Abstract

This paper aims to discover the effect of Zinc Finger Protein 64 (ZFP64) and Notch pathway on lung adenocarcinoma cell. ZFP64 expression in cancer tissue and overall survival analysis was identified by TCGA-LUAD. ZFP64 expressions in tumor tissue (n = 30) and adjacent tissue (n = 30), and in human nontumorigenic bronchial epithelial cell line BEAS-2B and human lung adenocarcinoma cell lines (H23, H1975, H2228, and H2085) were measured via quantitative real-time polymerase chain reaction (qRT-PCR). H1975 cell viability, cell cycle progression, and migration after transfection or under Notch inhibitor MK-0752 treatment were detected through MTT assay, flow cytometer, and wound healing assay, respectively. Expressions of notch intracellular domain (NICD) and hairy and enhancer of split 1 (Hes-1) in H1975 cell were determined by western blot. Epithelial-mesenchymal transition (EMT)-related proteins (E-Cadherin and Vimentin) expressions were identified through qRT-PCR and western blot. ZFP64 expression in lung adenocarcinoma tissue and lung adenocarcinoma cell lines was higher and related to poor prognosis. After transfection, H1975 cell viability, migration, and expressions of Vimentin, NICD and Hes-1 were upregulated yet cell percentage in G0/G1 phase, E-cadherin expression was downregulated by overexpressed ZFP64. However, Notch inhibitor MK-0752 inhibited the effects of overexpressed ZFP64 on H1975 cell viability, cell cycle, migration, EMT progress, and Notch pathway activation. Overexpressed ZFP64 promoted the development of lung adenocarcinoma cells by activating Notch pathway.

摘要

本文旨在探索锌指蛋白64（ZFP64）和Notch通路对肺腺癌细胞的作用。TCGA-LUAD 鉴定了癌组织中 ZFP64 的表达和总生存分析。ZFP64 在肿瘤组织 ( n  = 30) 和邻近组织 ( n  = 30) 以及人非致瘤性支气管上皮细胞系 BEAS-2B 和人肺腺癌细胞系 (H23、H1975、H2228 和 H2085) 中的表达通过以下方式测量定量实时聚合酶链反应（qRT-PCR）。分别通过MTT法、流式细胞仪和伤口愈合法检测转染后或Notch抑制剂MK-0752处理后的H1975细胞活力、细胞周期进展和迁移。通过蛋白质印迹测定H1975细胞中notch细胞内结构域（NICD）和分裂1（Hes-1）的毛状和增强子的表达。通过 qRT-PCR 和蛋白质印迹鉴定上皮间质转化 (EMT) 相关蛋白 (E-钙粘蛋白和波形蛋白) 的表达。ZFP64在肺腺癌组织和肺腺癌细胞系中的表达较高，与预后不良有关。转染后H1975细胞活力、迁移和Vimentin、NICD和Hes-1表达上调，但在G0/G1期细胞百分比，E-cadherin 表达被过表达的 ZFP64 下调。然而，Notch 抑制剂 MK-0752 抑制过表达的 ZFP64 对 H1975 细胞活力、细胞周期、迁移、EMT 进展和 Notch 通路激活的影响。过表达的 ZFP64 通过激活 Notch 通路促进肺腺癌细胞的发展。