Expression And Significance Of HIF-1a,Caspase-3 And Survivin In Esophageal Squamous Cell Carcinoma

[Background and objective]Esophageal carcinoma (EC) is one of the sixth most common malignant turnoutsin the world. Multiple factors and pathways cause the carcinogenesis anddevelopment of the esophageal carcinoma. In recent years, with the development ofmolecular biology, the pathogenesis of the esophageal carcinoma has been knowndeeply, but its true molecular mechanism is not figured out.It has been found that the tumor cells proliferate faster in the stage of no fleshtumor blood vessel, which result in local hypoxia[1].Hypoxia inducible factor 1(HIF-1) is an essential regulatory protein for the adaptation of tumor cells tohypoxia, and regulates cell growth and causes a more malignant phenotype byincreasing the expression of genes encoding angiogenic, metabolic and metastaticfactors. HIF-1 was originally described as a transcription factor that binds to the 3'-enhancer of the erythropoietin (EPO) gene in 1992. Caspase protein is an executeprotein of apoptosis, Caspase—3 is a member of caspase family and it is crucial forprotease responses.The abnormal expression of caspase-3 has been well documentedin a wide variety of malignancies tumors. Survivin protein, a novel anti-apoptosisprotein, is the strongest inhibitor of apoptosis factor to the date. It has been found to beabundantly expressed in a wide variety of human malignancies, while no detected innormal terminally differentiated adult tissues. It has been known that theoverexpressed of survivin in tumor tissues is correlated with poor prognosis of thepatient. This study detects the expression of HIF-1a protein,Caspase-3 protein andSurvivin protein in esophageal squamous cell carcinoma and dysplasia andcorresponding normal epithelium tissues and explore the relation between that and thecarcinogenesis, invasion and lymph node metastasis of esophageal squamous cellcarcinoma by immunohistochemistry. together with the relation of them in apoptosisof esophageal squamous cell carcinoma.This article tries to describe the function ofHIF-1a protein,Caspase-3 protein and Survivin protein and the correlation of them incarcinogensis, development, invasion and metastasis of esophageal squamous cellcarcinoma, in order to find out useful ways to suppress occurrence, development ofesophageal squamous cell carcinoma.[Materials and methods]1. Sample: 49 cases of esophageal squamous cell carcinoma were collected formthe First Affiliated Hospital, Zhengzhou University in 2002. All cases has no thehistory of the chemotherapy, radiotherapy and immunotherapy. All eases wereidentified by pathologist, it is entirely esophageal squamous cell carcinoma. 24 casesareⅠrank.16 cases areⅡrank.9 cases areⅢrank. 23 example appeared the lymphnode metastasis. 26 cases are non-lymph node metastasis. 17 cases are on the low layerinvaded and 32 cases on the deep layer invaded. Each all in site of tumor and tumor-adjacent mucoma cut a piece of tissue involve of 33 cases simple proliferention. 19cases mild dysplasia. 14 cases severe dysplasia. 12 cases carcinoma in situ and 32cases normal esophageal epithelium.2. Reagent:Polyclonal anti-HIF-1a antibody(diluted 1:75) and Polyclonal anti-caspase-3 antibody(diluted 1:100) were both purchased from Boster BiotechCompany, Wuhan. and monoclonal antibody against surviving(diluted 1:30) werefrom Zhonghshan Biotech Company. Beijin. The SP staining kit was bought fromMaixin Company, Fuzhou.3.Method: Using streptomycete avidin peroxides(SP) immunohistochemistry toexamine the expression of HIF-1a protein, Caspase-3 protein and Survivin protein in49 cases esophageal squamous cell carcinoma and precancerous lesions.[Results] 1.The positive rate of HIF-1a protein in esophageal squamous cell carcinoma(51.0％, 25/49) was significantly higher than those in adjacent normalmucosa(0％, 0/32) (P＜0.05). With the increase of invasion depth. the positive rate ofexpression of HIF-1a protein in esophageal squamous cell carcinoma was increased, the positive rates of HIF-1a protein on lower layer invaded of esophageal squamouscell carcinoma group and deeplay invaded of esophageal squamous cell carcinomagroup were 35.3％(6/17) and 65.6％(21/32) respectively, there is a significantdifference between them(P＜0.05).The positive rates of HIF-1a protein in the lymphnode metastasis group and nonlymph node metastasis group were 73.9％(17/23) and38.5％(10/26) respectively, there is a significant difference between them(P＜0.05), too. The positive rates of HIF-1a protein in tumour-adjacent normal epithelium andsimple hyperplasia epithelium, mild atypical hyperplasia epithelium, severe atypicalhyperplasia epithelium, carcinoma in situ and invasive carcinoma were increasedgradually(0％, 30.3％, 31.6％, 35.7％, 41.7％, 51％respectively), asignifieantdifference between the simple hyperplasia epithelium, mild atypical hyperplasiaepithelium, severe atypical hyperplasia epithelium, carcinoma in Situ, invasivecarcinoma and the normal epithelium. While there is