华蟾素注射液对人食管癌细胞增殖、侵袭和化疗敏感性的影响及其机制

doi:10.3969/j.issn.1004-616x.2017.02.004

癌变·畸变·突变 ›› 2017, Vol. 29 ›› Issue (2): 96-101.doi: 10.3969/j.issn.1004-616x.2017.02.004

华蟾素注射液对人食管癌细胞增殖、侵袭和化疗敏感性的影响及其机制

杨兴肖¹, 马鸣², 单保恩³, 李幼梅⁴, 宋姮⁴, 刘志坤⁴, 祝淑钗⁴

1. 河北医科大学第四医院感染管理科, 河北石家庄 050011;
2. 河北医科大学第四医院检验科, 河北石家庄 050011;
3. 河北医科大学第四医院科研中心, 河北石家庄 050011;
4. 河北医科大学第四医院放疗科, 河北石家庄 050011

收稿日期:2016-08-24 修回日期:2016-12-12 出版日期:2017-03-31 发布日期:2017-03-31
通讯作者: 单保恩,E-mail:shanbaoen@163.com E-mail:shanbaoen@163.com
作者简介:杨兴肖,E-mail:yangxingxiao2007@sina.com。
基金资助:
国家自然科学基金项目（81372416）；河北省中医药管理局项目（2015131）；河北省医学科学研究所项目（20160183，20160171）

Effect of cinobufacini on chemotherapy sensitivity and toxicity of human esophageal carcinoma cells in vitro

YANG Xingxiao¹, MA Ming², SHAN Baoen³, LI Youmei⁴, SONG Heng⁴, LIU Zhikun⁴, ZHU Shuchai⁴

1. Department of Infection Management, The Fourth Affiliated Hospital of Hebei Medical University, Shijiazhuang 050011;
2. Department of Laboratory, The Fourth Affiliated Hospital of Hebei Medical University, Shijiazhuang 050011;
3. Research Centre, The Fourth Affiliated Hospital of Hebei Medical University, Shijiazhuang 050011;
4. Department of Radiation Oncology, The Fourth Affiliated Hospital of Hebei Medical University, Shijiazhuang 050011, Hebei, China

Received:2016-08-24 Revised:2016-12-12 Online:2017-03-31 Published:2017-03-31

摘要/Abstract

摘要：

目的：研究华蟾素注射液对食管癌细胞增殖、侵袭及化疗敏感性的调控作用，并初步探讨其作用机制。方法：应用四甲基噻唑蓝（MTT）法检测华蟾素注射液对人食管癌ECA109细胞增殖及化疗敏感性的影响；流式细胞术检测华蟾素注射液联合化疗药物（顺铂或表柔比星）对ECA109细胞凋亡水平的影响；Transwell小室实验检测华蟾素注射液对ECA109细胞迁移及侵袭能力的影响；Western blot技术检测华蟾素注射液对ECA109细胞中与侵袭、上皮-间质转化（EMT）及细胞耐药相关蛋白（Snail、Twist、MMP2、Vimentin、P-gp及E-cadherin）表达水平的影响。结果：与空白对照组比较，华蟾素注射液可显著抑制食管癌ECA109细胞的增殖能力，且呈时间和剂量效应关系（时间效应，r=0.985，P=0.000；剂量效应，r=0.988，P=0.000），并可明显促进该细胞的化疗敏感性（P=0.000）；华蟾素注射液可明显抑制ECA109细胞侵袭和迁移能力（P均=0.000）；华蟾素注射液可明显抑制Snail、Twist、MMP2、Vimentin及P-gp蛋白的表达水平，同时促进细胞表达E-cadherin蛋白。结论：华蟾素注射液能有效抑制人食管癌ECA109细胞的增殖及侵袭活性，并可有效增强该细胞对化疗药物的敏感性，其机制可能与该药抑制肿瘤细胞EMT过程有关。

关键词: 华蟾素, 化疗敏感性, 上皮-间质转化, 增殖, 侵袭

Abstract:

OBJECTIVE: To investigate the effects of cinobufacini on proliferation,invasion and chemotherapy sensitivity of human esophageal carcinoma,and to analyze its impact on epithelial-mesenchymal transition and its mechanism. METHODS: The effects of cinobufacini on proliferation and chemotherapy sensitivity of human esophagus cancer ECA109 cells were measured by the methyl thiazolyl tetrazolium (MTT) assay. After treatment with cinobufacini,cell apoptosis was analyzed and chemotherapy sensitivity by flow cytometry (FCW). In addition,transwell assay was employed to examine viability,migration and invasion. The effects of cinobufacini on invasion,epithelial-mesenchymal transition of ECA109 and multidrug resistance (MDR)-related proteins were examined by Western blot assay. RESULTS: Cinobufacini treatment significantly inhibited ECA109 cell proliferation in both time- and dose-dependent ways (r=0.985,P=0.000;r=0.988,P=0.000),and enhanced chemotherapy sensitivity compared to that in the control group (P=0.000). Cell migration and invasion were significantly suppressed (both P=0.000). In addition,the treatment also down-regulated the expression levels of Snail,Twist,MMP2,Vimentin and P-gp,but up-regulated the expression of E-cadherin. CONCLUSION: Cinobufacini effectively inhibited the proliferation and invasion of human esophageal carcinoma ECA109 cell,and enhance its chemotherapy sensitivity in culture. The mechanism might be partly related to the inhibition epithelial-mesenchymal transition by cinobufacini.

Key words: cinobufacini, chemotherapy sensitivity, epithelial-mesenchymal transition, proliferation, invasion

中图分类号:

R730.5

杨兴肖, 马鸣, 单保恩, 李幼梅, 宋姮, 刘志坤, 祝淑钗. 华蟾素注射液对人食管癌细胞增殖、侵袭和化疗敏感性的影响及其机制[J]. 癌变·畸变·突变, 2017, 29(2): 96-101.

YANG Xingxiao, MA Ming, SHAN Baoen, LI Youmei, SONG Heng, LIU Zhikun, ZHU Shuchai. Effect of cinobufacini on chemotherapy sensitivity and toxicity of human esophageal carcinoma cells in vitro[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2017, 29(2): 96-101.

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华蟾素注射液对人食管癌细胞增殖、侵袭和化疗敏感性的影响及其机制

Effect of cinobufacini on chemotherapy sensitivity and toxicity of human esophageal carcinoma cells in vitro

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