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微阵列分析显示 Tmub1 是大鼠肝细胞中与细胞周期相关的蛋白质。

Microarray analysis reveals Tmub1 as a cell cycle-associated protein in rat hepatocytes.

机构信息

Department of Hepatobiliary Surgery, Institute of Surgery Research, Daping Hospital, Third Military Medical University, Chongqing 400042, P.R. China.

Department of Hepatobiliary Surgery, 187 Military Hospital, Haikou, Hainan 571159, P.R. China.

出版信息

Mol Med Rep. 2018 Mar;17(3):4337-4344. doi: 10.3892/mmr.2018.8451. Epub 2018 Jan 17.

Abstract

Transmembrane and ubiquitin-like domain containing protein 1 (Tmub1), formerly known as hepatocyte odd protein shuttling (HOPS) has been recognized as a ubiquitously expressed shuttling protein that moves between the nucleus and cytoplasm in hepatocytes. Tmub1 is involved in liver regeneration and functions as a bridging protein in tumor cell proliferation. To investigate the transcriptional profile and potential biological processes affected by Tmub1 expression in normal rat hepatocytes, microarray and bioinformatics experiments were used to identify 127 mRNAs differentially expressed between Tmub1‑overexpression, Tmub1‑knockdown and normal BRL‑3A cells (fold‑change ≥2.5). The expression levels of 17 key node genes associated with the cell cycle were confirmed by reverse transcription-quantitative polymerase chain reaction analysis. Flow cytometry, 5‑Ethynyl‑20‑deoxyuridine, Cell Counting Kit‑8 and western blotting experiments revealed the effects on the cell cycle and the inhibition of proliferation in BRL‑3A cells overexpressing Tmub1. Further co‑immunoprecipitation assays demonstrated that Tmub1 interacts with cyclin A2 during the cell cycle and that the overexpression of Tmub1 may postpone cyclin A2 and cyclin B1 degradation in the M phase. The results of the present study indicated that Tmub1 functions as a cell proliferation inhibitor and cell cycle‑associated protein.

摘要

跨膜和泛素样结构域蛋白 1(TMUB1),以前称为肝细胞奇数蛋白穿梭(HOPS),已被认为是一种在核和细胞质之间穿梭的广泛表达的穿梭蛋白。TMUB1 参与肝脏再生,并作为肿瘤细胞增殖中的桥接蛋白发挥作用。为了研究 TMUB1 在正常大鼠肝细胞中的表达对转录谱和潜在生物学过程的影响,使用微阵列和生物信息学实验鉴定了 TMUB1 过表达、TMUB1 敲低和正常 BRL-3A 细胞之间差异表达的 127 个 mRNA(倍数变化≥2.5)。通过逆转录-定量聚合酶链反应分析证实了与细胞周期相关的 17 个关键节点基因的表达水平。流式细胞术、5-乙炔基-20-脱氧尿苷、细胞计数试剂盒-8 和蛋白质印迹实验揭示了 TMUB1 过表达对 BRL-3A 细胞细胞周期和增殖抑制的影响。进一步的共免疫沉淀实验表明,TMUB1 在细胞周期中与细胞周期蛋白 A2 相互作用,并且 TMUB1 的过表达可能会延迟 M 期细胞周期蛋白 A2 和细胞周期蛋白 B1 的降解。本研究的结果表明,TMUB1 作为细胞增殖抑制剂和细胞周期相关蛋白发挥作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ed76/5802207/b44457384b57/MMR-17-03-4337-g00.jpg

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