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ErbB3结合蛋白1的上调抑制食管癌细胞生长

姜浩, 刘冬平, 谢东, 魏东芝

华东理工大学生物反应器工程国家重点实验室，上海 200237；中国科学院上海生命科学研究院营养科学研究所，上海 200031

摘要

本研究旨在探讨ErbB3结合蛋白1 (ErbB3-binding protein 1, Ebp1)在食管癌细胞生长中的作用及其机制。用携带Ebp1基因的慢病毒载体感染食管癌Eca109和KYSE150细胞，用real-time PCR检测食管癌组织中Ebp1 mRNA的表达情况，用MTT和结晶紫分别检测食管癌细胞生长和存活能力，用软琼脂细胞生长实验检测细胞克隆形成能力，用流式细胞术检测细胞凋亡率，用Western blot检测和凋亡有关的蛋白表达变化，用裸鼠皮下成瘤实验检测食管癌细胞的成瘤能力。结果显示，与配对的正常组织相比，食管癌组织中Ebp1 mRNA水平显著下降。过表达Ebp1不仅抑制食管癌细胞Eca109和KYSE150的体外生长和存活能力，而且能诱导这两种食管癌细胞发生凋亡，上调Rb和P53的蛋白表达，下调CyclinD1的表达。Ebp1过表达还能抑制Eca109细胞的裸鼠皮下成瘤能力。以上结果提示，Ebp1通过诱导细胞凋亡抑制食管癌细胞体外生长能力和体内成瘤能力。

关键词： ErbB3 结合蛋白1; 食管癌; 细胞生长; 凋亡; 成瘤

分类号：Q291

[Up-regulation of ErbB3-binding protein 1 inhibits the growth of esophageal carcinoma cells.] [Article in Chinese]

JIANG Hao, LIU Dong-Ping, XIE Dong, WEI Dong-Zhi

State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China； Institute of Nutrition Science, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China

Abstract

The objective of this study was to investigate the role of ErbB3-binding protein 1 (Ebp1) in the growth of esophageal squamous cell carcinoma (ESCC) cells and the underlying mechanism. Eca109 and KYSE150 cells were transfected with lentiviral vector carrying Ebp1 gene. The mRNA levels of Ebp1 in esophageal cancer tissues and paired adjacent normal tissues were examined by real-time PCR. The growth and viability of esophageal carcinoma cells were assessed using MTT and crystal violet assays, respectively. Clone-forming abilities of Eca109 and KYSE150 cells were analyzed by soft agar assay. Apoptotic rates of esophageal carcinoma cells were detected by flow cytometry, and expression levels of the proteins involved in apoptosis were assessed by Western blot. Tumorigenicity of Eca109 cells were detected by nude mouse transplantation tumor experiment. The results indicated that the mRNA levels of Ebp1 in esophageal cancer tissues was down-regulated compared with paired adjacent normal tissues. The growth and viability of Eca109 and KYSE150 cells were all suppressed by Ebp1 overexpression. Besides, Ebp1 overexpression induced apoptosis, increased Rb and P53 expressions, and decreased CyclinD1 expression in Eca109 and KYSE150 cells. In addition, Ebp1 overexpression inhibited the tumorigenesis of Eca109 cells in vivo. These results suggest that Ebp1 may suppress the growth of esophageal carcinoma cells in vitro and in vivo by inducing apoptosis.

Key words: ErbB3-binding protein 1 (Ebp1); esophageal carcinoma; cell growth; apoptosis; tumorigenesis

收稿日期：2016-08-22　　录用日期：2016-10-13

通讯作者：魏东芝　　E-mail: dzhwei@ecust.edu.cn

引用本文：

姜浩, 刘冬平, 谢东, 魏东芝. ErbB3结合蛋白1的上调抑制食管癌细胞生长[J]. 生理学报 2016; 68 (6): 740-746.

JIANG Hao, LIU Dong-Ping, XIE Dong, WEI Dong-Zhi. [Up-regulation of ErbB3-binding protein 1 inhibits the growth of esophageal carcinoma cells.] [Article in Chinese]. Acta Physiol Sin 2016; 68 (6): 740-746 (in Chinese with English abstract).