no significant difference in thepositive rate of HIF-1a protein in different differentiation carcinoma, although thepositive rate was higher gually(P＞0.05).2. The positive rate of Caspase-3 protein in esophageal squamous cell carcinoma(34.7％, 17/49) was significantly lower than those in adjacent normalmucosa(100％, 32/32) (P＜0.05). The positive rate of Caspase-3 protein in simplehyperplasia epithelium, mild atypical hyperplasia epithelium, severe atypicalhyperplasia epithelium and carcinoma in situ were 75.8％(25/33), 63.2％(12/19), 57.1％(8/14), 66.7％(8/12) respectively, a significant difference between the simplehyperplasia epithelium, mild atypical hyperplasia epithelium, severe atypicalhyperplasia epithelium, carcinoma in situ and the normal epithelium. The positiverates of Caspase-3 protein in the lymph node metastasis group and nonlymph nodemetastasis group were 17.4％(4/23) and 50.0％(13/26) respectively, a significantdifference between them(P＜0.05), the positive rates of Caspase-3 protein on lower layer invaded of esophageal squamous cell carcinoma group and deep layer invadedof esophageal squamous cell carcinoma group were 58.8％(10/17) and 21.9％(7/32)respectively, there is a significant difference between them(P＜0.05), too. While thereis no significance difference in the expression of Caspase-3 protein in differentdifferentiation carcinoma, although the positive rate was lower gually(P＞0.05)3.The positive rates of survivin protein in 32 cases normal esophageal epithelium, 33 cases proliferention, 19 cases mild dysplasia, 14 cases severe dysplasia, 12 casescarcinoma in situ and 49 cases esophageal squamous cell carcinoma were6.3％(2/32), 9.1％(3/33), 31.6％(6/19), 50.0％(7/14), 58.3％(7/12)and 57.1％(28/49) respectively.There is a significant difference between the mild atypicalhyperplasia epithelium, severe atypical hyperplasia epithelium, carcinoma in situ, invasive carcinoma and the normal epithelium (P＜0.05). There is a significantdifference on lower layer invade of esophageal squamous cell carcinoma group anddeep layer invade of esophageal squamous cell carcinoma group together with lymphnode metastasis group and nonlymph node metastasis group (P＜0.05). There is nosignificant difference in different differentiation grades carcinoma tissues (P＞0.05).4.There was a negative correlation between the positive expression rate of HIF-1aprotein and that of caspase-3 protein in esophageal squamous cell carcinoma, therewas 22 cases negative expression of caspase-3 protein in 27 cases positive expressionof HIF-1a protein, there was12 cases positive expression of caspase-3 protein in 22cases negative expression of HIF-1a protein.5. The positive expression of HIF-1a protein was positively correlated with thatof survivin protein in esophageal squamous cell carcinoma, there was 20 casespositive expression of survivin protein in 27 cases positive expression of HIF-1aprotein, there was 14 cases negative expression of survivin protein in 22 casesnegative expression of HIF-1a protein.6. There was a negative correlation between the positive expression rate ofcaspase-3 protein and survivin protein in esophageal squarnous cell carcinoma, therewas 11 cases negative expression of survivin protein in 17 cases positive expressionof caspase-3 protein, there was 22 cases positive expression of survivin protein in 32 cases negative expression of caspase-3 protein.[Conclusion]1. The positive expression of HIF-1a protein in tumor-adjacent atypicalhyperplasia epithilum had a trend of increasing gradually, it hints that over expressionof HIF-1a protein maybe play an important role in the tumorigenesis of esophagealsquamous cell carcinoma.The expression of HIF-1a protein was related to invasionand lymph node metastasis.It was indicated that HIF-1a protein was correlated withinvasion and lymph node metastasis of esophageal squamous cell carcinoma.2. With the lower differented carcinoma tissues, the positive rate of HIF-1a proteinhad a trend of increasing gradually, it hints that HIF-1a protein maybe correlated withthe development of esophageal squamous cell carcinoma.3. Loss or decreased expression of caspase-3 protein is correlated with theoccurence, invasion and metastasis of esophageal squamous cell carcinoma.4. Survivin protein was correlated with the occurrence, invasion and metastasisof esophageal squamous cell carcinoma, .5. There was a negative correlation between the expression of HIF-1a protein andCaspase-3 protein, a positive correlation between the expression of HIF-1a protein andSurvivn protein, it implied that the positive expression of HIF-1a protein candecrease the expression of caspase-3 protein, accordingly, ascending the expression ofsurvivin protein, lead to the inhibition of celluar apoptosis of esophageal squamouscell carcinoma. United examining HIF-1αprotein, Caspase-3 protein and Survivinprotein can become a objective target to evaluate the prognosis of esophagealsquamous cell carcinoma. It had a great significance to judge the prognosis ofesophageal squamous cell carcinoma.