Department of Urology, The Second Affiliated Hospital of Nantong University, Nantong Clinical Research Center for Urological, Nantong, China.
Department of General, The Affiliated Hospital of Nantong University, Nantong, China.
Histol Histopathol. 2024 Oct;39(10):1343-1358. doi: 10.14670/HH-18-720. Epub 2024 Feb 9.
Dead-End 1 (DND1) is an RNA-binding protein (RBP) with regulatory functions in multiple cancers, including gastric and colorectal. Nevertheless, the role that DND1 plays in prostatic cancer (PCa) as well as the hidden molecular mechanism is still obscure. The gene expression of DND1 and survival analyses in PCa were analyzed by the UALCAN database. Expression of DND1 and chloride intracellular channel 4 (CLIC4) were detected by qRT-PCR and western blot analysis. The Cell Counting Kit-8 assay and EDU staining were employed for the estimation of cell viability. The capabilities of cells to migrate and invade were appraised by the wound healing assay as well as the Transwell assay, while epithelial-mesenchymal transition (EMT) was measured by immunofluorescence and western blot assay. The interaction of DND1 and CLIC4 was predicted by PCTA, linkedomics, and RPISeq databases. It was discovered that DND1 expression was elevated in PCa cells. DND1 silencing had suppressive impacts on the proliferative, migrative, and invasive capabilities as well as EMT in DU145 and 22Rv1 cells. Mechanistically, bioinformatic analysis demonstrated that DND1 was negatively correlated with CLIC4 and that DND1 protein could bind to CLIC4 mRNA. Additionally, the CLIC4 level was reduced in PCa cells. CLIC4 depletion countervailed the suppressive impacts of DND1 deficiency on the capabilities of DU145 and 22Rv1 cells to proliferate, migrate, and invade as well as the process of EMT. These results suggested that DND1 silencing repressed the proliferation, migration, invasion, and EMT in PCa by regulating the mRNA level of CLIC4.
DND1（无终止蛋白 1）是一种具有调节功能的 RNA 结合蛋白，在多种癌症中发挥作用，包括胃癌和结直肠癌。然而，DND1 在前列腺癌（PCa）中的作用以及隐藏的分子机制仍不清楚。通过 UALCAN 数据库分析了 DND1 的基因表达和 PCa 的生存分析。通过 qRT-PCR 和 Western blot 分析检测 DND1 和氯离子细胞内通道 4（CLIC4）的表达。通过细胞计数试剂盒-8 检测和 EDU 染色评估细胞活力。通过划痕愈合试验和 Transwell 试验评估细胞迁移和侵袭能力，通过免疫荧光和 Western blot 试验测量上皮-间充质转化（EMT）。通过 PCTA、linkedomics 和 RPISeq 数据库预测 DND1 和 CLIC4 的相互作用。结果发现，PCa 细胞中 DND1 表达上调。DND1 沉默对 DU145 和 22Rv1 细胞的增殖、迁移和侵袭能力以及 EMT 具有抑制作用。从机制上讲，生物信息学分析表明 DND1 与 CLIC4 呈负相关，DND1 蛋白可以与 CLIC4 mRNA 结合。此外，PCa 细胞中 CLIC4 水平降低。CLIC4 耗竭逆转了 DND1 缺乏对 DU145 和 22Rv1 细胞增殖、迁移、侵袭和 EMT 过程的抑制作用。这些结果表明，DND1 沉默通过调节 CLIC4 的 mRNA 水平抑制了 PCa 的增殖、迁移、侵袭和 EMT。
