Wang Haojie, Wang Yi, Liang Bing, He Fei, Li Yong, Che Jianbo, Li Xiaohui, Zhao Hui, Shi Gongning
Department of Cardiothoracic Surgery, Henan University Huaihe Hospital, Kaifeng, Henan 475000, China.
J Cancer Res Ther. 2016 Oct;12(Supplement):60-63. doi: 10.4103/0973-1482.191633.
The objective of this study was to investigate the expression pattern, functions, and possible mechanisms of RhoE/Rnd3, a novel member of the Rho GTPases family, in human esophageal squamous cell carcinoma (ESCC) cells by using molecular and cell-based experiments.
Quantitative polymerase chain reaction and Western blotting were carried out to determine the mRNA and protein expression of RhoE in ESCC cell lines, respectively. Both 3-(4,5-dimethylthiazol-2-yl)-2,5diphenyltetrazoliumbromide (MTT) and flow cytometry were applied to evaluate the effects of RhoE overexpression on ESCC cell growth and apoptosis. Furthermore, Western blotting was used to test the expression of epidermal growth factor receptor (EGFR) and phosphorylated-extracellular signal-regulated kinase (p-ERK) in ESCC cells after RhoE was forced and expressed.
RhoE was downregulated in human ESCC tissues. Overexpression of RhoE inhibited cell growth as assessed by MTT assay and induced apoptosis. Importantly, we proved that RhoE could negatively regulate the protein expression of EGFR and p-ERK, suggesting that RhoE might inhibit ESCC progression through the EGFR/ERK pathway.
Our data supported that RhoE could inhibit cell proliferation and promote apoptosis. Moreover, these tumor-suppressing effects might be acted through the negative regulation of EGFR/ERK signaling.
本研究旨在通过分子和细胞实验,探究Rho GTPases家族新成员RhoE/Rnd3在人食管鳞状细胞癌(ESCC)细胞中的表达模式、功能及可能机制。
分别采用定量聚合酶链反应和蛋白质免疫印迹法检测RhoE在ESCC细胞系中的mRNA和蛋白表达。运用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐(MTT)法和流式细胞术评估RhoE过表达对ESCC细胞生长和凋亡的影响。此外,在RhoE强制表达后,采用蛋白质免疫印迹法检测ESCC细胞中表皮生长因子受体(EGFR)和磷酸化细胞外信号调节激酶(p-ERK)的表达。
RhoE在人ESCC组织中表达下调。MTT法检测显示,RhoE过表达抑制细胞生长并诱导凋亡。重要的是,我们证实RhoE可负向调节EGFR和p-ERK的蛋白表达,提示RhoE可能通过EGFR/ERK途径抑制ESCC进展。
我们的数据支持RhoE可抑制细胞增殖并促进凋亡。此外,这些抑癌作用可能通过对EGFR/ERK信号的负调节发挥。