| IntroductionColon cancer is the most common gastrointestinal tumor malignancy. In recent years the death rate gradually increased, leading cause of death is caused by tumor invasion and metastasis. In recent years, experimental and clinical data indicate that heparanase HPA in the colon, stomach, liver, esophageal and other gastrointestinal tumors with high expression of invasion and metastasis in some malignant tumors expressed a higher intensity. The expression level and patient prognosis are closely associated. The expression level and patient prognosis are closely associated.S100 protein family is a class of EF-hand type calcium binding protein, through the regulation of calcium and interaction with the target protein, play an important biological role in cell cycle activity, cell differentiation, tumor growth and secretion of extracellular matrix activities, processes, and many members of its abnormal expression in tumors, especially in the abnormal expression of S100A4 with tumor invasion and metastasis. S100 family, and therefore the development of tumors close.This study is to determine the normal colon cancer tissues and the expression of S100A4 in the HPA and discussed both in the strap of the relevance of colon cancer, colon cancer patients for the diagnosis and prognosis of scientific and effective evaluation.Materials and Methods1. Patients and specimens70 cases of colon cancer specimens from 2007 to 2009 at the First Affiliated Hospital of China Medical University, underwent surgical resection of colon cancer specimens. In these 70 specimens were taken in the ends corresponding normal tissue, as no cancer were compared. Fresh specimens from 20 patients with colon cancer from March 2009 to December 2009 at the First Hospital of China Medical University, surgical removal of the fresh specimens. All specimens were obtained cancer, adjacent (within cancer tissues around the 3CM), normal tissue (tumor tissue around the outside 3CM) as control.2. ImmunofluenceRabbit polyclonal anti-human heparanase antibodies were purchased from Santa Inc., rabbit polyclonal anti-human S100A4 antibody was purchased from the Shan Golden Bridge Corporation, SP immunohistochemistry kit, DAB chromogenic enzyme substrate kit was purchased from Fuzhou, the taking of new bio-technology development company. Staining according to kit instructions.3. Western BlotThe organization with 80μg total protein cleavage products were 10% SDS-PAGE electrophoresis 1.5h, the transfer to PVDF membrane (65V,2h), nonfat dry milk closed at room temperature 2h, washed three times by TTBS, anti-HPA polyclonal antibody (1 300) 4℃overnight incubation, the horseradish peroxidase labeled secondary antibody at 37℃for 2h, TTBS washing three times ECL light. Toβ-actin as internal control, to P-actin as internal control, compared with a gray value of each.4. Statistical analysisAll statistical calculations were performed by SPSS version 13.0 for Windows software. p values less than 0.01 were considered statistically significant.ResultsHeparanase HPA and S100A4 in human colon cancer positive rate was significantly higher than normal colon tissue. Both the patients and the sex, age, histological type, differentiation, there is no significant difference between. With tumor invasion and lymph node metastasis are closely related. And positively correlated between the two.ConclusionsIn colon adenocarcinoma, heparanase HPA and S100A4 protein expression is closely related with tumor progression, is predicting the biological behavior of the transfer trend forecasts valuable indicators. Combined detection of heparanase HPA and S100A4 protein expression can be used to assess colon cancer metastasis.
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