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本研究旨在探讨Erb B3结合蛋白1(Erb B3-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 m RNA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 m RNA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Cyclin D1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。
This study aimed to investigate the role of Erb B3-binding protein 1 (Ebp1) in esophageal cancer cell growth and its mechanism. Eca109 and KYSE150 cells were infected with lentiviral vector carrying Ebp1 gene. The expression of Ebp1 m RNA in esophageal cancer tissues was detected by real-time PCR. The growth and viability of esophageal cancer cells were detected by MTT and crystal violet. The ability of colony formation was detected by agar cell growth assay. The apoptosis rate was detected by flow cytometry. The protein expression of apoptosis was detected by Western blot. The tumorigenic ability of esophageal cancer cells was detected by subcutaneous tumor formation assay in nude mice. The results showed that the level of Ebp1 m RNA in esophageal cancer tissues was significantly decreased compared with the matched normal tissues. Overexpression of Ebp1 not only inhibited the growth and survival of esophageal cancer cells Eca109 and KYSE150 in vitro, but also induced the apoptosis of these two esophageal cancer cells, up-regulated the protein expressions of Rb and P53 and down-regulated the expression of Cyclin D1. Ebp1 overexpression also inhibited the subcutaneous tumorigenicity of Eca109 cells in nude mice. The above results suggest that Ebp1 inhibits the growth of esophageal cancer cells in vitro and the ability of tumorigenesis in vivo by inducing apoptosis.