Epidermal growth factor receptor expression in gastric tumors and its relationship with the germline polymorphisms - 216 G>T, -191 C>A, (CA) n IVS1, and R521K.

BACKGROUND: Gastric cancer (GC) is the third worldwide leading cause of cancer-related death affecting both sexes. The aberrant expression of epidermal growth factor receptor (EGFR) gene has been detected in many human epithelial malignancies and linked to advanced disease, more aggressive phenotype, and poor prognosis. AIMS: To analyze the relation that the expression of EGFR in gastric tumors holds with pathological characteristics and with the germline polymorphisms -216 G>T, -191 C>A, (CA) n IVS1, and R521K. MATERIALS AND METHODS: We studied 22 biopsies from gastric tumors obtained by endoscopy. EGFR expression was determined by relative quantification real-time polymerase chain reaction with the glyceraldehyde-3-phosphate dehydrogenase reference gene (as for messenger RNA [mRNA]) and by immunohistochemistry (IHC) (as for protein). EGFR germline polymorphisms were analyzed by sequencing, GeneScan, and restriction fragment length polymorphisms. RESULTS: EGFR mRNA expression was increased (>2-fold) in 13.6% of GC cases, decreased (<0.5-fold) in 68.2%, and normal in 18.2%; overexpression was related to well-differentiated gastric tumors, whereas underexpression was linked to moderate or poorly differentiated gastric tumors (P < 0.001). EGFR protein expression was high (IHC 2+ and 3+) in 29.4% of gastric tumors and was normal or low (score 0 to 1+) in 70.6% cases. EGFR expression, in both mRNA and protein, was not related to any EGFR polymorphism (P > 0.05). CONCLUSIONS: Most gastric tumors showed low EGFR expression (mRNA and protein), whereas EGFR overexpression was related to well-differentiated gastric tumors. Furthermore, germinal polymorphisms -216, -191, (CA) n IVS1, and R521K were not related to EGFR expression (mRNA or protein).

RUNX3 gene polymorphisms and haplotypes in Mexican patients with colorectal cancer.

We analyzed a possible association between RUNX3 gene polymorphisms and haplotypes in Mexican patients with colorectal cancer (CRC). Genomic DNA samples were obtained from the peripheral blood of 176 Mexican patients with CRC at diagnosis and from 195 individuals that formed the control group. The polymorphisms were detected by polymerase chain reaction-restriction fragment length polymorphism. Association was estimated by odds ratio (OR). The haplotypes and linkage disequilibrium were established using the Arlequin v3.5 software. We found that the RUNX3 polymorphisms analyzed were in Hardy-Weinberg equilibrium. The RUNX3 rs2236852 AA genotype and A allele showed association with CRC (OR = 0.39, 95%CI = 0.21-0.73, P < 0.01; OR = 0.65, 95%CI = 0.49-0.87, P < 0.01, respectively), while the rs6672420, rs11249206, and rs760805 polymorphisms did not show significant association with CRC. The TA haplotype (SNPs rs760805 and rs2236852) showed an increased risk for CRC (OR = 2.52, 95%CI = 1.47-4.30, P < 0.001). In conclusion, we found that the AA genotype and A allele of rs2236852 polymorphism confer a decreased CRC risk, while the TA haplotype appears to increase the risk of CRC development in Mexican patients.

Effect of ZNF217 gene polymorphisms on colorectal cancer development in a Mexican population.

The ZNF217 gene, a potential oncogene amplified and overexpressed in several cancers including colorectal cancer (CRC), acts as a transcription factor that activates or represses target genes. The polymorphisms rs16998248 (T>A) and rs35720349 (C>T) in coronary artery disease have been associated with reduced expression of ZNF217. In this study, we analyzed the 2 polymorphisms in Mexican patients with CRC. Genotyping of rs16998248 and rs35720349 sites was performed by polymerase chain reaction-restriction fragment length polymorphism in 203 Mexican Mestizos, 101 CRC patients, and 102 healthy blood donors. Although no statistical differences regarding genotype and allele frequencies of ZNF217 polymorphisms were observed (P > 0.05), linkage disequilibrium was significant in CRC patients (r(2) = 0.39, P < 0.0001), as a result of reduced AC haplotype frequency. Thus, the AC haplotype may protect against CRC.

