The Study On The Correleation Between Expressions Of PTEN,VEGF,CD105 And The Occurence, Invasion And Metastasis Of Esophageal Squamous Cell Carcinoma

[Background and objective]PTEN was descovered in 1997 .Now it is identified as a new tumor suppressor gene and plays an important role in regulation of cellular proliferation, differentiation and apoptosis, and it may participate the carcinogenesis , development and the formation of vessels of tumor. Now PTEN has become a focus in medical research.Esophageal carcinoma is a kind of common diseases in china especially in Henan province. It was the fourth in the death rate of malignant tumor in china. Most cases causing the death of esophageal carcinoma are tumor's invasion and metastasis. So exploration about invasion and metastasis of esophageal carcinoma has become a focus.Recently, It has been found that losing of PTEN gene activity has a relationship with the occurrence ,development of many tumor. Furthermore, some researchers have proved that PTEN can suppress the growth, invasion and metastasis of tumor cells by suppressing the formation of vessels. Up to the present, there are very few studies of expression of PTEN mRNA in esophageal squamous cell carcinoma,thestudy of the expressions of PTEN> VEGF, CD105 in esophageal squamous cell carcinoma is even more few. There is no report about correlation between the expression of PTEN > VEGF> CD 105 and the carcinogenesis, invasion and metastasis of esophageal squamous cell carcinoma. In order to further explore the relation between the expression of PTEN and the carcinogenesis, invasion and metastasis of esophageal squamous cell carcinoma , to find early defective markers of invasion and metastasis of esophageal carcinoma and to find out useful ways to suppress the invasion and metastasis of esophageal carcinoma, this article detects the expression of PTEN protein and mRNA in 49 cases of esophageal squamous cell carcinoma and normal mucosa by immunohistochemistry and RT-PCR way. This article intends to detects the expression of PTEN protein in 49 cases of tumor-adjacent mucosa tissues by immunohistochemistry, and the expression of vascular endothelial growth factor (VEGF) in 49 cases of tumor-adjacent mucosa tissues by immunohistochemistry and measures the micro vessel density (MVD) in 49 cases of esophageal squamous cell carcinoma by anti-CD 105 antibody, and explores the correlation between PTEN and VEGF> CD 105. Through the above tests, the article tries to describe the significance of the expression of PTEN on carcinogenesis, development, invasion and metastasis of esophageal squamous cell carcinoma. Furthermore, it also tries to explore whether the mechanism of PTEN may suppress the carcinogenesis, development, invasion and metastasis of esophageal squamous cell carcinoma has some relation with suppression of the expression of VEGF, decreasing the MVD in tumor tissue. These findings can set a theoretical foundation for exploring the mechanism of carcinogenesis, invasion, metastasis of esophageal squamous cell carcinoma and finding out useful ways to suppress occurence, development of esophageal squamous cell carcinoma. [Materials and methods]49 cases of esophageal squamous cell carcinoma were collected in the First Affiliated Hospital, Zhengzhou University and the Second Affiliated Hospital, Zhengzhou University. All cases has no the history of the chemotherapy .radiotherapy and immunotherapy. All cases were identified by pathologist, it is entirely esophageal squamous cell carcinoma.20cases are I rank, 18 cases are II rank, 11 cases are IIIrank. 18 example appeared the lymph node metastasis,31 cases are non- lymph node metastasis. 16 cases are on the low layer invaded and 33 cases on the deep layer invaded. Each all in site of tumor and far-tumor normal mucoma cut a piece of tissue. The expression of PTEN protein and mRNA were detected in 49 cases of esophageal squamous cell carcinoma and normal mucosa by immunohistochemistry and RT-PCR way. The expression of PTEN protein were detected in 49 cases of tumor-adjacent mucosa tissues by immunohistochemistry. Meanwhile, using immunohistochemistry detected the expression of vascular endothelial growth factor (VEGF) in 49 cases of tumor-adjacent mucosa tissues and the micro vessel density (MVD) in 49 cases of esophageal squamous cell carcinoma by anti-CD 105 antibody. All the data were analyzed by SPSS 11.0 stastistical package, the