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Ectopic expression of (Mm)Kin17 protein inhibits cell proliferation of human tumor-derived cells
D S Biard1, P Kannouche, C Lannuzel-Drogou
1Laboratoire de Génétique de la Radiosensibilité, DSV-DRR, Fontenay aux Roses, 92265, France. biard@dsvidf.cea.fr
Abstract:
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.
Insights
Kin17 protein overexpression significantly reduced cancer cell proliferation and altered cell growth and morphology in established cells. This suggests Kin17 may interfere with DNA replication processes.
Area of Science:
- Molecular Biology
- Cell Biology
- Cancer Research
Background:
- Kin17 protein is a mammalian nuclear protein involved in DNA damage response and binds to curved DNA.
- Its precise biological role, particularly in cell proliferation and morphology, requires further elucidation.
Purpose of the Study:
- To investigate the biological role of Kin17 protein through overexpression studies in various cell lines.
- To determine the impact of Kin17 on cell proliferation, morphology, and potential interference with DNA replication.
Main Methods:
- Construction and transfection of EBV-derived vectors carrying Mus musculus (Mm)Kin17 cDNA into tumorigenic (HeLa, H1299, HCT116) and immortalized (HEK 293) cell lines.
- Analysis of cell proliferation, plating efficiency, cell morphology, and nuclear structures following (Mm)Kin17 expression.
- Isolation and characterization of stable (Mm)Kin17-expressing cell clones.
Main Results:
- Overexpression of (Mm)Kin17 drastically reduced proliferation in all tested tumorigenic cell lines, irrespective of their p53 status.
- In HEK 293 cells, (Mm)Kin17 expression led to transient cell populations and altered morphology, including multinucleation and multilobed nuclei.
- Stable (Mm)Kin17 expression in HEK 293 clones (B223.1, B223.2) resulted in decreased proliferative capacity, plating efficiency, and altered cell shape.
Conclusions:
- (Mm)Kin17 overexpression inhibits proliferation in tumorigenic cells and affects cell growth and morphology in immortalized cells.
- The observed effects suggest that Kin17 protein may play a role in regulating DNA replication.
- Kin17's impact on cell proliferation is independent of p53 status, highlighting its potential as a therapeutic target.