Beijing 100020, China

Received:2019-03-12 Online:2019-05-21 Published:2019-06-13
Supported by:
This study was supported by National Natural Science Foundation of China (81802738).

摘要/Abstract

摘要： 目的研究双皮质素样激酶1（doublecortin-like kinase 1，DCLK1）的敲除对食管鳞癌细胞迁移和侵袭能力的影响及其潜在的作用机制。方法使用CRISPR/Cas9基因编辑技术靶向敲除食管鳞癌细胞（Kyse450，Kyse70）的DCLK1基因。通过细胞划痕实验和Transwell实验评估DCLK1敲除对食管鳞癌细胞迁移和侵袭能力的影响。Western blotting法检测DCLK1敲除细胞系中上皮-间质转化（epithelial-mesenchymal transition，EMT）相关转录因子的表达变化。通过癌症基因组图谱（The Cancer Genome Atla，TCGA）数据库进一步分析食管鳞癌患者（n=95）中DCLK1表达和EMT相关转录因子之间的相关性。结果 CRISPR/Cas9基因编辑技术能够有效地敲除食管鳞癌细胞系DCLK1基因并影响其表达。DCLK1缺失显著抑制食管鳞癌细胞的体外迁移和侵袭能力（P<0.05），并上调上皮标志物E-cadherin的表达，下调间质标志物如Vimentin、ZEB1、Slug和Snail的表达（P<0.05）。另外TCGA数据库分析显示食管鳞癌患者中DCLK1与EMT相关转录因子存在相关性。结论 DCLK1敲除可通过抑制EMT进程影响食管鳞癌细胞的迁移和侵袭。

关键词: 双皮质素样激酶1, 食管鳞癌, 上皮间质转化, 迁移, 侵袭

Abstract: Objective To investigate the effect of doublecortin-like kinase 1 (DCLK1) knockout on the migration and invasion of esophageal squamous carcinoma (ESCC) cells and its potential mechanism. Methods CRISPR/Cas9 genome-editing technique was used to knock out DCLK1 gene in ESCC cells (Kyse450, Kyse70). Wound healing assays and Transwell assays were performed to assess the effects of DCLK1 knockout on migration and invasion of ESCC cells. Western blotting was used to detect the expression of epithelial-mesenchymal transition (EMT) related transcription factors in DCLK1 knockout cell lines. The correlation between DCLK1 expression and epithelial-mesenchymal transition (EMT) associated transcription factors were analyzed among 95 ESCC patients from The Cancer Genome Atla (TCGA) database.Results CRISPR/Cas9 technology efficiently disrupted the DCLK1 gene and abrogated its expression in ESCC cell lines. DCLK1 deficiency significantly inhibited ESCC cells migration and invasion (P<0.05), and up-regulated the expression of the epithelial marker E-cadherin, but decreased the levels of mesenchymal markers, such as Vimentin, ZEB1, Slug and Snail in ESCC cell lines (P<0.05). In addition, TCGA database analysis showed that DCLK1 is associated with EMT associated transcription factors in patients with ESCC. Conclusion DCLK1 knockout can affect the migration and invasion of ESCC cells by inhibiting EMT process.

Key words: doublecortin-like kinase 1(DCLK1), esophageal squamous cell carcinoma, epithelial-mesenchymal transition, migration, invasion

中图分类号:

R735.1

范晓娜, 姚健楠, 葛洋, 安广宇, 李鹰. DCLK1敲除对食管鳞癌细胞迁移和侵袭能力的影响[J]. 首都医科大学学报, 2019, 40(3): 417-423.

Fan Xiaona, Yao Jiannan, Ge Yang, An Guangyu, Li Ying. Effect of DCLK1 knockout on migration and invasion of esophageal squamous carcinoma cells[J]. Journal of Capital Medical University, 2019, 40(3): 417-423.

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DCLK1敲除对食管鳞癌细胞迁移和侵袭能力的影响

Effect of DCLK1 knockout on migration and invasion of esophageal squamous carcinoma cells

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