count information calculated the positive rate and enumeration data were expressed by theMean±standard deviationC x±s).The comparision of positive rate uses the Chi-square,the mean of two groups uses t-test, the mean of more than two groups uses ANVOA. The relation of two variable are analyzed by correlation analysis. The level of significant difference was a=0.05. [Results]1. The positive rates of expression of PTEN mRNA in esophageal squamous cell carcinoma and normal esophageal tissues were 44.9% and 81.6% respectively, a significant difference between them (PO.05); With the increase of invasion depth, the positive rates of expression of PTEN mRNA in esophageal squamous cell carcinoma decreased clearly (PO.05); the positive rates of expression of PTEN mRNA in the group with lymph node metastasis was 22.2%. While in the group without lymph node metastasis, it was 58.1%, a significant difference between them (PO.05) . the positive rates of expression of PTEN mRNA in esophageal squamous cell carcinoma of grade K II and III were 61.1 %> 38^/ck and 36.4% respectively, no significant difference among them (P>0.05) .2. The positive counts(the brightness value of band) of the expression of PTEN mRNA in the tissues of esophageal squamous cell carcinoma and normal esophagealmucosa were 0.56±0.23 and 0.81±0.36 respectively, a significant difference between them (PO.05) . The positive counts of the expression ofPTENmRNA of grade I, II and III of the esophageal squamous cell carcinoma were 0.61±0.18, 0.49±0.35 and 0.41±0.61 respectively, no significant difference among them (P>0.05) . In the tissues of esophageal squamous cell carcinoma, the positive counts of the expression of PTEN mRNA on the low layer invaded and the deep layer invaded were 0.66±0.18 and 0.34±0.18, a significant difference between them. The positive counts of the expression of PTEN mRNA in lymph node metastasis group and nonlymph node metastasis group were 0.43±0.60 and 0.57±0.23 respectively, no ignificant difference between them (P>0.05) .3. The positive rates of expression of PTEN protein in esophageal squamous cell carcinoma and normal esophageal tissues were 53.1% and 100.0% respectively, a significant difference between them( .PO.05); With the increase of invasion depth and malignancy, the positive rates of expression of PTEN protein in esophageal squamous cell carcinoma decreased clearly (P<0.05); the positive rates of expression of PTEN protein in the group with lymph node metastasis was33.3%. While in the group without lymph node metastasis, it was 64.5%, a significant difference between them(PO.05) .4.The positive rates of expression of PTEN protein in tumor-adjacent normal epithelium and simple hyperplasia epithelium, mild atypical hyperplasia epithelium, severe atypical hyperplasia epithelium, carcinoma in situ and invasive carcinoma were 100.0%, 81.0%, 69.2%> 66.7%, 66.7% and 53.1% respectively, a significant difference between the simple hyperplasia epithelium, mild atypical hyperplasia epithelium, severe atypical hyperplasia epithelium, carcinoma in situ, invasive carcinoma and the normal epithelium.5. The rates of the expression of VEGF protein in normal epithelium and simple hyperplasia, mild dysplasia, severe dysplasia, carcinoma in situ of the esophageal mucosa of tumor-adjacent and invasive carcinoma were 0.0%, 0.0%, 15.4%, 44.4%, 66.7% and 62.3% respectively, and there was a significant difference between thesevere dysplasia, carcinoma in situ, invasive carcinoma and the normal epithelium.6. The positive rates of the expression of VEGF protein in the lymph node metastasis group and nonlymph node metastasis group were 83.3% and 54.4% respectively, a significant difference between them (PO.05) .while the positive rates of the expression of VEGF protein in grade I, II and III of esophageal squamous cell carcinoma were 55.0% > 66.7% and 81.8%,respectively, no significant difference among them (P>0.05) . In the tissues of esophageal squamous cell carcinoma, the positive rates of the expression of VEGF protein on low layer invaded and deep layer invaded were 69.7% and 56.3%,respectively,no significant difference between them., too (P>0.05) .7. The means of MVD marked with CD 105 antibody in the lymph node metastasis group and nonlymph node metastastic group were 45.27±6.54 and 31.57±9.39 respectively,also a significant difference between them (PO.05 ). The means of MVD on the low layer invaded and the deep