{"doi":"10.1002/mc.10111","title":"Inhibition of proteasome‐dependent degradation of Wee1 in G<sub>2</sub>‐arrested Hep3B cells by TGFβ1","abstract":"<jats:title>Abstract</jats:title><jats:p>Transforming growth factor β1 (TGFβ1)‐induced G<jats:sub>2</jats:sub> arrest was observed when a proliferation inhibitory function of the retinoblastoma protein (Rb) was compromised, but the mechanism underlying the G<jats:sub>2</jats:sub> arrest was poorly characterized compared with that of G<jats:sub>1</jats:sub> arrest. In the present study, we characterized G<jats:sub>2</jats:sub> arrest induced by TGFβ1 (1 ng/mL) in the Rb‐negative hepatoma cell line (Hep3B) and compared with G<jats:sub>1</jats:sub> arrest in the Rb‐positive hepatoma cell line (Huh7). Activities of cyclin‐dependent kinases (CDK) 2 and cell division cycle (CDC) 2 were markedly decreased at 24 h, the time when cell‐cycle arrest became apparent in both cell lines. However, considerable amounts of inactive CDC2‐cyclinB1 complexes were present in the nucleus of G<jats:sub>2</jats:sub>‐arrested Hep3B but were not present in G<jats:sub>1</jats:sub>‐arrested Huh7. The inhibitory phosphorylation of CDC2 on Tyr‐15 was significantly elevated at 12–24 h, and its levels gradually declined during G<jats:sub>2</jats:sub> arrest in Hep3B. In particular, augmentation of CDK inhibitors p21<jats:sup>cip1</jats:sup> and p27<jats:sup>kip1</jats:sup> and Wee1 kinase and diminution of CDC25C phosphatase coincided with induced Tyr‐15 phosphorylation and inhibition of CDC2. Wee1 in Hep3B was unstable and was degraded in a proteasome‐dependent manner, but it became substantially stabilized within 6 h of TGFβ1 treatment. Moreover, a Wee1 inhibitor, PD0166285, abrogated the TGFβ1‐induced G<jats:sub>2</jats:sub> arrest in Hep3B. These findings suggest that TGFβ1 induced G<jats:sub>2</jats:sub> arrest in Hep3B at least in part through stabilization of Wee1 and subsequent increase in Tyr‐15 phosphorylation and inhibition of CDC2. © 2003 Wiley‐Liss, Inc.</jats:p>","journal":"Molecular Carcinogenesis","year":2003,"id":632937,"datarank":0.5709993734655481,"base_score":3.8066624897703196,"endowment":3.8066624897703196,"self_citation_contribution":0.5709993734655481,"citation_network_contribution":0.0,"self_endowment_contribution":0.5709993734655481,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":44,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":null,"is_data_producer":false,"deposit_databanks":null,"is_oa":false,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":null,"fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":1583771,"name":"Takato Ueno","orcid":null,"position":1,"is_corresponding":false},{"id":1640816,"name":"Rina Kimura","orcid":null,"position":2,"is_corresponding":false},{"id":1566908,"name":"Motoaki Ohtsubo","orcid":null,"position":3,"is_corresponding":false},{"id":13849,"name":"Toru Nakamura","orcid":"0000-0001-5752-0814","position":4,"is_corresponding":false},{"id":1572789,"name":"Hironori Koga","orcid":null,"position":5,"is_corresponding":false},{"id":1583770,"name":"Takuji Torimura","orcid":null,"position":6,"is_corresponding":false},{"id":1640825,"name":"Sanae Uchida","orcid":null,"position":7,"is_corresponding":false},{"id":1640829,"name":"Katsumi Yamashita","orcid":null,"position":8,"is_corresponding":false},{"id":1583778,"name":"Michio Sata","orcid":null,"position":9,"is_corresponding":false},{"id":1640813,"name":"Osamu Hashimoto","orcid":null,"position":0,"is_corresponding":false}],"reference_count":0,"raw_metadata":{"has_enrichment":true,"resolved":true,"title":"Inhibition of proteasome‐dependent degradation of Wee1 in G<sub>2</sub>‐arrested Hep3B cells by