MDR1 C3435T polymorphism in Mexican patients with breast cancer.

We investigated whether the MDR1 C3435T polymorphism is associated with fibrocystic changes (FCC), infiltrating ductal breast cancer (IDBC), and/or clinical-pathological features of IDBC in Mexican patients. Samples from women who received surgical treatment in 2007 at the Centro Médico de Occidente (México) were included in the analysis. Genotyping was performed by polymerase chain reaction-restricted fragment length polymorphisms in 64 paraffin-embedded breast samples with IDBC, 64 samples with FCC, and 183 peripheral blood samples of healthy females designated as the healthy group (HG). The frequency of the T allele was 41, 45, and 52% for the FCC, IDBC, and HG samples, respectively. Significant differences were only found between the FCC and HG samples [odds ratio (OR) = 0.64, 95% confidence interval (CI) = 0.43-0.96; P = 0.032]. The prevalence of the T/T genotype was 8, 13, and 24% for FCC, IDBC, and HG samples, respectively. Again, statistical differences were only found between FCC and HG samples for the T/T genotype (OR = 0.28, 95%CI = 0.106-0.77; P = 0.009). Although the T allele and the T/T genotype were less frequent in the IDBC group than in the HG, the differences were not significant. Furthermore, no associations were found between the C3435T polymorphism and clinical-pathological features of the IDBC group. Both the FCC and IDBC groups had a high frequency of the C allele relative to the HG in this sample of women from Western Mexico.

Association of MMP7-181A/G and MMP13-77A/G polymorphisms with colorectal cancer in a Mexican population.

Colorectal cancer (CRC) is characterized by enhanced expression and activity of several metalloproteinases (MMPs), including MMP13 and MMP7, which play an important role in tumor invasion and metastasis. The objective of this study was to analyze the association of functional MMP7-181A/G and MMP13-77A/G promoter polymorphisms with susceptibility to CRC in a Mexican population. Genomic DNA samples were obtained from peripheral blood of 102 CRC patients and 125 blood donors who were included as the control group. Identification of polymorphisms was based on polymerase chain reaction-restriction fragment length polymorphism methodology. The association was estimated by the odds ratio (OR) test. The results showed that MMP7-181A/G and MMP13-77A/G variants were associated with CRC. For MMP7-181A/G, the AA (P=0.02, OR=3.38, 95% confidence interval (CI)=1.16-9.84) and AG (P=0.01, OR=3.4, 95%CI=1.17-9.83) genotypes were associated with an increased risk of CRC. For MMP13-77A/G, the AA and AG genotypes were associated with CRC (AA genotype: P=0.04, OR=3.2, 95%CI=1.004-10.2; AG genotype: P=0.01, OR=4.08, 95%CI=1.3-13.07). In conclusion, AA and AG genotype carriers for both polymorphisms are at a higher risk of developing CRC in this Mexican population.

MLH1 and XRCC1 polymorphisms in Mexican patients with colorectal cancer.

DNA repair proteins maintain DNA integrity; polymorphisms in genes coding for these proteins can increase susceptibility to colorectal cancer (CRC) development. We analyzed a possible association of MLH1 -93G>A and 655A>G and XRCC1 Arg194Trp and Arg399Gln polymorphisms with CRC in Mexican patients. Genomic DNA samples were obtained from peripheral blood of 108 individuals with CRC (study group) at diagnosis and 120 blood donors (control group) from Western Mexico; both groups were mestizos. The polymorphisms were detected by PCR-RFLP. Association was estimated by calculating the odds ratio (OR). We found that the MLH1 and XRCC1 polymorphisms were in Hardy- Weinberg equilibrium. The MLH1 655A>G polymorphism in the 655G allele was associated with a 2-fold increase risk for CRC (OR = 2.04 and 95% confidence interval (95%CI) = 1.12-3.69; P < 0.01), while the MLH1 -93G>A polymorphism allele was associated with a protective effect (OR = 0.60, 95%CI = 0.40-0.89; P = 0.01 in the -93A allele and OR = 0.32, 95%CI = 0.13-0.79; P = 0.01 in the AA genotype). The XRCC1 Arg194Trp and Arg399Gln polymorphisms did not show any significant associations. In conclusion, we found that MLH1 -93G>A and 655A>G polymorphisms are associated with CRC in Mexican patients.