layer invaded tissues of esophageal squamous cell carcinoma were 34.47±11.69 and 40.00±8.84, a significant difference between them (PO.05) . The means of MVD in grade 1,11 and Illof esophageal squamous cell carcinoma were 36.92±10.85 > 37.65±9.50 and 38.06±12.19,respectively, significant difference among them (P>0.05)8. In esophageal squamous cell carcinoma, the positive rate of the expression of PTEN mRNA was lower than that of the expression of PTEN protein .There was a significant difference between them (PO.05) , and there was a positive correlation between them (PO.05) . ■9. There was a negative correlation between the positive expression rate of PTEN protein and the expression of VEGF protein in esophageal squamous cell carcinoma. The positive expression group of PTEN protein and negative group in esophageal squamous cell carcinoma, the positive expression rate of VEGF were 50.0% (13/26) and 82.6% (19/23 ) respectively, a significant difference between them (PO.05) .10. The positive expression group of PTEN protein and negative group in esophageal squamous cell carcinoma, the means of MVD were 35.76±9.42 and 41.59±10.80, asignificant difference between them (PO.Ol) .11. The positive expression group of VEGF protein and negative group in esophagealsquamous cell carcinoma, the means of MVD were 41.99±9.83 and 31.07±6.38, asignificant difference between them (.PO.01) .[Conclusion]1 .The positive expression of PTEN protein and mRNA in esophageal squamous cellcarcinoma were both lower than that of PTEN protein and mRNA in normalesophagus mucosa, and the positive expression of PTEN protein in tumor-adjacentatypical hyperplasia epithelium had a trend of decreasing gradually. It hints that lossor decreased expression of PTEN maybe play an important role in the tumorigenesisand development of esophageal squamous cell carcinoma.2.It indicated that PTEN gene might contribute to invasion and metastasis ofesophageal squamous cell carcinoma and its loss or decreased expression had closelycorrelation with invasion depth and lymph node metastasis of esophageal squamouscell carcinoma. The positive counts of the expression of PTEN mRNA in lymph nodemetastasis group was lower than that in nonlymph node metastasis group ,but nosignificant difference between them.3. There was a correlation between the expression of PTEN protein and pathologicalgrade of tumor. It implied that PTEN gene made great contribution to cellulardifferentiation.4.There was a positive correlation between the expression of PTEN protein and theexpression of PTEN mRNA. It implied that the expressions of PTEN mRNA andprotein had consistency. The expression of PTEN mRNA was lower than theexpression of PTEN protein, it showed there were some regulations of PTEN proteinlevel, or heighten of stability of PTEN protein.5.hi tumor-adjacent mucous membrane, the rates of the expression of VEGF proteinincreased gradually from simple hyperplasia to severe atypical hyperplasiaepithelium. It implied that the expression of VEGF protein correlated with thetumorigenesis of esophageal squamous cell carcinoma.6.The expression of VEGF protein was no associated with pathological grade andinvasion depth. But it was associated with lymph node metastasis of esophageal squamous cell carcinoma, it implied that VEGF protein can reflect potential of lymph node metastasis of esophageal squamous cell carcinoma.7.1n esophageal squamous cell carcinoma, the means of MVD was no associated with pathological grade, but it was associated with lymph node metastasis and invasion depth of esophageal squamous cell carcinoma. It implied that MVD was a target to esophageal squamous cell carcinoma,can be used to estimate curative effect and prognosis.8.There was a negative correlation between the expression of PTEN protein and the expression of VEGF protein, the means of MVD. There was a positive correlation between the expression of VEGF protein and the means of MVD. It implied that the loss or decreased expression of PTEN protein can promote the formation of vessels of tumor by ascending the expression of VEGF protein, accordingly, offering the precondition for growth and metastasis of esophageal squamous cell carcinoma. United examining PTEN,VEGF and MVD can become a objective target to evaluate potential of lymph node metastasis of esophageal squamous cell carcinoma, it had a great significance to the judgement of prognosis in esophageal squamous cell carcinoma.