解剖学报 ›› 2019, Vol. 50 ›› Issue (5): 601-607.doi: 10.16098/j.issn.0529-1356.2019.05.010

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冬凌草甲素对人食管鳞癌细胞系KYSE-150和KYSE-450增殖及迁移的影响

黄坷坷1,2,3 刘玉珍1,2,3 陈彦民1 陈智1 刘丹辉1 赵宝生1,2*   

  1. 1.新乡医学院第一附属医院胸外科,河南 卫辉 53100; 2.新乡医学院食管癌研究所,河南 卫辉 453100; 3.新乡医学院第一附属医院河南省神经病学研究所,河南 卫辉 453100
  • 收稿日期:2018-08-15 修回日期:2018-10-26 出版日期:2019-10-06 发布日期:2019-12-10
  • 通讯作者: 赵宝生 E-mail:drbszhao@xxmu.edu.cn
  • 基金资助:
    miR-212和人食管癌转移的相关性及其作用机制的研究;新乡医学院第一附属医院博士基金

 Inhibitory effect of oridonin on proliferation and migration of human esophageal squamous cancer cell lines KYSE-150 and KYSE-450

HUANG Ke-ke 1,2,3 LIU Yu-zhen 1,2,3 CHEN Yan-min1 CHEN Zhi1 LIU Dan-hui1 ZHAO Bao-sheng 1,2*   

  1. 1.Department of Thoracic Surgery, the First Affiliated Hospital of Xinxiang Medical University, He’nan Weihui 453100, China; 2.Esophageal Cancer Institute of Xinxiang Medical University, He’nan Weihui 453100, China; 3.He’nan Neurology Institute at the First Affiliated Hospital of Xinxiang Medical University, He’nan Weihui 453100, China
  • Received:2018-08-15 Revised:2018-10-26 Online:2019-10-06 Published:2019-12-10
  • Contact: ZHAO Bao-sheng E-mail:drbszhao@xxmu.edu.cn

摘要:

目的 探讨冬凌草甲素(ORI)对食管鳞癌细胞系KYSE-150和KYSE-450增殖、凋亡、周期及迁移的作用。 方法 MTT法检测ORI对食管癌细胞增殖的影响;集落形成实验检测ORI对集落形成的影响;流式细胞术检测ORI对食管癌细胞凋亡和周期的影响;Transwell迁移实验检测ORI对食管癌细胞迁移的作用;Western blotting检测ORI对抗凋亡蛋白Bcl-2、细胞周期抑制蛋白p21Cip1/Waf1及上皮-间质转化(EMT)标志蛋白表达水平的影响。 结果 ORI对KYSE-150和KYSE-450细胞的增殖、迁移和集落形成有显著的抑制作用(P<0.05),且抑制作用呈一定的时间、剂量依赖性;流式结果显示,随着ORI浓度的增加,细胞的凋亡率明显增加(P<0.05),G2/M期细胞比例显著增加(P<0.05),G0/G1期细胞比例明显下降(P<0.05);ORI处理食管癌细胞48 h,Bcl-2、间质细胞标志蛋白,波形蛋白(vimentin)、β-连环蛋白(β-catenin)表达下调,p21Cip1/Waf1、上皮细胞标志蛋白,E-钙黏蛋白(E-cadherin)表达上调。 结论 冬凌草甲素可能通过诱导细胞凋亡,阻滞细胞在G2/M期抑制食管癌细胞的增殖,并通过抑制EMT转化从而抑制食管癌细胞的迁移。

关键词: 食管鳞癌细胞, 冬凌草甲素, 增殖, 迁移, 流式细胞术,

Abstract:

Objective To explore the effect of oridonin (ORI) on proliferation, apoptosis, cell cycle and migration of esophageal squamous carcinoma cell (ESCC) lines KYSE-150 and KYSE-450. Methods The effect of ORI on the proliferation and clony formation of esophageal cancer cells were detected by MTT and colony formation assays. Flow cytometry was performed to examine the impact of ORI on cell apoptosis and cell cycle. Transwell assay was applied to detect the role of ORI on cell migration. The effect of ORI on the expression of anti-apoptotic protein Bcl-2, cell cycle inhibitory protein p21Cip1/Waf1, epithelialmesenchymal transition (EMT) related markers were examined by Western blotting. Results ORI had a significant inhibitory effect on the proliferation, migration and clone formation of KYSE-150 and KYSE-450 cells (P<0.05) in a time and dose-dependent manner. With the increase of ORI concentration, apoptosis rate and the proportion of cells in G2/M phase increased significantly (P<0.05), and the proportion of cells in G0/G1 phase decreased significantly (P<0.05). Bcl-2, vimentin and β-catenin were down-regulated and p21Cip1/Waf1, E-cadherin were up-regulated after treatment of ORI on ESCC cells for 48 hours. Conclusion ORI may inhibit ESCC cell proliferation and clony formation by inducing apoptosis and resting cells in G2/M phase, and suppress ESCC cell migration via inhibiting EMT process.

Key words: Esophageal squamous carcinoma cell, Oridonin, Proliferation, Migration, Flow cytometry, Human