Wild type p53 gene causes reorganization of cytoskeleton and, therefore, the impaired deformability and difficult migration of murine erythroleukemia cells.
We studied the role of p53 gene in the biophysics and biology in murine erythroleukemia cell line (MEL), with the goal of understanding the influence of this tumor suppressor gene on the deformability and metastasis of tumor cells. Experiments were performed on MEL and p53-transfected MEL (MEL-M with mutant p53 gene and MEL-W with wild-type p53 gene). The cell growth curves indicated that the over-expression of wild-type p53 gene significantly suppressed the growth of MEL, with G(0)-G(1) arrest and apoptosis shown by flow cytometric assays. Confocal laser scanning microscopy revealed that the MEL-W had a more compact organization of the F-Actin cytoskeleton than MEL and MEL-M. Fluorescence polarization measurement indicated a higher membrane fluidity of MEL-W than the other two groups. Fourier transform infrared spectroscopy (FT-IR) showed changes in the composition and/or structure of membrane lipids in MEL-W, with decreases in secondary structures of proteins such as alpha-helix, turns and bends and random coil, in comparison to MEL and MEL-M. The osmotic fragility curves indicated that MEL-W was more fragile and micropipette experiments showed that they had increased elasticity and reduced deformability in comparison to MEL and MEL-M. The adhesion assay with the use of the flow chamber revealed a lower adhesion rate of MEL-W to endothelial cells at high shear stress. The present study on the molecular biology with biophysics of MEL cells contributes to our knowledge on the tumor suppressor gene p53.
Yao
Weijuan
W
School of Basic Medical Sciences, Peking University, Beijing, Peoples Republic of China.
Gu
Li
L
Sun
Dagong
D
Ka
Weibo
W
Wen
Zongyao
Z
Chien
Shu
S
D000199:Actins / Q000032:analysis / Q000378:metabolism; D000818:Animals; D017209:Apoptosis / Q000502:physiology; D015153:Blotting, Western; D002448:Cell Adhesion / Q000502:physiology; D002453:Cell Cycle / Q000502:physiology; D002455:Cell Division / Q000502:physiology; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000502:physiology*; D003599:Cytoskeleton / Q000502:physiology*; D004548:Elasticity; D042783:Endothelial Cells / Q000502:physiology; D005434:Flow Cytometry; D005454:Fluorescence Polarization; D016158:Genes, p53 / Q000235:genetics / Q000502:physiology*; D006801:Humans; D004915:Leukemia, Erythroblastic, Acute / Q000473:pathology; D008560:Membrane Fluidity / Q000502:physiology; D051379:Mice; D018613:Microscopy, Confocal; D009154:Mutation; D009996:Osmotic Fragility / Q000502:physiology; D017550:Spectroscopy, Fourier Transform Infrared; D014162:Transfection
D000199:Actins