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雷公藤红素抑制鼻咽癌细胞的增殖、迁移及上皮间质转化
张雪1△,张登海1△,陈晓平2,滕伟强3,彭彬1,张燚2,王莹1,曹帆帆1,薛晓成2*
0
(1. 海军军医大学(第二军医大学)附属公利医院中心实验室, 上海 200135;
2. 海军军医大学(第二军医大学)附属公利医院耳鼻咽喉科, 上海 200135;
3. 海军军医大学(第二军医大学)长海医院耳鼻咽喉头颈外科, 上海 200433
共同第一作者
*通信作者)
摘要:
目的 研究雷公藤红素在鼻咽癌细胞增殖、迁移及上皮间质转化中的作用。方法 使用终浓度为1.8 μmol/L的雷公藤红素处理人鼻咽癌HNE1、CNE2细胞,并设二甲基亚砜(DMSO)对照组。用CCK-8法检测HNE1、CNE2细胞的增殖能力,细胞划痕实验检测细胞迁移能力,细胞克隆形成实验检测细胞的克隆形成数,细胞黏附与分离实验检测细胞的黏附、分离能力,蛋白质印迹法检测细胞上皮间质转化蛋白(E-钙黏蛋白、β-连环蛋白、N-钙黏蛋白、波形蛋白)的表达。结果 与DMSO对照组比较,采用1.8 μmol/L雷公藤红素处理24、48、72 h后,HNE1细胞的增殖能力均降低(P均<0.05),处理48、72 h后,CNE2细胞的增殖能力均降低(P均<0.05);处理48 h后,HNE1、CNE2细胞的迁移能力均下降(P均<0.05);处理2周后,HNE1、CNE2细胞的克隆形成数均减少(P均<0.05);处理1 h后,HNE1、CNE2细胞的黏附率降低(P<0.05);处理24 h后,HNE1、CNE2细胞的分离率降低(P<0.05);处理6 h后,HNE1、CNE2细胞中E-钙黏蛋白、β-连环蛋白的表达升高(P<0.05),N-钙黏蛋白、波形蛋白的表达降低(P均<0.05)。结论 雷公藤红素可抑制鼻咽癌HNE1、CNE2细胞的增殖、迁移及上皮间质转化。
关键词:  鼻咽肿瘤  雷公藤红素  上皮间质转化  细胞增殖  细胞迁移
DOI:10.16781/j.0258-879x.2018.06.0640
投稿时间:2018-01-27修订日期:2018-06-01
基金项目:上海市浦东新区卫生系统重点学科群(PWZxq2017-04),上海市浦东新区公利医院青年基金(2015YQNJJ-01),上海市浦东新区公利医院青年英才项目(GLRq2017-01,GLRq2017-05).
Celastrol inhibits proliferation, migration and epithelial-mesenchymal transition of nasopharyngeal carcinoma cells
ZHANG Xue1△,ZHANG Deng-hai1△,CHEN Xiao-ping2,TENG Wei-qiang3,PENG Bin1,ZHANG Yi2,WANG Ying1,CAO Fan-fan1,XUE Xiao-cheng2*
(1. Central Laboratory, Gongli Hospital, Navy Medical University(Second Military Medical University), Shanghai 200135, China;
2. Department of Otolaryngology, Gongli Hospital, Navy Medical University(Second Military Medical University), Shanghai 200135, China;
3. Department of Otolaryngology Head and Neck Surgery, Changhai Hospital, Navy Medical University(Second Military Medical University), Shanghai 200433, China
Co-first authors.
* Corresponding author)
Abstract:
Objective To investigate the effect of celastrol on proliferation, migration and epithelial-mesenchymal transition of nasopharyngeal carcinoma cells. Methods Human nasopharyngeal carcinoma cells HNE1 and CNE2 were treated with 1.8 μmol/L celastrol, and the cells treated with equal volume of dimethyl sulfoxide (DMSO) were used as control. The proliferation of HNE1 and CNE2 cells was detected by CCK-8 assay, the migration of cells was detected by cell scratch test, the clone formation was detected by cell colony formation assay, the adhesion and separation abilities of cells were examined by cell adhesion and separation experiments, and the expression of E-cadherin, β-catenin, N-cadherin and vimentin was detected by Western blotting analysis. Results Compared with the DMSO group, the proliferation of HNE1 cells was significantly decreased after treated with 1.8 μmol/L celastrol for 24, 48 and 72 h (all P<0.05), and the proliferation of CNE2 cells was significantly decreased for 48 and 72 h (both P<0.05); the migration ability of HNE1 and CNE2 cells was significantly decreased after treated with 1.8 μmol/L celastrol for 48 h (both P<0.05); and the clone formation of HNE1 and CNE2 cells was significantly decreased after treated with 1.8 μmol/L celastrol for two weeks (both P<0.05). The adhesion rate of HNE1 and CNE2 cells 1 h after treatment and separation rate 24 h after treatment in the celastrol groups were significantly lower than those in the corresponding DMSO group (both P<0.05). Western blotting analysis showed that the expressions of E-cadherin and β-catenin in HNE1 and CNE2 cells in the celastrol groups were significantly increased compared with the DMSO group, while the expressions of N-cadherin and vimentin were significantly decreased 6 h after treatment (all P<0.05). Conclusion Celastrol can inhibit the proliferation, migration and epithelial-mesenchymal transition of nasopharyngeal carcinoma cells HNE1 and CNE2.
Key words:  nasopharyngeal neoplasms  celastrol  epithelial-mesenchymal transition  cell proliferation  cell migration