RESUMO
El cáncer gástrico es una de las patologías malignas más frecuentes en el mundo. En las últimas décadas la atención se ha centrado en posibles alteraciones de factores genéticos que incluyen la activación de proto-oncogenes y/o la inactivación de genes supresores tumorales. El producto del C-MET proto-oncogen y su ligando, el factor de crecimiento del hepatocito (HGF), han sido implicados en la proliferación y migración celular en el cáncer gástrico. En este estudio se analizó a nivel molecular la amplificación del ARNm del receptor c-Met a partir del tejido tumoral gástrico de pacientes a quienes se les practicó gastrectomías, utilizando el método del ácido guanidina-tiocianato-fenol-cloroformo, y el método semicuantitativo de la Reacción en Cadena de la Polimerasa con Transcriptasa Reversa (RT-PCR), encontrándose que los elevados niveles del ARNm del receptor c-Met en las muestras tumorales de los pacientes están relacionados con mayor invasión en la profundidad de la pared gástrica (r = 0,762, p<0,01), incremento en la metástasis a los ganglios linfáticos (r = 0,766, p<0,01), alta frecuencia en tumores pocos diferenciados o indiferenciados (r = 0,912, p<0,001), aumento en el estadiaje del cáncer gástrico (r = 0,838, p<0,001), y en la sobreexpresión por el método inmunohistoquímico (IHQ) de la estreptavidina-biotina marcada de su receptor a nivel proteico (r = 0,858, p<0,001). La amplificación del ARNm y/o la sobreexpresión a nivel proteico del receptor c-Met, pudieran ser utilizados como factores pronósticos en el cáncer gástrico.
Gastric cancer is one of the most common malignancies in the world. In the last decades, the attention has been focused in possible alterations of genetic factors that include proto-oncogene activation and/or the tumor suppressor gene inactivation. The product of the proto-oncogene c-MET and its ligand, hepatocyte growth factor (HGF), have been implicated in cell proliferation and migration in gastric cancer. In this study we analyzed at the molecular level, the amplification of c-Met receptor mRNA from gastric tumor tissue of patients who underwent gastrectomy, using the acid guanidinium-thiocyanate-phenol-chloroform method and the semiquantitative Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR) method. It was found that high levels of c-Met receptor mRNA in tumor samples from patients are associated with greater depth of invasion in the gastric wall (r = 0.762, p<0.01), increase in metastases to lymph nodes (r = 0.766, p<0.01), high frequency of poorly differentiated or undifferentiated tumors (r = 0.912, p<0.001), increase in the gastric cancer staging (r = 0.838, p<0.001), and the overexpression, by the immunohistochemistry method (IHC) of the labeled streptavidin-biotin, of the c-Met receptor at the protein level (r = 0.858, p<0.001). The amplification of mRNA and/or protein level overexpression of the c-Met receptor could be used as prognostic factors in gastric cancer.
Assuntos
Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Carcinoma/genética , Regulação Neoplásica da Expressão Gênica , Proteínas de Neoplasias/biossíntese , Proteínas Proto-Oncogênicas c-met/biossíntese , RNA Mensageiro/biossíntese , RNA Neoplásico/biossíntese , Neoplasias Gástricas/genética , Diferenciação Celular , Estudos Transversais , Carcinoma/metabolismo , Carcinoma/patologia , Carcinoma/cirurgia , Gastrectomia , Metástase Linfática , Invasividade Neoplásica , Estadiamento de Neoplasias , Proteínas de Neoplasias/genética , Proteínas Proto-Oncogênicas c-met/genética , Neoplasias Gástricas/metabolismo , Neoplasias Gástricas/patologia , Neoplasias Gástricas/cirurgiaRESUMO
Cyclin-dependent kinase inhibitors (CDKI) are negative regulators of cell cycle progression by binding the cyclin-CDK complex and inhibiting the CDK activity. Genetic alteration in the CDKI genes has been implicated for carcinogenesis. To test the genetic alteration in the p27 and p57 genes, KIP family CDKI genes, 30 gastric tumor-normal pairs and 8 gastric cancer cell lines were analyzed for mutations by polymerase chain reaction-single strand conformational polymorphism (PCR-SSCP). No mutation was detected in these genes although length polymorphisms in the proline-alanine repeat of the p57 gene were detected. When the p27 and p57 mRNAs were analyzed in gastric cancer cell lines by RT-PCR, the p27 mRNA was expressed considerably high in tumor cells but expression of the p57 mRNA was much low in gastric cancer cell lines compared to that of normal cells. The result suggests that inactivation of gene expression rather than mutations in the p57 gene accounts possibly for the involvement of this gene in tumorigenesis of gastric cancer. However, expression of the p27 gene seems to be essential for cell survival.