The expressions of intrinsic and extrinsic apoptotic pathway proteins in neutrophils of oral cavity cancer patients: a preliminary study.
BACKGROUND
The biological availability and activity of polymorphonuclear neutrophils (PMNs) are regulated by their short life span, which can be additionally shortened by a malignant process. The signaling pathways leading to apoptotic PMN death are classified in two categories: the intrinsic and the extrinsic. In the present study the expressions of proteins participating in the extrinsic apoptotic pathway (DR5, FADD, caspase-8 activity) and the intensity of apoptosis of PMNs from patients with cancer of the oral cavity were examined. The expression of proteins participating in the intrinsic pathway (Bax and Mcl-1) were also examined in these cells. The results can be helpful in explaining the reasons for the decreased activity of these cells in oral cavity cancer patients.


METHODS
The examinations were carried out in patients with squamous cell carcinoma of the oral cavity before and after treatment. The expressions of all the proteins were measured in neutrophils and, for comparison, in autologous peripheral blood mononuclear cells (PBMCs). Western blot analysis was used to assay the expressions of DR5, FADD, Bax, and Mcl-1 in cell lysates. The apoptosis level was determined by flow cytometry and caspase-8 activity by colorimetric assay.


RESULTS
A lack of changes in DR5 expression associated with increased FADD protein expression and caspase-8 activity accompanied the accelerated apoptosis rates in the PMNs of the patients before treatment. Decreased expression of anti-apoptotic Mcl-1 protein was associated with an unchanged expression of pro-apoptotic Bax protein. There were no such changes in the patients PBMCs. Increased expression of Mcl-1 in the PMNs of the patients following surgical treatment was found.


CONCLUSIONS
The acceleration of the apoptosis of PMNs of oral cavity cancer patients before treatment is dependent on both the intrinsic and extrinsic pathways.
Jabłońska
Ewa
E
Department of Immunology, Medical University of Białystok, 15-274 Białystok, Poland. ewaj@amb.edu.pl
Garley
Marzena
M
Jabłoński
Jakub
J
D000328:Adult; D017209:Apoptosis / Q000276:immunology*; D051017:Apoptosis Regulatory Proteins / Q000378:metabolism*; D002294:Carcinoma, Squamous Cell / Q000188:drug therapy / Q000276:immunology* / Q000601:surgery; D053181:Caspase 8 / Q000378:metabolism; D053200:Fas-Associated Death Domain Protein / Q000378:metabolism; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000276:immunology; D008875:Middle Aged; D009062:Mouth Neoplasms / Q000188:drug therapy / Q000276:immunology* / Q000601:surgery; D064549:Myeloid Cell Leukemia Sequence 1 Protein; D009504:Neutrophils / Q000276:immunology*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D053220:Receptors, TNF-Related Apoptosis-Inducing Ligand / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology; D051028:bcl-2-Associated X Protein / Q000378:metabolism
D051017:Apoptosis Regulatory Proteins; C505533:FADD protein, human; D053200:Fas-Associated Death Domain Protein; D064549:Myeloid Cell Leukemia Sequence 1 Protein; D019253:Proto-Oncogene Proteins c-bcl-2; D053220:Receptors, TNF-Related Apoptosis-Inducing Ligand; D051028:bcl-2-Associated X Protein; C505530:CASP8 protein, human; D053181:Caspase 8