| Background and objectiveEsophageal carcinoma is one of the six most common malignant tumor in theworld. The mortality rate of esophageal carcinoma is the top 10 main malignantneoplasms, and about 2 millions of persons died of it in the world each year. In China,Esophgeal carcinoma is the fourth in mortality rate of malignant tumor and is thesecond in mortality rate of female malignant tumor. In China, Taihangshan is a placewith the highest incidence of esophageal carcinoma, and Chaoshan region of easternGuangdong province is also a major region where has a higher incidence andmortality rate. During the process of growth, Cell proliferation and apoptosis keep adynamic balance under rigorous control. Disorder of the balance effects directly thebiological behavior of tissue and results in the occurrence of tumor. More and moreresearches show that excessive proliferation and aberrant apoptosis play an importantrole in the occurrence & development of malignant tumor. Concerning the relationshipbetween apoptosis-regulating genes and esophageal carcinoma, the relationshipbetween apoptosis-regulating genes and ESCC,carcinomas with adenoid structureand small cell carcinoma checked simultaneously have not been published. This studywas designed to investigate the expression of PTEN,MGMT,Survivin and IPO-38 inesophageal carcinoma and paracancerous tissue, using immunohistochemistry(EnVision two-step method)and confocal laser scanning microscope, so to explore therelationship between the above four proteins expression and the carcinogenesis ofesophageal carcinoma and to find biological index for the early diagnosis andprognosis of esophageal carcinoma.Materials and Methods1 Tissue Samples ①85 cases of paraffin biopsy specimens was collected form the department ofpathology of Shantou university medical college during 2001.11～2005.7.There were 27 cases with paracancerous mucosa among the 85 cases ofesophageal carcinoma.②20 cases of fresh biopsy specimens was collected from the department ofpathology of Shantou university medical college during 2004.7～2005.7.Specimens were all fixed in 10% formaldehyde solution and embedded inparaffin.2 HE stain: All the paraffin biopsy specimens were to make tissue morph andpathological diagnosis after HE staining.3 Immunoenzyme immunohistochemistry techniques and lightmicroscope: expression of PTEN,Survivin,IPO-38 and MGMT protein in 85cases of esophageal carcinoma and 27 cases of paracancerous mucosa wasdetected by immunohistochemistry techniques(EnVision two-step method).4 Indirect immunofluorescence techniques and confocal laserscanning microscope: qualitative and quantitive expression of PTEN andMGMT protein in 20 cases of esophageal carcinoma was observed under theconfocal laser scanning microscope after Indirect immunofluorescence staining,and the relationship between the expression and differentiation of the esophagealcarcinoma was considered.Result1. Histopathology: According to the HE staining, the histological types ofesophageal carcinoma were made under the light microscope (WHOclassification).Among the 85 esophageal carcinomas there are 56 ESCC, 9 small cellcarcinomas and 20 carcinomas with adenoid structure. The latter includes 9mucoepidermoid carcinomas, 9 adenosquamous carcinomas and 2 adencarcinomas.Among the 27 paracancerous mucosa, there are 16 hyperplasia and 11 displasia. Invasion depth and the positive rate of the lymph node metastasis compared in theESCC,carcinoma with adenoid structure and small cell carcinoma, Only between theESCC and carcinomas with adenoid structure there was significantdifference,compared the Invasion depth (P＜0.05).The positive rate of lymph nodemetastasis in small cell carcinoma was significantly higher than that in ESCC (P＜0.05);There was no significant difference in carcinomas with adenoid structure andESCC,small cell carcinoma (P＞0.05).2. Immunohistochemistry2.1 The expressions of the above four proteins in different lesions ofesophagealThe expression of PTEN protein was detected in tumor nuclei. PTEN wasparticularly in the basal cell layer. From hyperplasia to displasia to esophagealcarcinoma, the expression of PTEN protein demonstrated descending tendency, andthat in hyperplasia was significantly higher than in displasia and esophagealcarcinoma, but the significant difference was observed in the expression of PTENonly between hyperplasia of esophageal and esophageal carcinoma (P=0.005). Theexpression of MGMT protein was detected in nuclei and cytoplasm or only nuclei.MGMT was particularly in the basal cell layer. Significant difference was observed inthe expression of MGMT between esophageal carcinoma and hyperplasia,displasia ofesophageal (P=0.000, P=0.014). The expression of Survivin protein was detected innuclei and cytoplasm or only nuclei, and that of IPO-38 protein only in nuclei.Survivin and IPO-38 was particularly in the parabasal cell layer. From hyperplasia todisplasia to esophageal carcinoma, the expression of Survivin and IPO-38demonstrated ascending tendency. The expression of Survivin showed significantdifference