Orai1对鼻咽癌细胞侵袭和上皮-间充质转化的影响及其机制

doi:10.3969/j.issn.1004-616x.2021.06.003

癌变·畸变·突变 ›› 2021, Vol. 33 ›› Issue (6): 420-425.doi: 10.3969/j.issn.1004-616x.2021.06.003

Orai1对鼻咽癌细胞侵袭和上皮-间充质转化的影响及其机制

郭照萌, 张华, 张鹏

深圳市龙岗区耳鼻咽喉医院耳鼻咽喉科/深圳市耳鼻咽喉研究所, 广东深圳 518172

收稿日期:2021-03-04 修回日期:2021-10-08 出版日期:2021-11-30 发布日期:2021-12-04
通讯作者: 张鹏,E-mail:zhangpeng2600@163.com E-mail:zhangpeng2600@163.com
作者简介:郭照萌,E-mail:zhaomengguo1999@163.com。
基金资助:
国家自然科学基金（81700437）；深圳市医学重点学科建设经费资助项目（SZXK039）

Mechanisms of Orai1 on invasion and epithelial-mesenchymal transformation in nasopharyngeal carcinoma cells

GUO Zhaomeng, ZHANG Hua, ZHANG Peng

Department of Otolaryngology, Longgang Ear, Nose and Throat Hospital, Shenzhen Key Laboratory of Ear, Nose and Throat, Shenzhen 518172, Guangdong, China

Received:2021-03-04 Revised:2021-10-08 Online:2021-11-30 Published:2021-12-04

摘要/Abstract

摘要： 目的： 探讨Orai1对鼻咽癌细胞侵袭和上皮-间充质转化的影响及其作用机制。方法： 培养鼻咽上皮细胞NP69和4种鼻咽癌细胞系（CNE1、CNE2、HONE1和5-8F），分别采用实时荧光定量PCR（qPCR）和Western blot法检测细胞中Orai1的mRNA和蛋白表达水平。采用靶向Orai1的短发夹RNA（shRNA）质粒，转染CNE2细胞，设转染对照质粒的CNE2细胞为阴性对照组，钙离子成像检测钙库调节的钙离子内流，Transwell小室检测细胞转移和侵袭，qPCR和Western blot法检测上皮-间充质转化（EMT）标志物E钙黏蛋白（E-cadherin）、N钙黏蛋白（N-cadherin）和波形蛋白（Vimentin）的mRNA和蛋白表达水平。结果： NP69、CNE1、CNE2、HONE1和5-8F细胞中均表达Orai1，且CNE2细胞中表达相对较高，故选择CNE2细胞进行后续试验。与阴性对照组相比，转染Orai1 shRNA后，CNE2细胞中Orai1的mRNA和蛋白水平均显著降低（P<0.05）；CNE2细胞的钙库调节钙离子内流能力、转移/侵袭能力和EMT相关标志蛋白E-cadherin、N-cadherin和Vimentin的mRNA和蛋白表达水平均显著下降（P<0.05）。结论： 抑制Orai1表达可抑制鼻咽癌细胞钙库调节钙离子内流、转移侵袭和EMT进程，提示Orai1可能成为治疗鼻咽癌转移的新靶标和新方向。

关键词: Orai1, 钙离子, 鼻咽癌, 转移, 上皮-间充质转化

Abstract: OBJECTIVE: The study was aimed to investigate mechanisms of Orai1 on nasopharyngeal carcinoma cell invasion and epithelial-mesenchymal transformation (EMT) in vitro. METHODS: A nasopharyngeal epithelial cell line (NP69) and nasopharyngeal carcinoma cell lines (CNE1,CNE2,HONE1 and 5-8F) were cultured. Real-time quantitative PCR and Western blot were used to detect expressions of Orai1 mRNA and protein. The short hairpin RNA (shRNA) plasmid targeting Orai1 was used to down-regulate expressions of Orai1 in CNE2 cells. Store-operated Ca²⁺ entry (SOCE) was detected by calcium imaging,cell migration and invasion were detected by Transwell chamber,and E cadherin,N cadherin and Vimentin were detected by real-time quantitative PCR and Western blot. RESULTS: Orai1 was expressed in NP69,CNE1,CNE2,HONE1 and 5-8F cells,and the expression of Orai1 was higher in CNE2 cells than that in NP69 cells (P<0.05). The mRNA and protein levels of ORAI1 were significantly decreased in CNE2 cells transfected with Orai1 shRNA (P<0.05). SOCE,migration/invasion and EMT were significantly inhibited in Orai1 shRNA transfected CNE2 cells (P<0.05). CONCLUSION: Blockage of ORAI1 expression inhibited SOCE,migration/invasion and EMT process of nasopharyngeal carcinoma cells. Therefore, Orai1 might be a new target and a new direction for the treatment of nasopharyngeal carcinoma metastasis.

Key words: Orai1, Ca²⁺, nasopharyngeal carcinoma, metastasis, epithelial-mesenchymal transformation

中图分类号:

R73-37

郭照萌, 张华, 张鹏. Orai1对鼻咽癌细胞侵袭和上皮-间充质转化的影响及其机制[J]. 癌变·畸变·突变, 2021, 33(6): 420-425.

GUO Zhaomeng, ZHANG Hua, ZHANG Peng. Mechanisms of Orai1 on invasion and epithelial-mesenchymal transformation in nasopharyngeal carcinoma cells[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2021, 33(6): 420-425.

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Orai1对鼻咽癌细胞侵袭和上皮-间充质转化的影响及其机制

Mechanisms of Orai1 on invasion and epithelial-mesenchymal transformation in nasopharyngeal carcinoma cells

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