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Preparation, Identification and Preliminary Clinical Application of Polyclonal Antibody of Atra-related Resistance Gene HA117

Author: ChenJianFei
Tutor: JinXianQing
School: Chongqing Medical University
Course: Pediatrics
Keywords: HA117 gene Polyclonal antibody Preparation Appraisal HA117 MDR1 Tumor resistance Staging Lymph node metastasis
CLC: R739.4
Type: Master's thesis
Year: 2011
Downloads: 10
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Abstract

Purpose: Preparation and characterization of HA117 基因多克隆 antibodies. Methods: pAdTrack-HA117 HA117 expression plasmid as a template, PCR amplification HA117 gene, PCR products were characterized by restriction enzyme digestion, and PET-28a recombinant plasmid PET-HA117 created and transferred to the BL21 (DE3) competent cells for protein HA117 induced expression, SDS-PAGE using the expression product detected. New Zealand white rabbits were immunized with purified, polyclonal antibodies. Through its indirect ELISA titer. Transfected HEK293 cells recombinant adenovirus carrying HA117 for the experimental group, untransfected HEK293 cells as the control group, with immunofluorescence and Western blot was used to detect protein overexpression HA117, HA117 基因多克隆 verify antibodies prepared specificity. Result: successful HA117 基因多克隆 antibodies by indirect ELISA assay showed that anti-HA117 serum titer of 1:729 000. Immunofluorescence and Western blot showing prepared HA117 HA117 polyclonal antibody can specifically recognize the antigen protein. Conclusion: The higher the potency and specificity of HA117 polyclonal antibodies can be used for immunohistochemistry, Western blot method to clinical testing HA117 protein expression. Objective: To study HA117 protein in neuroblastoma expression and its clinical significance. Methods: 20 cases of neuroblastoma as the research object, take the tumor tissue 14 cases as controls, using immunohistochemical SP method to observe its expression. Method according to Shimada staging neuroblastoma into the FH-type (Favorable Histology Group) and uFH type (Unfavorable Histology Group) are two HA117 were detected in the expression differences. According to lymph node metastasis and distant metastasis divided into groups with and without metastasis, analysis HA117 protein expression differences. Detected 20 cases of neuroblastoma HA117 expression of MDR1 expression and differences and to analyze its clinical significance. Results: HA117 in neuroblastoma expression was 70% (14/20), the expression in tumor tissue was 21% (3/14), and the results were statistically significant. FH 10 cases, there are five cases HA117 positive expression, the positive rate was 50%, uFH type in 6 cases, 6 cases of all, the positive rate was 100%, uFH positive rate in FH type. With lymph node and distant metastasis in 11 cases, HA117 expression in 11 cases, the positive rate of 100%, 9 patients without metastasis, there are three cases of expression, expression was 33%. 20 cases of neuroblastoma in HA117 expression rate of 70%, the result was statistically significant. 20 cases of neuroblastoma in, MDR1 expression rate of 60%, HA117 positive rate was 70%, slightly lower than HA117 expression positive rate, but P gt; 0.05, the result was not statistically significant. Conclusion: 1.HA117 proteins in neuroblastoma has a broad distribution, all patients were preoperatively using ATRA-induced differentiation, suggesting that neuroblastoma multidrug resistance phenomenon. 2 cases with lymph node and distant metastasis HA117 was significantly higher than that without lymph node metastasis. 3 pathological stage and tumor histology was no significant difference.

Preparation, Identification and Preliminary Clinical Application of Polyclonal Antibody of Atra-related Resistance Gene HA117

Abstract

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