Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, School of Medicine, Northwest University, Xi'an, 710069, Shanxi, China.
College of Life Science, Northwest University, Xi'an, 710069, Shaanxi, China.
Mol Genet Genomics. 2023 Nov;298(6):1353-1364. doi: 10.1007/s00438-023-02046-w. Epub 2023 Jul 31.
The study was designed to detect the expression and clinical significance of the HEATR3 gene in bladder cancer (BCa) and to preliminarily explore whether this gene can affect the occurrence and development of BCa through the AKT/ERK signaling pathway. The expression and prognostic value of HEATR3 were explored based on The Cancer Genome Atlas (TCGA) and Genotypic Tissue Expression (GTEx) databases. Microarray immunohistochemical analysis was performed in 30 BCa cases to investigate the level of HEATR3 protein and to explore the relationship between HEATR3 and the clinicopathological features of BCa. Western Blot and qRT-PCR were used to detect HEATR3 protein and mRNA in BCa cell lines (5637, TCCSUP, SW780) and fallopian tube epithelial cell (SV-HUC-1). CCK8 method was employed to study the proliferation of BCa cells after heat treatment. Transwell assay was conducted to analyze the effect of HEATR3 on cell migration and invasion. And cell cycle and apoptosis were detected by flow cytometry. Furthermore, Western Blot assay was used to probe the effects of down-regulation of HEATR3 expression on the expression and phosphorylation levels of AKT and ERK proteins in BCa cells. Bioinformatics analysis showed that HEATR3 was significantly up-regulated in BCa, and high HEATR3 expression was associated with poor prognosis of BCa patients. In vitro experiments demonstrated that HEATR3 expression was up-regulated in BCa tissues compared with that in adjacent tissues. HEATR3 protein was also up-regulated in malignant cell lines. HEATR3 knockdown in BCa cells could inhibit cell proliferation, invasion and migration, block cell cycle and promote cell apoptosis. At the same time, HEATR3 knockdowns reduced the expression levels of p-AKT and p-ERK proteins. HEATR3 knockdown inhibits the development of BCa cells through the AKT/ERK signaling pathway. and it may become one of the most promising molecular targets for BCa treatment.
这项研究旨在检测 HEATR3 基因在膀胱癌（BCa）中的表达及其临床意义，并初步探讨该基因是否可以通过 AKT/ERK 信号通路影响 BCa 的发生和发展。基于癌症基因组图谱（TCGA）和基因型组织表达（GTEx）数据库探讨了 HEATR3 的表达和预后价值。对 30 例 BCa 病例进行了微阵列免疫组织化学分析，以研究 HEATR3 蛋白的水平，并探讨 HEATR3 与 BCa 临床病理特征之间的关系。Western Blot 和 qRT-PCR 用于检测 BCa 细胞系（5637、TCCSUP、SW780）和输卵管上皮细胞（SV-HUC-1）中 HEATR3 蛋白和 mRNA 的表达。CCK8 法研究了热处理后 BCa 细胞的增殖情况。Transwell 分析用于分析 HEATR3 对细胞迁移和侵袭的影响。并通过流式细胞术检测细胞周期和凋亡。此外，Western Blot 分析用于研究下调 HEATR3 表达对 BCa 细胞中 AKT 和 ERK 蛋白表达和磷酸化水平的影响。生物信息学分析表明，HEATR3 在 BCa 中显著上调，高 HEATR3 表达与 BCa 患者预后不良相关。体外实验表明，与相邻组织相比，HEATR3 在 BCa 组织中表达上调。HEATR3 蛋白在恶性细胞系中也上调。HEATR3 在 BCa 细胞中的敲低可抑制细胞增殖、侵袭和迁移，阻断细胞周期并促进细胞凋亡。同时，HEATR3 敲低降低了 p-AKT 和 p-ERK 蛋白的表达水平。HEATR3 敲低通过 AKT/ERK 信号通路抑制 BCa 细胞的发展，它可能成为 BCa 治疗最有前途的分子靶点之一。
