The Study Of Select Tumor Markers For Diagnosis And Treatment Of Solid Malignant Tumors In Childhood

PART 1:EXPRESSION AND SIGNIFICANCE OF MDR1,MRP,LRP IN PERIPHERAL BLOOD OF CHEMOTHERAPY CHILDREN WITH MALIGNANT SOLID TUMORS DETECTION BY FQ-PCRObjective: To investigate the expression of multi-drug resistance gene (MDR1),multi-drug resistance-associated protein gene (MRP),lung resistance protein gene (LRP) in peripheral blood mononuclear cells of chemotherapy children with malignant solid tumors and its relationship with clinical chemotherapy.Methods : Pathologically confirmed 59 cases of children with cancer ,Including neuroblastoma 24 cases, 16 cases of yolk sac tumor, lymphoma, 19 cases; 30 healthy children as control group.The expression of MDR1,MRP,LRP in peripheral blood mononuclear cells(PBMCs) before and after chemotherapy, fresh frozen tumor tissue and the normal control group peripheral blood were detected by real-time fluorescence quantitative PCR(FQ-PCR). Detection of tumor markers before and after chemotherapy in children with chemotherapy in conventional. The use of statistical methods to chemotherapy patients and the control group before chemotherapy peripheral blood MDR 1, MRP, LRP expression of mRNA for T test; Peripheral blood before and after chemotherapy in children with chemotherapy, MDR 1, MRP, LRP mRNA expression in resected tumor tissue and MDR1, MRP, LRP expression of the corresponding tumor marker level changes before and after chemotherapy, the correlation analysis.Results:Neuroblastoma group, yolk sac tumor before chemotherapy, significantly higher expression of MDR1, the difference was statistically significant (P <0.01); Lymphoma before chemotherapy, the detection rate of MDR1 and the control group showed no significant difference (P> 0.05); All chemotherapy patients in peripheral blood of MRP, LRP expression in the control group before chemotherapy was no significant difference (P> 0.05).All MDR1 expression levels in peripheral blood of chemotherapy children with the same period tumor tissue expression levels of MDR1 positive correlation ( r = 0.894, P = 0.000); There was an negative correlation between MDR1 mRNA level in peripheral blood before chemotherapy and the changes size of corresponding tumor markers after the first time chemotherapy, neuroblastoma of the urine VMA (r = -0.440, P = 0.046), yolk sac tumor of the AFP (r = -0.831, P = 0.000), lymphoma of the LDH (r = -0.495, P = 0.031). We found that increased MDR1 mRNA level in peripheral blood of chemotherapy children was related to increased the number of chemotherapy treatments(P<0.05).Conclusion:Detect the mdr1 level in PBMCs before and after chemotherapy is valuable for evaluating the treatment response. PART 2:THE STUDY OF NEUROBLASTOMA-RELATED TUMOR MARKERS DETECTObjective: Explore the use of real-time fluorescence quantitative PCR (FQ-PCR) quantitative detection of BCL-2 gene mRNA in neuroblastoma (NB) expression and use ELISA to detect neuroblastoma peripheral blood VEGF, NSE, SF expression, and with neuroblastoma, development and prognosis.Methods:Children with neuroblastoma collected surgical specimens from 16 cases, -80℃preserved specimens of which 6 cases before chemotherapy, after chemotherapy specimens in 8 cases, both specimens before chemotherapy and another 2 patients after chemotherapy, and set the time acquisition peripheral blood of children, collected by centrifugation plasma -80℃save. Application of real-time fluorescence quantitative PCR detection of BCL-2 for all samples and N-MYC gene mRNA, while the corresponding tumor specimens collected paraffin sections, for immunohistochemistry. Check the use of ELISA method in children with neuroblastoma in peripheral VEGF, NSE, SF expression. Evans criteria used neuroblastoma in children with clinical stage, all tissue samples were carried out Shimade type. Comparison of the use of statistical methodsⅠ~ⅡandⅣS phase and stageⅢ~Ⅳ, F-type and UF type chemotherapy before and after chemotherapy in neuroblastoma tissues of BCL-2 gene expression levels of mRNA size and expression of BCL-2 protein The size of the positive rate; Analysis the correlation of BCL-2 expression and N-MYC gene and resistance gene expression Correlation; before and after chemotherapy, surgery of peripheral blood VEGF, NSE, SF of expression; BCL-2, VEGF, NSE, SF and neuroblastoma occurrence, development and prognosis.Results:Real-time fluorescence quantitative PCR detection of BCL-2 gene mRNA in neuroblastoma tissue volume in stageⅢ~Ⅳwas significantly higher thanⅠ~ⅡandⅣS (P <0.05), UF-type was significantly higher than F-type (P < 0.05), the expression of after higher than before chemotherapy (P <0.05), results of immunohistochemistry and PCR results are in agreement; BCL-2 expression and the copy number of N-MYC was significantly correlated (P <0.05), and the resistance gene's expression was not significantly related (P> 0.05); BCL-2, VEGF, NSE, SF and the expression of neuroblastoma in children was related to prognosis (P <0.05).Conclusion:BCL-2 as a key apoptosis genes, using real time quantitative PCR detection of neuroblastoma in the expression, could be used as biological characteristics of neuroblastoma cells and an important indicator of prognosis. VEGF, NSE, SF as a tumor marker in peripheral blood can be used as chemotherapy monitoring and an important indicator of prognosis.