Reck And Mmp-14 In Children With Neuroblastoma And Failover Occurs In The Study On The Mechanism Of Action

PartⅠExpression of reversion-inducing-cysteine-rich protein with Kazal motifs,matrix metalloproteinase-14 and their correlations in neuroblastomaObjective:To detect the expression of RECK(reversion-inducing-cysteine-rich protein with Kazal motifs),MMP-14(matrix metalloproteinase-14),as well as their correlations in neuroblastoma and to investigate their roles in metastasis and development of the tumor.Methods:PV-6000 immunohistochemistry was used to measure the expression of RECK, MMP-14,in 36 samples ofneuroblastoma tissue selected from paraffin wax-embodied specimens with complete clinicopa-thological data,and another 18 cases of ganglioneuroma tissue.Results:The expression rate of RECK in neuroblastoma was low(16.7%),and reduced with the increased invasive depth and clinical stages,occurrence of lymphatic and long-distance metastasis(P＜0.05).The expression rate of MMP-14 in neuroblastoma was high (58.3%),and elevated with the increased invasive depth and clinical stages,and occurrence of lymphatic metastasis(P＜0.05).The expression of RECK was negatively correlated with the expression ofMMP-14(r=-0.418,P＜0.05).Conclusion:RECK is lowly expressed in neuroblastoma,but MMP-14 is highly expressed.RECK,MMP-14 may be served as the markers in the estimation of metastasis and dissemination of neuroblastoma. PartⅡThe establishment of the bear-tumor nude mice model and cell suspensionObjective:we use neuroblastoma patients'fresh tumor specimens to establish the human neuroblastoma cell lines and then establish different concentration of cell suspension to be inoculated in nude mice,to bulid the bear-tumor nude mice model,and further set up the mouse primary tumor cell line and the mouse metastatic tumor cell line.Method:Use the fresh tumor tissue to establish the in vitro cell line and use the cell line established by ourselves to get the bear-tumor nude mice model.Take the fresh tumor tissue from the patient under the sterilizing situation during the operation,used the enzyme digestion method to establish the in vitro cell line.Used the enzyme digestion method and erasion method to get the single cell suspension.Result:A total of ten nude mice has tumor,five mice have tumor at unilateral subcutaneous,two mice have tumor at bilateral retroperitoneal,three mice have systemic metastasis(both have obvious liver metastasis and cachexia signs),through primary culture to establish tumor cell lines in situ and metastatic tumor cell lines,cells stained darker,irregularly shaped,some is tear drop-shaped,a small number is triangular shape, Compositae Mission-shaped growth,low differentiation.Conclusion:We have successfully set up neuroblastoma tumor-bearing mouse model, and successfully set up the mouse primary tumor cell line and the mouse metastatic tumor cell line,confirming our own neuroblastoma cell lines have partial tumorigenicity and transfer characteristics,in order to lay the foundation for the transfer mechanism of further explore neuroblastoma and to create a good start for the next step to detect neuroblastoma metastasis-associated gene expression differences through RT-PCR method. PartⅢExpression and Significance of the Matrix Metalloproteinase Inhibitor RECK Gene in tumor-bearing nude mice cell linesObjective:We detect the expression of RECK and MMP-14mRNA in the Mouse primary tumor cell line and the Mouse metastatic tumor cell line to compare both the expression differences and their clinical significance.Methods:We detect the expression of RECK and MMP-14mRNA in the Mouse primary tumor cell line and the Mouse metastatic tumor cell line through RT-PCR method.Results:The mRNA level of the RECK gene in the Mouse primary tumor cell line was lower than that in the Mouse metastatic tumor cell line.The mRNA level of the MMP-14 in the Mouse primary tumor cell line was higher than that in the Mouse metastatic tumor cell line.Conclusion:These results indicate that RECK and MMP-14 might play important role in the pathogenesis of Neuroblastoma.RECK and MMP-14 could be considered as the marker for predicting the Neuroblastoma metastasis and development.