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DEP 结构域蛋白 1B 通过调节核孔蛋白 37 参与结肠癌细胞的增殖、转移、细胞周期停滞和凋亡。

DEPDC1B is involved in the proliferation, metastasis, cell cycle arrest and apoptosis of colon cancer cells by regulating NUP37.

机构信息

Department of General Surgery, The First Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330006, P.R. China.

Department of General Surgery, The First Affiliated Hospital of Nanchang University Jing'an County People's Hospital, Jing'an, Shanghai 330600, P.R. China.

出版信息

Mol Med Rep. 2023 Jun;27(6). doi: 10.3892/mmr.2023.13013. Epub 2023 May 19.

Abstract

It has been reported that DEP domain protein 1B (DEPDC1B) serves several roles in the occurrence and development of various types of cancer. Nevertheless, the effect of DEPDC1B on colorectal cancer (CRC), as well as its particular underlying molecular mechanism remain to be elucidated. In the present study, the mRNA and protein expression levels of DEPDC1B and nucleoporin 37 (NUP37) in CRC cell lines were assessed by reverse transcription‑quantitative PCR and western blotting, respectively. Cell Counting Kit‑8 and 5‑Ethynyl‑2'‑deoxyuridine assays were carried out to determine cell proliferation. In addition, the migration and invasion abilities of cells were evaluated using wound healing and Transwell assays. The changes in cell apoptosis and cell cycle distribution were assessed by flow cytometry and western blotting. Bioinformatics analysis and co‑immunoprecipitation assays were performed to predict and verify, respectively, the binding capacity of DEPDC1B on NUP37. The expression levels of Ki‑67 were detected by immunohistochemical assay. Finally, the activation of phosphoinositide 3‑kinase (PI3K)/protein kinase B (AKT) signaling was measured using western blotting. The results showed that DEPDC1B and NUP37 were upregulated in CRC cell lines. DEPDC1B and NUP37 silencing both inhibited the proliferation, migration and invasion capabilities of CRC cells and promoted cell apoptosis and cell cycle arrest. Furthermore, NUP37 overexpression reversed the inhibitory effects of DEPDC1B silencing on the behavior of CRC cells. Animal experiments demonstrated that DEPDC1B knockdown inhibited the growth of CRC in vivo by targeting NUP37. In addition, DEPDC1B knockdown inhibited the expression levels of the PI3K/AKT signaling‑related proteins in CRC cells and tissues by also binding to NUP37. Overall, the current study suggested that DEPDC1B silencing could alleviate the progression of CRC via targeting NUP37.

摘要

据报道,DEP 结构域蛋白 1B(DEPDC1B)在多种类型癌症的发生和发展中发挥多种作用。然而,DEPDC1B 对结直肠癌(CRC)的影响及其特定的潜在分子机制仍有待阐明。在本研究中,通过逆转录-定量 PCR 和 Western blot 分别评估了 CRC 细胞系中 DEPDC1B 和核孔蛋白 37(NUP37)的 mRNA 和蛋白表达水平。使用细胞计数试剂盒-8 和 5-乙炔基-2'-脱氧尿苷测定法来确定细胞增殖。此外,通过划痕愈合和 Transwell 测定评估了细胞的迁移和侵袭能力。通过流式细胞术和 Western blot 评估了细胞凋亡和细胞周期分布的变化。通过生物信息学分析和免疫共沉淀测定分别预测和验证了 DEPDC1B 与 NUP37 的结合能力。通过免疫组织化学检测 Ki-67 的表达水平。最后,使用 Western blot 测量了磷酸肌醇 3-激酶(PI3K)/蛋白激酶 B(AKT)信号的激活。结果显示,CRC 细胞系中 DEPDC1B 和 NUP37 表达上调。DEPDC1B 和 NUP37 沉默均抑制 CRC 细胞的增殖、迁移和侵袭能力,并促进细胞凋亡和细胞周期停滞。此外,NUP37 的过表达逆转了 DEPDC1B 沉默对 CRC 细胞行为的抑制作用。动物实验表明,通过靶向 NUP37,DEPDC1B 敲低抑制了 CRC 在体内的生长。此外,DEPDC1B 敲低还通过与 NUP37 结合抑制了 CRC 细胞和组织中 PI3K/AKT 信号相关蛋白的表达水平。总体而言,本研究表明,通过靶向 NUP37,DEPDC1B 沉默可减轻 CRC 的进展。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c871/10206622/ae46a20da05f/mmr-27-06-13013-g00.jpg

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