Microarray analysis reveals Tmub1 as a cell cycle-associated protein in rat hepatocytes
نویسندگان
چکیده
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.
منابع مشابه
IL-6 induction of hepatocyte proliferation through the Tmub1-regulated gene pathway.
The expression of transmembrane and ubiquitin-like domain containing 1 (Tmub1) is upregulated during liver regeneration, however, the function and underlying molecular mechanisms responsible for Tmub1 action remain to be determined. This study utilized BRL-3A rat liver cells for Tmub1 shRNA lentivirus infection and IL-6 stimulation. Semi-quantitative RT-PCR and western blot analysis were used t...
متن کاملIron Overload Induced Apoptotic Cell Death in Isolated Rat Hepatocytes Mediated by Reactive Oxygen Species
Isolated rat hepatocytes in culture were incubated with different concentrations of iron-sorbitol (50, 100, 150, and 200 µM) to assess the changes in reactive oxygen species (ROS) and lipid peroxidation leading to apoptotic hepatocyte cell death. The viability of hepatocytes was declined depending on the iron concentration. One hour incubation of the cells with 100 µM iron resulted in decreased...
متن کاملIron Overload Induced Apoptotic Cell Death in Isolated Rat Hepatocytes Mediated by Reactive Oxygen Species
Isolated rat hepatocytes in culture were incubated with different concentrations of iron-sorbitol (50, 100, 150, and 200 µM) to assess the changes in reactive oxygen species (ROS) and lipid peroxidation leading to apoptotic hepatocyte cell death. The viability of hepatocytes was declined depending on the iron concentration. One hour incubation of the cells with 100 µM iron resulted in decreased...
متن کاملEffects of Over-Expression of LOC92912 Gene on Cell Cycle Progression
Background: We had previously identified the genes involved in squamous cell carcinoma of the head and neck using differential display and DNA microarray techniques. We also reported the first analytical study on a novel human gene called LOC92912, which was identified by differential display as a gene up-regulated in such carcinomas. LOC92912, which is a putative member of the E2 ubiquitin con...
متن کاملTransmembrane and Ubiquitin-Like Domain Containing 1 (Tmub1) Regulates Locomotor Activity and Wakefulness in Mice and Interacts with CAMLG
Tmub1 (C7orf21/HOPS) encodes a protein containing a ubiquitin-like domain. Tmub1 is highly expressed in the nervous system. To study its physiological function, we generated mice with Tmub1 deleted by homologous recombination. The knockout mice were grossly normal and viable. In a comprehensive behavioral testing battery, the only knockout phenotype displayed was a strong increase in home cage ...
متن کامل