Immunological study of a chitosan prosthesis in the sciatic nerve regeneration of the axotomized dog.

This study demonstrated that when the regeneration of the axotomized sciatic nerve is induced through tubulization with chitosan, this biomaterial does not induce immunostimulation or immunodepression in the dog. Canine females were distributed among three groups: an intact control group which was only isolated, an axotomized control group, and an axotomized group which was tubulized with 3% chitosan prostheses. In vitro culture and phagocytosis tests, as well as IgG and IgM serum concentrations, were determined in peripheral blood on days 0, 15, 30 and 60. The results showed that chitosan implants did not importantly affect the immune response.

[Regeneration of the axotomised sciatic nerve in dogs using the tubulisation technique with Chitosan biomaterial preloaded with progesterone]. / Regeneracion del nervio ciatico axotomizado del perro mediante la técnica de tubulizacion con el biomaterial quitosana precargado con progesterona.

INTRODUCTION: Injuries to peripheral nerves can have different causes and may lead to disorders affecting mobility, sensitivity and loss of motor function as they progress. Weiss, in 1944, introduced tubulisation to promote the regeneration of a sectioned nerve. In this study the biomaterial Chitosan was used to induce and stimulate the regeneration of the sciatic nerve in dogs. At the same time, we took advantage of the characteristics offered by Chitosan to include the neurosteroid progesterone in its matrix, as a promoter of axonal growth. AIMS. The aim of our study was to determine the degree of regeneration of the sciatic nerve in dogs when axotomised tubulised with a Chitosan prosthesis steeped in the neurosteroid progesterone. MATERIALS AND METHODS: Young adult female dogs were used to evaluate the regeneration of the sciatic nerve induced at a standard of 15 mm; regeneration was determined by means of an axonal growth chamber. Nerve growth was studied through histological analysis and by electron microscope. RESULTS: The statistical analysis showed that there were no significant differences in the number of myelinated fibres between the experimental groups. The electron microscope images of the transmission in the regenerated nerves in the groups that were tubulised with Chitosan, with and without neurosteroid preloading, revealed an advanced regenerative process. This was evidenced by the fact that collagen fibres, elastin, Schwann cells and both myelinated and non myelinated fibres were observed in all cases. CONCLUSIONS: The regeneration of axotomised, tubulised nerves was achieved regardless of the treatment that was applied. The distal nerve segment that was analysed revealed a similar structure to that of a normal nerve.

Regeneración del nervio ciático axotomizado del perro mediante la técnica de tubulización con el biomaterial quitosana precargado con progesterona / Regeneration of the axotomixed sciatic nerve in dogs using the tubulisation technique with chitosan biomaterial preloaded with progesterone

Introducción. Los nervios periféricos se pueden lesionar por distintas causas, y causar en el curso de su evolución trastornos de movilidad, sensibilidad y pérdida de la función motora. Weiss, en 1944, introdujo la tubulización para promover la regeneración de un nervio seccionado. En este estudio se utilizó el biomaterial quitosana para inducir y estimular la regeneración del nervio ciático del perro, además de aprovechar la cualidad que presenta la quitosana para incluir en su matriz el neuroesteroide progesterona, como un promotor de crecimiento axónico. Objetivo. Determinar el grado de regeneración del nervio ciático del perro axotomizado-tubulizado con una prótesis de quitosana impregnada del neuroesteroide progesterona. Material y métodos. Se emplearon perros hembra adultos jóvenes para evaluar la regeneración del nervio ciático inducida a un defecto de 15 mm, y se determinó su regeneración a través de la cámara de crecimiento axónico. Los crecimientos nerviosos se estudiaron mediante un análisis histológico y por microscopía electrónica. Resultados. El análisis estadístico indicó que no hubo diferencias significativas en el número de fibras mielinizadas entre los grupos experimentales. La microscopía electrónica de transmisión de los nervios regenerados de los grupos tubulizados con quitosana, con y sin neuroesteroide precargado, indicaron un proceso regenerativo avanzado, puesto que en todos se apreciaron fibras de colágeno, elastina, células de Schwann y fibras mielinizadas y no mielinizadas. Conclusiones. La regeneración de los nervios axotomizados y tubulizados se logró con independencia del tratamiento aplicado. El segmento nervioso distal que se analizó evidenció una estructura similar a la de un nervio normal (AU)