Department of Obstetrics and Gynecology, Nagoya University Graduate School of Medicine, 65, Tsurumai-cho, Showa-ku, Nagoya, Aichi, 466-8550, Japan.
Bell Research Center, Department of Obstetrics and Gynecology Collaborative Research, Nagoya University Graduate School of Medicine, Nagoya, Japan.
Reprod Sci. 2023 Sep;30(9):2829-2841. doi: 10.1007/s43032-023-01230-y. Epub 2023 Apr 4.
This study was aimed to identify a novel metastasis-promoting molecule and elucidate its functional and prognostic roles in cervical cancer. DDIT4 (DNA-damage-inducible transcript 4), a hypoxia-inducible gene, was identified by analyzing multiple microarray databases. The correlation between DDIT4 expression in immunohistochemistry and clinicopathological characteristics in the public database and our cohort was evaluated by statistical analysis. Transwell® assay and wound-healing assay to determine cell migration and invasion were performed. DDIT4 was knocked down using siRNA or lentiviral vectors. The potential downstream pathways of DDIT4 were explored and verified by a gene set enrichment analysis and western blotting. The in vivo metastatic capability was determined with the use of an intraperitoneal injection mouse model. In the analysis of the public database and our cohort, DDIT4 high expression was significantly related to short overall survival and lymph node metastasis in patients with early-stage cervical cancer. The knockdown of DDIT4 attenuated the migration and invasion activity of tumor cells in vitro and reduced the expression of epithelial-mesenchymal transition (EMT)-related proteins and the NF-κB pathway in cervical cancer cells. DDIT4 also promoted tumor progression in the mouse model. Our results indicate that DDIT4 can be a prognostic indicator in cervical cancer and promote lymph node metastasis, augmenting malignancy via the EMT and NF-kB pathways.
本研究旨在鉴定一种新型的促进转移的分子，并阐明其在宫颈癌中的功能和预后作用。通过分析多个微阵列数据库，鉴定出 DDIT4（DNA 损伤诱导转录物 4），这是一种缺氧诱导基因。通过统计学分析评估了公共数据库和我们队列中 DDIT4 在免疫组织化学中的表达与临床病理特征的相关性。通过 Transwell®测定和划痕愈合测定来确定细胞迁移和侵袭。使用 siRNA 或慢病毒载体敲低 DDIT4。通过基因集富集分析和 Western blot 来探索和验证 DDIT4 的潜在下游途径。使用腹腔注射小鼠模型来确定体内转移能力。在公共数据库和我们队列的分析中，DDIT4 高表达与早期宫颈癌患者的总生存期短和淋巴结转移显著相关。DDIT4 的敲低减弱了肿瘤细胞在体外的迁移和侵袭活性，并降低了宫颈癌细胞中上皮-间充质转化（EMT）相关蛋白和 NF-κB 通路的表达。DDIT4 还促进了小鼠模型中的肿瘤进展。我们的结果表明，DDIT4 可以作为宫颈癌的预后指标，并通过 EMT 和 NF-kB 通路促进淋巴结转移，增加恶性程度。
