Research Of Screening Different Protein In Esophageal Squamous Cell Carcinoma And Clinical Significance

Objective: To explore research of screening different protein in esophageal squamous cell carcinoma and clinical significance. Methods: 1): Laser capture microdissection was used to isolate epithelial cells from adjacent inflammatory and cancer cells. 2): An obviously different expressed protein spot was separated by two dimensional electrophoresis(2-DE), then analyzed and identified as different proteins using Matrix Assisted Laser Desorption Ionization Time of Flight Mass spectrometry(MADLDI-TOF-MS). 3): Western blot and Immunohistochemistry technique was performed to detect the expression of Bmi-1, PAI-1, Cofilin-1 and Transgelin in esophageal squamous cell carcinoma tissue. 4): The relationships between the expression Bmi-1, PAI-1, Cofilin-1, Transgelin, and the clinicopathologic features of ESCC, and the survival rate of ESCC patients were also discussed. Results: 1): LCM is powerful to obtain a pure targeted cell subgroup from ESCC and Normal esophageal tissue, quickly and precisely under the microscope. 2): It identified 43 significant differences proteins by 2-DE combined MADLDI-TOF-MS. There are 25 high expression proteins in ESCC of Kazak and Han. There are 14 down regulation expression proteins in ESCC of Kazak and Han.There are four differentially expressed proteins in ESCC of Kazak and Han. 3): The expression of Bmi-1, PAI-1, Cofilin-1 and Transgelin in ESCC is significantly higher than the normal tissues by western blot.The rates of expression of Bmi-1, PAI-1, Cofilin-1 and Transgelin in esophageal cancer tissue were 78.75%(P<0.05), 76.25%(P<0.05), 60.00%(P<0.05), 55.00%(P<0.05), respectively. It was higher than that in normal esophageal tissues. 4): Analysis correlation with gender, age, nationality, tumor size, degree of differentiation, pathologic classification, clinical stage, invasion depth, lymph node metastasis of Bmi-1, PAI-1, Cofilin-1 and Transgelin in ESCC. The rates of expression of Bmi-1 with lymph node metastasis and without lymph node metastasis were 63.0 and 86.6%, respectively. Bmi-1 protein expression in patients with lymph node metastasis was lower than that in patients without lymph node metastasis, and the difference was statistically significant(P<0.05). Bmi-1 expression gradually increased with increased depth of infiltration in the muscularis and serosa layer, and the difference was statistically significant(P<0.05). The rates of expression of PAI-1 with lymph node metastasis and without lymph node metastasis were 59.3% and 84.9%, respectively.PAI-1 protein expression in patients with lymph node metastasis was lower than that in patients without lymph node metastasis, and the difference was statistically significant(P<0.05). PAI-1 expression gradually increased with increased depth of infiltration in the muscularis and serosa layer, and the difference was statistically significant(P<0.05). The rates of expression of Cofilin-1 with lymph node metastasis and without lymph node metastasis were 88.9% and 45.3%, respectively, and the difference was statistically significant(P<0.05). The differences of Cofilin-1 positive expression rates in patients with high, moderate, or low differentiation were 26.1%, 78.3% and 70.6%, respectively, and difference was statistically significant(P<0.05).It indicates that Cofilin-1 expression gradually increased with low degree of differentiation. At the clinical stage I-II and III-IV, the positive expression of Cofilin-1 were 37.1% 和 77.8%, respectively, and the differences were statistically significant(P < 0.05). The results indicated that the expressions of Cofilin-1 probably were close related to clinical stages.Cofilin-1 expression in ESCC was not correlativity with sex, age, nationality, tumor size and depth of infiltration. There was no significant difference in expression(P > 0.05). The Transgelin positive expression rates in patients with high, moderate, or low differentiation were 34.8%、52.2% and 70.6%, respectively, and the difference weas statistically significant(P<0.05). It indicates that Transgelin plays an important role in cell differentiation. Transgelin expression in patients without lymphatic metastasis and with lymphatic metastasis were 37.7% and 88.9%, respectively.The difference was statistically significant(P<0.05), which showed that Transgelin expressions were higher in patients with lymphatic metastasis. The results indicate that Transgelin expression in ESCC had no relation with sex, age, nationality, depth of infiltration, clinical stages and tumor size(P>0.05). Conclusion: 1): Isolation of pure targeted cells in LCM is an important and essential step for the molecular analysis of cells involved in ESCC. 2): The occurrence and development mechanism of ESCC depends on alteration of multi-factors, multi-stages and multi-genes, involving many kinds of oncogene activation and anti-oncogene inactivation, a variety of protein function the result of the collaborative work together. 3): The results of Western blot and Immunohistochemistry of Bmi-1, PAI-1, Cofilin-1 and Transgelin is in accord with proteomics. 4): Up-regulation of Bmi-1, PAI-1, Cofilin-1 and Transgelin protein expression may play an important role in the tumorigenesis and development of ESCC. They may be markers of the growth and metastasis of ESCC. This study will play a role in the research and development of molecularly targeted therapy, and provide a reliable theory for ESCC gene therapy.