Expression And Significance Of Cell Cycle Regulatory Factor In Canceration Of Esophageal Epithelium

Objective:To investigate the effects of the gene expressions of cell cycle regulatory factor in normal esophageal mucosal epithelia, intraepithelial neoplasia and esophageal squamous cell carcinom. To discuss the reason of Unbalance of apoptosis regulation. The aim was provide valuable evidence for l diagnosis and genetic of esophageal squamous cell carcinoma. Compare the results of immunohistochemistry method and the results of double-label immunofluorescence method, to study the significance of double-label immunofluorescence method associated with Laser Scanning Confocal Microscopy in medical reseach.Methods:120 esophageal squamous cell carcinoma samples were collected to make tissue microarray. Immunohistochemical method (Envision) was used to detect the protein expression of cell cycle regulatory factor in tissue microarray. The results were statistically analyzed. Using double-label IF, the expression of cell cycle regulatory factor were detected. The results were observed and photographed by Laser Scanning Confocal Microscopy. Compare the results of two methods. Acquired data were analyzed by SPSS13.0 statistical software。Result:(1) The positive ration of cyclinD,CDK4,cyclinE,CDK2 protein expression in normal esophageal epithelium is low (28.3%,28.3%,16.7%,28.3%), it was increased in esophageal intraepithelial neoplasia, and it was high in esophageal squamous cell carcinoma (67.5%,84.2%,46.7%,63.3%). Which was increased with the degree of differentiation decreasing gradually. There was a significant difference among the three groups. The expression of CDK4 in group with lymphatic metastasis was higher than without(P=0.015,<0.05).(2)The positive ration of p14,p15,p18,p19,p21,p27 protein expression in normal esophageal epithelium are high(29.2%,30.0%,34.2%,29.2%,49.2%,45.8%), it were decreased in esophageal intraepithelial neoplasia, but it were high in esophageal cancer which were decreased with the degree of differentiation gradually increased.(3)The expression of p16,p57 in normal esophageal epithelium is higher than in esophageal intraepithelial neoplasia and esophageal squamous cell carcinoma. And it was very low in esophageal squamous cell carcinoma(11.7%,27.5%).(4)In esophageal squamous cell carcinoma, p18 protein expression was correlated with both cyclinD1 and CDK4(r=0.621,0.696), p19 protein expression was correlated with both cyclinD1 and CDK4(r=0.615,0.630); there was significant positive correlation between the protein expression of cyclinD1 and CDK4(r=0.670); there was significant positive correlation between the protein expression of p18 and p19(r=0.833).(5)Compare the results of immunohistochemistry method and the results of double-label immunofluorescence method, we found that both cyclinD1 and p18 protein expression in normal esophageal epithelium, intraepithelial neoplasia and esophageal squamous cell carcinoma.Conclusion:(1)cyclinD1,CDK4,cyclinE,CDK2 in normal esophageal epithelium is expressed low, it was increased in esophageal intraepithelial neoplasia, and it was high in esophageal squamous cell carcinoma. Which was increased with the degree of differentiation decreasing gradually. There were significant differences among the three groups. They play important roles during growth and development of esophageal squamous cell carcinoma. The expression of CDK4 in group with lymphatic metastasis was higher than without. The over- expression of CDK4 contribute to the proliferation of esophageal carcinoma. (2)p14,p15,p18,p19,p21,p27 in normal esophageal epithelium is expressed high. They were low in esophageal intraepithelial neoplasia, but it was high in esophageal cancer which were decreased with the degree of differentiation gradually increased. (3)The expression of p16,p57 in canceration of esophageal epithelium are decreased. (4)In esophageal squamous cell carcinoma, the expression of p18 and cyclinD1 p19 and CDK4 p18 and p19 show significant relation. It indicates that cyclinD1 and CDK4 may induce the expression of p18 and p19. (5)The results of IF method were accordant with immunohistochemistry Two-step method. IF method combined with LSCM can detect the coexpression of two proteins, so they have important significance in medical research.