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E2FBP1 antagonizes the p16^(INK4A)-Rb tumor suppressor machinery for growth suppression and cellular senescence by regulating promyelocytic leukemia protein stability

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International Journal of Oral Science 2011年第4期3卷 200-208页

作者： Yayoi Fukuyo Akiko Takahashi Eiji Hara Nobuo Horikoshi Tej K. Pandita Takuma Nakajima Department of Immunology Tulane National Primate Research Center Section of Bacterial Pathogenesis Graduate School Tokyo Medical and Dental University Division of Cancer Biology The Cancer Institute Japanese Foundation for Cancer Research Division of Molecular Radiation Biology Department of Radiation Oncology University of Texas Southwestern Medical Center

Cellular senescence is an irreversible cell cycle arrest triggered by the activation of oncogenes or mitogenic signaling as well as the enforced expression of tumor suppressors such as p53, p16INK4A and promyelocytic leukemia protein （PML） in normal cells. E2F-binding protein 1 （E2FBP1）, a transcription regulator for E2F, induces PML reduction and suppresses the formation of PML-nuclear bodies, whereas the down-regulation of E2FBP1 provokes the PML-dependent premature senescence in human normal fibroblasts. Here we report that the depletion of E2FBP1 induces the accumulation of PML through the Ras-dependent activation of MAP kinase signaling. The cellular levels of p16INK4A and p53 are elevated during premature senescence induced by depletion of E2FBP1, and the depletion of p16INK4A, but not p53 rescued senescent cells from growth arrest. Therefore, the premature senescence induced by E2FBP1 depletion is achieved through the pl6INK4A-Rb pathway. Similar to human normal fibroblasts, the growth inhibition induced by E2FBP1 depletion is also observed in human tumor cells with intact p16INK4A and Rb. These results suggest that E2FBP1 functions as a critical antagonist to the pI6INK4A-Rb tumor suppressor machinery by regulating PML stability.

关键词： E2F-binding protein 1 senescence cell cycle ubiquitin promyelocytic leukemia protein

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Transfection of promyelocytic leukemia in retrovirus vector inhibits growth of human bladder cancer cells

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Acta Pharmacologica Sinica 2005年第5期26卷 610-615页

作者： Lei LI,Da-lin HE~2 Institute of Urology,First Hospital of Xi'an Jiaotong University,Xi'an 710061,China Key Laboratory of Environment and Genes Related to Diseases of Ministry of Education,Xi'an Jiaotong University,Xi'an 710061,China Institute of Urology First Hospital of Xi'an Jiaotong University Xi'an China Key Laboratory of Environment and Genes Related to Diseases of Ministry of Education Xi'an Jiaotong University Xi'an China

Aim:To construct a recombinant retrovirus vector carrying human promyelocyticleukemia(PML)cDNA and identify its expression and biology role in bladdercancer UM-UC-2 cells for future gene ***:PML full-length cDNAwas inserted into the EcoR I and BamH I site of pLXSN vector containing the longterminal repeat(LTR)*** vector was identified by restriction enzymedigestion and then transfected into PA317 packaging cell line by calcium phos-phate *** cDNA was detected by polymerase chain reaction(PCR)and the protein was identified by laser confocal microscopy and Westernblot in bladder cancer cells,*** morphology was observed byinverted phase contrast microscope,and MTT assay determined growth curve ofthe bladder cancer ***:Restriction enzyme digestion proved that a 2.1kb PML cDNA was inserted into the pLXSN *** assay demonstrated that304 bp fragments were found in UM-UC-2/pLPMLSN *** confocalmicroscopy showed speck dots fluorescence in the UM-UC2/pLPMLSN nucleus.A 90 kD specific brand was found by Western *** assay demonstrated theUM-UC-2/pLPMLSN bladder cancer growth ***:The retroviruspLPMLSN vector was successfully constructed and could generate high effec-tive expression of human PML in bladder cancer cell UM-UC-2,suggesting thatPML recombinant retrovirus have potential utility in the gene therapy for bladdercancer.

关键词： promyelocytic leukemia protein retrovirus gene therapy

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