TGFβ1","abstract":"<jats:title>Abstract</jats:title><jats:p>Transforming growth factor β1 (TGFβ1)‐induced G<jats:sub>2</jats:sub> arrest was observed when a proliferation inhibitory function of the retinoblastoma protein (Rb) was compromised, but the mechanism underlying the G<jats:sub>2</jats:sub> arrest was poorly characterized compared with that of G<jats:sub>1</jats:sub> arrest. In the present study, we characterized G<jats:sub>2</jats:sub> arrest induced by TGFβ1 (1 ng/mL) in the Rb‐negative hepatoma cell line (Hep3B) and compared with G<jats:sub>1</jats:sub> arrest in the Rb‐positive hepatoma cell line (Huh7). Activities of cyclin‐dependent kinases (CDK) 2 and cell division cycle (CDC) 2 were markedly decreased at 24 h, the time when cell‐cycle arrest became apparent in both cell lines. However, considerable amounts of inactive CDC2‐cyclinB1 complexes were present in the nucleus of G<jats:sub>2</jats:sub>‐arrested Hep3B but were not present in G<jats:sub>1</jats:sub>‐arrested Huh7. The inhibitory phosphorylation of CDC2 on Tyr‐15 was significantly elevated at 12–24 h, and its levels gradually declined during G<jats:sub>2</jats:sub> arrest in Hep3B. In particular, augmentation of CDK inhibitors p21<jats:sup>cip1</jats:sup> and p27<jats:sup>kip1</jats:sup> and Wee1 kinase and diminution of CDC25C phosphatase coincided with induced Tyr‐15 phosphorylation and inhibition of CDC2. Wee1 in Hep3B was unstable and was degraded in a proteasome‐dependent manner, but it became substantially stabilized within 6 h of TGFβ1 treatment. Moreover, a Wee1 inhibitor, PD0166285, abrogated the TGFβ1‐induced G<jats:sub>2</jats:sub> arrest in Hep3B. These findings suggest that TGFβ1 induced G<jats:sub>2</jats:sub> arrest in Hep3B at least in part through stabilization of Wee1 and subsequent increase in Tyr‐15 phosphorylation and inhibition of CDC2. © 2003 Wiley‐Liss, Inc.</jats:p>","is_dataset_classified":null,"base_score":0.0,"endowment":0.0,"datacite_reuse_total":0,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"12669309","pmcid":null,"openalex_id":null,"authors":[],"funders":[],"total_grants":0,"fwci":null,"citation_percentile":null,"influential_citations":0,"citation_trend":[],"oa_status":"closed","license":"http://onlinelibrary.wiley.com/termsAndConditions#vor","oa_locations":[{"url":"https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1002%2Fmc.10111","host_type":"publisher"},{"url":"https://onlinelibrary.wiley.com/doi/pdf/10.1002/mc.10111","host_type":"publisher"}],"fields_of_study":["Base Sequence","Carcinoma, Hepatocellular","Cell Cycle","Cell Cycle Proteins","Cell Division","Cysteine Endopeptidases","DNA Primers","G2 Phase","Humans","Kinetics","Liver Neoplasms","Multienzyme Complexes","Nuclear Proteins","Phosphorylation","Proteasome Endopeptidase Complex","Protein-Tyrosine Kinases","Reverse Transcriptase Polymerase Chain Reaction","Transforming Growth Factor beta","Transforming Growth Factor beta1","Tumor Cells, Cultured"],"mesh_terms":["Tumor Cells, Cultured","Humans","Carcinoma, Hepatocellular","Liver Neoplasms","Multienzyme Complexes","Proteasome Endopeptidase Complex","Cysteine Endopeptidases","Transforming Growth Factor beta","Cell Cycle Proteins","Nuclear Proteins","DNA Primers","Reverse Transcriptase Polymerase Chain Reaction","Cell Cycle","Cell Division","G2 Phase","Base Sequence","Phosphorylation","Kinetics","Transforming Growth Factor beta1","Protein-Tyrosine Kinases"],"keywords":[],"sdg_mappings":[],"linked_datasets":[],"clinical_trials":[],"software_tools":[],"database_accessions":[],"source":"live","citation_network_status":"fetched"},"created_at":"2026-08-06T10:37:22.384853Z","pmid":null,"pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}