Sabato 27 ottobre 2012 - Pacini Editore

PoStER
for protein expression of PAR-2 and density of MC-Try by
immunohistochemical double-staining. PAR-2 expression was
evaluated both in the areas overexpressing the receptor and at the
sites where MC most intensively accumulated. The relationship
of these two tumor markers with clinicopathologic parameters
were also analyzed.
Results.The MC count in normal mucosa adjacent to colon cancer
(79.2 MC/mm 2 , range 22.5-192.7) was significantly higher than
density in the stroma of the primary CRC (56.3 MC/mm 2 , 15.8-
97.8) (p < 0.000). Tumor invasive front showed a higher PAR-2
expression (26.5%, 6.7-62.0) than expression in normal mucosa
(2.70%, 0-17.7) (p < 0.000). Also in areas where MC-Try most
intensively accumulated, tumor showed a statistically higher median
expression of PAR-2 (34.4%, 7.7-78.4) than normal mucosa
(2.5%, 0-18.9) (p < 0.000). In tumor compartment, a significant
positive correlation was found between PAR-2 expression and
MC-Try density (r = 0.393, p< 0.018).
Moreover, it was demonstrated a higher PAR-2 expression in
tumors with poor differentiation grade (p = 0.005), positive Peritumoral-Vascular–Invasion
(p = 0.010) and with positive lymph
node (p = 0.007) and distant metastasis (p = 0.002) status. Additionally,
higher MC-Try density occurred in poor differentiation
grade cancers (p = 0.031) and in male patients (p = 0.004).
Conclusions. We found that the distribution of MC and PAR-2
varied passing from tumor to adjacent normal mucosa and the
increased expression of PAR-2 was positively related to MC-Try
density and to colonic tumors with poor prognosis. The results
of our study suggest that, in the context of developing CRC,
infiltrating MC-Try might influence the expression of PAR-2,
contributing to tumor invasion and metastasis. Further studies are
needed to support these observations.
references
1 Maltby S, Khazaie K, McNagny KM. Mast cells in tumor growth:
angiogenesis, tissue remodelling and immune-modulation. Biochim
Biophys Acta 2009;1796:19-26.
2 Rattenholl A, Steinhoff M. Proteinase-activated receptor-2 in the
skin: receptor expression, activation and function during health and
disease. Drug News Perspect. 2008;21:369-81.
3 Nishibori M, Mori S, Takahashi HK. Physiology and pathophysiology
of proteinase-activated receptors (PARs): PAR-2-mediated proliferation
of colon cancer cell. J Pharmacol Sci 2005;97:25-30.
An association study of two single nucleotide
polymorphisms (SnPs) in the SPArC and EGF genes
with hepatocellular carcinoma
S. Marasà1 , D. Balasus2 , A. Giacalone2 , L. Marasà2 , L. Giannitrapani2
, E. Sinagra3 , G. Montalto2 1 2 Institute of Pathologic Anatomy, University of Palermo; Department
of Clinical Medicine and Emerging Pathologies, University of Palermo;
3 Division of Internal Medicine, V.Cervello Hospital, Palermo
Introduction. Hepatocellular carcinoma (HCC) is responsible
for 85% of all the liver tumors 1 . It is the sixth most common
neoplasm and the third most frequent cause of cancer-related
deaths worldwide 2 . HCC incidence varies among countries and
continents and, is strictly connected to endemic risk factors.
Theoretically any agents leading to chronic liver inflammation
could be risk factors. In fact HCV (hepatitis C virus) and HBV
(hepatitis B virus) infections are widely recognized HCC risk
factors, they account for 70-80% of HCC cases. The remaining
percentage of HCCs is caused primarily by aflatoxin B, alcohol,
hemochromatosis and autoimmune hepatitis 3 , HCC often develops
from a state of liver cirrhosis.. As for gender, males have
higher liver cancer rates than females with a 3 to 1 ratio 4 . Despite
of the fact that HCV infection is the main HCC risk factor not all
of the HCV-positive subjects develop the disease. This happens
for almost all the others risk factors. In fact many have founded
a lot of genetic variations that predispose to liver cancer onset
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and development 5 . In particular a lot of single nucleotide polymorphisms
(SNPs) have been associated to HCC 6 . Nevertheless
genetic variations may have different involvements in the disease
depending on the ethnic groups. In this context the secreted
protein acidic and rich in cysteine (SPARC) and the epidermal
growth factor (EGF) protein could be involved in hepatocellular
carcinogenesis. The aim of this study was to evaluate the association
between HCC susceptibility and the rs2304052(position
151034420, substitution T > C) and the rs4444903(position
111053559, substitution > A) single nucleotide polymorphisms
(SNPs), located respectively on the SPARC and EGF protein.
Methods and materials. Genomic DNA was extracted from the
whole blood samples (75 HCC cases and 170 healthy controls
were collected from a Southern Italian population).DNA was
isolated from Buffy coat using the High Pure PCR Template
Preparation Kit (Roche®). DNA concentration was evaluated
for every sample through a comparison with four standard DNA
samples having known concentrations. Genotypes were detected
by restriction fragment length polymorphism (RFLP) method.
For the rs2304052 SNP the restriction reaction was performed in
20 µl with 2 U of EcoO109I. The digestion products were electrophoresed
on a 2% agarose gel. The digestion with EcoO109I
produced two fragments of 222 bp and 197 bp in case of C homozygous,
three fragments of 419 bp, 222 bp and 197 bp in case of
heterozygous and one fragment in case of T homozygous as it is
shown in Figure 1. For the rs4444903 SNP the restriction reaction
was performed in 20 µl with 1 U of AluI. The digestion with AluI
produced two fragments of 91 bp and 187 bp in case of A homozygous,
three fragments of 278 bp, 187 bp and 91 bp in case of heterozygous
and one fragment of 278 bp in case of G homozygous
as it is shown in Figure 2. Genotype frequencies were evaluated by
direct gene counting. Control and cases alleles distribution fitted
to Hardy-Weinberg equilibrium (p 2 + 2pq + q 2 = 1, where p is the
Fig. 1. 2% agarose gel stained with SYBR Safe®. lane 1, 2: C/t heterozygous.
lanes 3, 5, and 6: t/t homozygous.lane 4: C/C homozygous.
l: 100 bp DNa ladder.
Fig. 2. 2% agarose gel stained with Gel Red®. lane 1: a/a homozygous.
lane 2: G/a heterozygous. lane 3: G/G homozygous. l: 100 bp dNa ladder.