Ectopic expression of (Mm)Kin17 protein inhibits cell proliferation of human tumor-derived cells.
To characterize the biological role of Kin17 protein, a mammalian nuclear protein which participates in the response to UV and ionizing radiation and binds to curved DNA, EBV-derived vectors carrying (Mm)Kin17 cDNA were constructed and transfected in tumorigenic cells harboring different p53 profiles (HeLa, H1299, and HCT116) and in immortalized HEK 293 cells. (Mm)Kin17 protein expression induced a tremendous decrease in cell proliferation of the three tumorigenic cell lines 2 weeks after transfection. Transfection of HEK 293 cells with an pEBVCMV(Mm)Kin17 plasmid gave rise to numerous (Mm)Kin17-expressing cells which constantly disappeared with time, preventing the establishment of (Mm)Kin17-expressing cells. Several independent clones were isolated from HEK 293 cells carrying a pEBVMT(Mm)Kin17 vector. The two clones described here (B223.1 and B223.2) exhibited different (Mm)Kin17 protein levels and displayed a gradual decrease in their proliferative capacities. In B223.1 cells, the basal expression of (Mm)Kin17 greatly reduced plating efficiency and cell growth. B223.1 cell morphology was altered, with numerous round-shaped cells whose spreading on the culture support was hampered. We observed giant multinucleated cells or cells containing micronuclei-like structures and/or multilobed nuclei. To conclude, (Mm)Kin17 overexpression reduced the proliferation of tumorigenic cells independently of their p53 status and modified cell growth and cell morphology of established HEK 293 cells producing (Mm)Kin17 protein. It is likely that (Mm)Kin17 may interfere with DNA replication.
Laboratoire de Génétique de la Radiosensibilité, DSV-DRR, Fontenay aux Roses, 92265, France. biard@dsvidf.cea.fr
D S
DS
Biard
P
P
Kannouche
C
C
Lannuzel-Drogou
P
P
Mauffrey
F
F
Apiou
J F
JF
Angulo
D000818:Animals; D015153:Blotting, Western; D002104:Cadmium / Q000494:pharmacology; D002452:Cell Count; D002455:Cell Division; D002461:Cell Line, Transformed; D048429:Cell Size; D002999:Clone Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D019926:Cyclin B / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics / Q000502:physiology*; D019084:Fluorescent Antibody Technique, Indirect; D015870:Gene Expression / Q000187:drug effects; D015726:Giant Cells / Q000378:metabolism; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D051379:Mice; D009687:Nuclear Proteins; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism; D016601:RNA-Binding Proteins; D014162:Transfection; D014407:Tumor Cells, Cultured; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism; D015032:Zinc / Q000494:pharmacology
D019926:Cyclin B; D004268:DNA-Binding Proteins; C489487:KIN protein, human; D009687:Nuclear Proteins; D018809:Proliferating Cell Nuclear Antigen; D016601:RNA-Binding Proteins; D016159:Tumor Suppressor Protein p53; D002104:Cadmium; C071214:Kin protein, mouse; D015032:Zinc