Cui Mu, Li Zhen
School of Nursing, Xi'an Medical University, Xi'an, Shaanxi 710021, P.R. China.
Exp Ther Med. 2018 Jan;15(1):1048-1052. doi: 10.3892/etm.2017.5492. Epub 2017 Nov 10.
A previous study reported that Yes-associated protein (YAP) gene was overexpressed in esophageal squamous cell carcinoma (ESCC); however, the exact role of YAP in ESCC remains largely unclear. The present study aimed to investigate the effects of YAP inhibition on ESCC. In order to investigate the exact role of YAP in ESCC cells, a stable YAP low-expression ESCC cell line was established using YAP-small interfering RNA. MTT assay was performed to examine the cell proliferation ability, while flow cytometry were used to detect the cell apoptosis and cell cycle distribution. In addition, reverse transcription-quantitative polymerase chain reaction and western blot analysis were applied for mRNA and protein level detection, respectively. The results suggested that YAP gene inhibition significantly repressed the ECA-109 cell proliferation and induced cell apoptosis, whereas this inhibition had no significant effects on cell cycle. Furthermore, the expression levels of cell apoptosis-associated proteins were determined in the current study, and the data demonstrated that the B-cell lymphoma 2 (Bcl-2)/Bcl-2-associated X protein ratio and phosphorylated extracellular signal-regulated kinase expression were significantly reduced, while the p53 and caspase 3 levels were notably increased in YAP gene-inhibited ECA-109 cells. In conclusion, the current study revealed that YAP gene inhibition suppresses the proliferation and induces apoptosis in ECA-109 cells, indicating that the YAP gene serves as an oncogene in ESCC.
先前的一项研究报道,Yes相关蛋白(YAP)基因在食管鳞状细胞癌(ESCC)中过表达;然而,YAP在ESCC中的确切作用仍不清楚。本研究旨在探讨YAP抑制对ESCC的影响。为了研究YAP在ESCC细胞中的确切作用,使用YAP小干扰RNA建立了稳定的YAP低表达ESCC细胞系。采用MTT法检测细胞增殖能力,同时用流式细胞术检测细胞凋亡和细胞周期分布。此外,分别应用逆转录定量聚合酶链反应和蛋白质印迹分析进行mRNA和蛋白质水平检测。结果表明,YAP基因抑制显著抑制了ECA-109细胞的增殖并诱导细胞凋亡,而这种抑制对细胞周期没有显著影响。此外,本研究还测定了细胞凋亡相关蛋白的表达水平,数据表明,在YAP基因抑制的ECA-109细胞中,B细胞淋巴瘤2(Bcl-2)/Bcl-2相关X蛋白的比例以及磷酸化细胞外信号调节激酶的表达显著降低,而p53和半胱天冬酶3水平显著升高。总之,本研究表明YAP基因抑制可抑制ECA-109细胞的增殖并诱导其凋亡,这表明YAP基因在ESCC中起癌基因的作用。