Lu Xiaowei, Hu Kui, Zhang Dandan, Yin Xuefeng, Nie Jifeng, Zhao Kai
Department of General Surgery, Zhejiang Integrated Traditional and Western Medicine Hospital, Hangzhou, 310009, Zhejiang, China.
Department of Emergency Surgery, Zhejiang Integrated Traditional and Western Medicine Hospital, Hangzhou, 310009, Zhejiang, China.
J Cancer Res Clin Oncol. 2024 Dec 20;151(1):11. doi: 10.1007/s00432-024-06059-9.
Krüppel-like factor 5 (KLF5) is recognized as a tumor mediator in multiple types of tumors. Nevertheless, whether KLF5 plays a role in gallbladder cancer (GBC) remains to be elucidated. This study aims to clarify the role of KLF5 in the proliferation, migration and angiogenesis in GBC cells.
The expressions of KLF5 and platelet-derived growth factor subunit A (PDGFA) in GBC cell lines were analyzed by qRT-PCR and western blot assay. Cell proliferation was assessed utilizing the Cell Counting Kit-8 assay and EDU staining. Cell apoptosis was evaluated using flow cytometry, and apoptosis-related proteins was examined by western blotting. The migratory and invasive capabilities were evaluated utilizing wound healing and Transwell. Angiogenesis was assessed by ELISA, tube formation assay and western blot. The interaction between KLF5 and PDGFA was confirmed by ChIP assay, as well as luciferase reporter assay.
In this study, we discovered that the levels of KLF5 and PDGFA were upregulated in GBC cells. Silencing of KLF5 reduced the viability and suppressed the proliferation of GBC cells, as well as promoting the apoptosis. In addition, KLF5 silencing restrained the invasion and migration and angiogenesis in NOZ and GBC-SD cells. KLF5 transcription activated PDGFA expression and KLF5 was proved to bind to PDGFA promoter in NOZ cells. Following the silencing of PDGFA, the proliferation, invasion, migration, angiogenesis and apoptosis exhibited similar changes to KLF5 silencing. Additionally, PDGFA overexpression reversed the effects of KLF5 silencing on NOZ cells.
Collectively, our results suggest that KLF5 regulates GBC cell proliferation, invasion, migration, angiogenesis, as well as apoptosis, via mediating PDGFA transcriptionally, which might provide a novel therapeutic strategy for treatment of human GBC.
Krüppel样因子5（KLF5）在多种肿瘤中被认为是一种肿瘤调节因子。然而，KLF5在胆囊癌（GBC）中是否发挥作用仍有待阐明。本研究旨在明确KLF5在GBC细胞增殖、迁移和血管生成中的作用。
通过qRT-PCR和蛋白质免疫印迹法分析KLF5和血小板衍生生长因子A亚基（PDGFA）在GBC细胞系中的表达。利用细胞计数试剂盒-8法和EDU染色评估细胞增殖。采用流式细胞术评估细胞凋亡，并通过蛋白质免疫印迹法检测凋亡相关蛋白。利用伤口愈合实验和Transwell实验评估细胞的迁移和侵袭能力。通过酶联免疫吸附测定法、管腔形成实验和蛋白质免疫印迹法评估血管生成。通过染色质免疫沉淀实验以及荧光素酶报告基因实验证实KLF5与PDGFA之间的相互作用。
在本研究中，我们发现GBC细胞中KLF5和PDGFA的水平上调。沉默KLF5可降低GBC细胞的活力，抑制其增殖，并促进细胞凋亡。此外，沉默KLF5可抑制NOZ和GBC-SD细胞的侵袭、迁移和血管生成。KLF5转录激活PDGFA的表达，并且在NOZ细胞中证实KLF5与PDGFA启动子结合。沉默PDGFA后，细胞的增殖、侵袭、迁移、血管生成和凋亡表现出与沉默KLF5时相似的变化。此外，PDGFA过表达可逆转沉默KLF5对NOZ细胞的影响。
总体而言，我们的结果表明KLF5通过转录调控PDGFA来调节GBC细胞的增殖、侵袭、迁移、血管生成以及凋亡，这可能为人类GBC的治疗提供一种新的治疗策略。