between hyperplasia and esophageal carcinoma (P=0.020), but There wasno correlation between the expression of IPO-38 and different lesions of esophageal(P＞0.05).2.2 The expression of the four proteins in various types of esophageal carcinomaThe expression of PTEN was significantly lower in the small cell carcinoma(33.3%) than that in the esophageal carcinoma (42.9%) and that in carcinomaswith adenoid structure (75.0%), There was significant difference between ESCC,small cell carcinoma and carcinomas with adenoid structure (P=0.014, P=0.036). Theexpression of MGMT in ESCC and carcinomas with adenoid structure wassignificantly higher than that in small cell carcinoma (P＜0.01). The expression ofSurvivin in small cell carcinoma was higher than that in ESCC and carcinomas withadenoid structure, but the significant difference was observed in the expression ofSurvivin only between small cell carcinoma and carcinomas with adenoid structure(P=0.001). There was no correlation between the expression of IPO-38 andhistological types of esophageal carcinoma.2.3 The relationship between the four proteins expression andclinicopathological character of ESCCFrom the well differentiated ESCC to moderate differentiated ESCC to poordifferentiated ESCC,The expression of PTEN demonstrated descending tendency, Thepositive rate of PTEN expression in well differentiated ESCC was higher than that inmoderate differentiated ESCC (P＜0.05);There was no significant difference betweenthe well differentiated ESCC,moderate differentiated ESCC and poor differentiatedESCC. The expression of Survivin with lymph node metastasis was significantlyhigher than those had not (P＜0.05); The expressions of Survivin had correlation withinvasion depth but not with differentiation. The expressions of MGMT and IPO-38protein had no correlation with differentiation, invasion depth and lymph nodemetastasis of esophageal carcinoma.2.4 The correlationship of the four proteins in esophageal carcinomaIn 85 esophageal carcinomas, Pearson correlation was used to analyze therelationship of the four proteins. It was found that a positive correlation was observedbetween the expressions of PTEN and MGMT(r=0.231, P＜0.05); a positive correlation was observed between the expressions of Survivin and IPO-38(r=0.878, P＜0.01); anegative correlation was observed between the expressions of PTEN andSurvivin(r=-0.235, P＜0.05);but there was no correlation between the expression ofPTEN and IPO-38(r=0.185, P＞0.05).3 Indirect immunofluorescence techniques and confocal laserscanning microscopeThe expression of MGMT protein was detected in nuclei and cytoplasm or onlynuclei. The expression of PTEN protein was detected massively or lamellarly only innuclei. IOD represents the expression of it when the expression PTEN and MGMTprotein was investigated by confocal laser scanning microscope. The quantity of PTENexpression in well differentiated ESCC (88195.69±50794.95) was higher than that inmoderate differentiated ESCC (28100.27±35988.47), t=3.053, P＜0.01. The quantityof MGMT expression in well differentiated ESCC (12782.23±19465.57)was lower thanthat in moderate differentiated ESCC (19961.17±24151.26), but there was nosignificant difference between the well differentiated ESCC and the moderatedifferentiated ESCC.Conclusion1. From hyperplasia to displasia to esophageal carcinoma, the expression of PTENprotein demonstrated descending tendency and that of Survivin protein demonstratedascending tendency. Significant difference was observed in the expression of MGMTbetween esophageal carcinoma and hyperplasia,displasia of esophageal (P=0.000,P=0.014). These data indicated that the abnormal expression of PTEN,Survivin andMGMT correlates with carcinogenesis of esophageal carcinoma. It may be an earlymolecular event in the development of esophageal carcinoma.2. The positive rate of PTEN expression in well differentiated esophageal squamouscell carcinoma was higher than that in moderate differentiated esophageal squamouscell carcinoma(P＜0.05);The expression of Survivin with lymph node metastasis wassignificantly higher than those had not(P＜0.05), Which indicated that expression of PTEN and Survivin is related to the differentiation and prognosis of esophagealcarcinoma.3. A positive correlation was observed between the expressions of PTEN andMGMT(r=0.231, P＜0.05); A positive correlation was observed between theexpressions of Survivin and IPO-38(r=0.878, P＜0.01); A negative correlation wasobserved between the expressions of PTEN and Survivin(r=-0.235, P＜0.05). Thesedata indicated that the expression of proteins is important to the carcinogenesis andprognosis of esophageal carcinoma.4. In the ESCC,carcinoma with adenoid structure and small cell carcinoma, theexpression of PTEN,MGMT,Survivin and IPO-38 was significantly different, andInvasion depth and the positive rate of the lymph node metastasis were bothsignificantly different, Which indicated that there are different carcinogenesis andbiological behavior in the ESCC,carcinoma with adenoid structure and small cellcarcinoma.
|
PDF Full Text Request