6th ICHNO Abstract Book

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page 48 6th ICHNO
6 th ICHNO Conference
International Conference on innovative approaches in Head and Neck Oncology
16 – 18 March 2017
Barcelona, Spain
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patients about the important implications of HPV
positivity.
PO-100 Oral cancer microenvironment -
PI3K/AKT/mTOR and ERK/MAPK pathways dependent
targets
C. Domingues1,2,3, M. Laranjo3,4, A.M. Abrantes3,4, A.B.
Sarmento-Ribeiro2 , L. Carvalho 3,5, M.F. Botelho3,4, H.
Silva3,6, M. Dourado1,2,3
1Faculty of Medicine- University of Coimbra- Portugal,
Pathophysiology Curricular Unit- Dental Medicine,
Coimbra, Portugal
2Faculty of Medicine- University of Coimbra- Portugal,
Laboratory of Oncobiology and Hematology- Applied
Molecular Biology and University Clinic of Hematology,
Coimbra, Portugal
3Faculty of Medicine- University of Coimbra- Portugal,
Center of Investigation on Environment Genetics and
Oncobiology CIMAGO, Coimbra, Portugal
4Faculty of Medicine- University of Coimbra- Portugal,
Biophysics Institute- Institute for Biomedical Imaging and
Life Sciences IBILI, Coimbra, Portugal
5Faculty of Medicine- University of Coimbra- Portugal,
Institute of Anatomical and Molecular Pathology,
Coimbra, Portugal
6Faculty of Medicine- University of Coimbra- Portugal,
Medical Genetics Institute, Coimbra, Portugal
Purpose or Objective
Oral cancer represents 30% of all head and neck cancer
and over 95 % are squamous cell carcinomas (OSCC). The
anatomy of the oral cavity is particularly challenging due
to different microenvironments identified in this relatively
small area. Thus, the interplay between tumor cells and
stromal structure has been receiving growing attention as
it can contribute for malignant transformation. The
PI3K/AKT/mTOR and the ERK/MAPK pathways regulate
cell survival, proliferation, and motility and can converge
to regulate each other.
The aim of our study is to understand the role of
PI3K/AKT/mTOR and ERK/MAPK in oral cancer
microenvironment, namely in the reciprocal interplay
between malignant cells and stromal structure, in order to
identify new diagnosis and therapeutic targets for OSCC.
Material and Methods
For this purpose, the human OSCC cell line, HSC-3, was
cultured. Then, approximately 3, 15 or 30 million were
inoculated in Balb/c nu/nu mice (4/location) on tongue,
on jugal submucosa and on dorsal side subcutaneously
(back), respectively (Of. 3-CE-2011 FMUC, Portugal).
During 3-7 weeks, the mice were supervised. After, they
were killed by anesthetic overdose and tumor nodules and
involving tissue were excised for ex vivo studies. The
histology was assessed by the hematoxylin-eosin staining.
The immunophenotype was evaluated by
immunohistochemistry (IHC) using AE1AE3 cytokeratin
cocktail, Vimentin, phospho(p)mTOR and Ki67. For genetic
studies the samples were Purezol® processed and the
obtained RNA was converted to cDNA and a 48 RT-PCR
array for PI3K/AKT/mTOR pathway specific for human was
applied.
Results
Our results showed that depending on the
inoculation/implantation site HSC-3 cells exhibit different
gene expression and immunophenotype profile as well as
different amount of keratinizing cells, large cells and
basal/intermediate cells. Tongue location showed
increased IHC expression of AE1/AE3, Vimentin and
pmTOR comparing with the other models. The Ki67 index
was higher in large and basal cells in all models. The gene
expression profile showed that the PI3K/AKT/mTOR
pathway has a different transcription regulation according
to the tumor implantation site. The jugal submucosa
location showed increased expression of AKT3, PDK1,
PDK2, PRKCA, IGF1R, MAPK3/ERK1, CTNNB1 (1.5x higher),
and decreased expression of FASL and NFKB1A (0.5x
higher). These results with IHC unstaining of pmTOR
indicate that in jugal submucosa MAPK/ERK pathway has a
critical role in carcinogenesis.
Conclusion
Our preliminary results suggest that tumor
microenvironment determines different gene expression
and immunophenotypic profile in OSCC. In tongue the
PI3K/AKT/mTOR pathway appears to have a determinant
role whereas in jugal submucosa the ERK/MAPK pathway
maybe a preferential target.
PO-101 Osteopontin (OPN) as an instant, predictive
marker of tumour hypoxia in head and neck cancer
patients
J. Mrochem-Kwarciak 1, T. Rutkowski2, A. Wygoda2, R.
Deja1, A. Hajduk2, Ł. Boguszewicz3, P. Widłak4, K.
Składowski2
1Maria Sklodowska-Curie Memorial Cancer Center and
Institute of Oncology, ANALYTICS AND CLINICAL
BIOCHEMISTRY DEPARTMENT, Gliwice, Poland
2Maria Sklodowska-Curie Memorial Cancer Center and
Institute of Oncology, I Radiation and Clinical Oncology
Department, Gliwice, Poland
3Maria Sklodowska-Curie Memorial Cancer Center and
Institute of Oncology, Department of Medical Physics,
Gliwice, Poland
4Maria Sklodowska-Curie Memorial Cancer Center and
Institute of Oncology, Center for Translational Research
and Molecular Biology of Cancer, Gliwice, Poland
Purpose or Objective
Despite of well known prognostic factors like locoregional
stage of disease, there is still lack of markers describing
individual feature of the tumor. Some of proinflammatory
cytokines are supposed to predict the nature of the tumor
what may help to optimize the therapy. Plasma
osteopontin (OPN) is a putative marker of tumor hypoxia
in patients with head and neck squamous cell carcinomas
(HNSCC). Additionally, OPN has been recognized as
important in the processes of tumorigenicity and
metastasis. The aim of the study was to assess clinical
utility of OPN as the biomarker of treatment outcome of
radiotherapy (RT) or radiochemotherapy (CHRT) in HNSCC.
Material and Methods
Between 01/2009 and 08/2013 251 patients in the mean
age of 59 years with squamous cell carcinoma of
oropharynx (39%), hypopharynx (13%), larynx (44%) or oral
cavity (4%) were treated with RT alone (48%) or combined
with chemotherapy (52%). The median duration of
symptoms prior the treatment was 33 months (range: 1 –
70). The stage of disease was determined due to a TNM
scale. There were 15 (6%), 112 (45%), 74 (29%), and 50
(20%) patients with T1, T2, T3 and T4 tumor stage,
respectively, and 99 (40%), 26 (10%), 105 (42%), and 21
(8%) patients with N0, N1, N2 and N3 nodal stage of
disease, respectively (no patients with distant metastases
were included). OPN was indicated in plasma before
treatment and immediately after treatment completion.
In statistic overview of the results STATISTICA 9.1
(StatSoft) program was used. While interpreting the
results median value was used. U Mann-Whitney test was
used for analysis of correlation between protein
concentration and the stage of the disease. Multivariate
Cox analysis of factors related to OS was carried on. Log-
rank test was used to compare OPN as categorized value
acc. to median respectively.
Results