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The Anti-human ADAM15Antibody Exhibited Anti-proliferation and Tumor Targeting Effects in Cancer

Author: LiYaHui
Tutor: YangBingHua;HeYang;ZhangBin
School: Suzhou University
Course: Clinical Laboratory Science
Keywords: tumor cell proliferation ADAM15 Antibody Iodine-125 Radioimmunoimaging
CLC: R73-36
Type: Master's thesis
Year: 2012
Downloads: 19
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Abstract


Purpose: A disintegrin and metalloproteinase15(ADAM15)is one kind oftransmembrane glycoprotein recently discovered which has a close relationship withcancer.This study investigated the biological characteristics of anti-ADAM15antibodyabout its combination with different cancer cells as well as its anti-proliferation effect andalso intended to initially study its ability of tumor targeting in vivo in order to find a novelmethod to treat cancer.Methods:(1)Anti-ADAM15antibody was purified using an immune affinitychromatography method;(2) The combinations of this antibody with lung cancer cellA549,gastric cancer cell HGC-27,and vascular endothelial cell HMEC-1were detectedunder different time(2,24,48h) or different concentrations(0,2,4,8μg/ml) using an indirectELISA method.(3)Cell proliferation inhibition experiment was administrated using anMTT method in order to investigate the effect of this antibody on cancer cell growth whenthe concentration was4μg/ml and reaction time was48,72,96h.(4) The anti-humanADAM15polyclonal antibody was labeled with I-125using Chloramine-T method andinjected into a nude mice bearing gastric carcinoma xenografts in order to conduct theradioimmunoimaging assay to evaluate its tumor targeting ability in vivo.Results:1.When the final concentrations of this antibody were from2to8μg/ml, anti-ADAM15antibody effectively combined with both tumor cell and vascular endothelial cell;the combination became stronger when the time of reaction was long enough (24or48h)or the concentration of this antibody became higher.2.①When the final concentration was4μg/ml,anti-ADAM15antibody exhibited anti-proliferation effects in all the three cell lines tested:all the three cell lines showed alower growth rate compared with the control groups when the culture time was48or72h;when the culture time was96h long,only HMEC-1cell growth rate was lower than itscontrol group.②The inhibition ratios of HGC-27and HMEC-1groups were higher thanthe A549group when the culture time was48h, with inhibiton ratios (25.74±5.93)%,(23.32±3.70)%,(13.68±4.33)%respectively, the difference had significance ofstatistics(P<0.01).③When the culture time was72h long, the inhibition ratios of HGC-27and HMEC-1cell lines became lower compared with the48h results,with inhibitionratios (14.11±6.25)%,(17.81±4.11)%respectively, the difference had significance ofstatistics(HGC-27group P<0.01,HMEC-1group P<0.05).3.The labeling efficiency of125I-anti-ADAM15antibody was (75.16±9.43)%and itsradiochemical purity was (99.44±0.21)%. Tumors could be cleanly visualized in SPECTplanar images, and the radioactivity ratio of tumor to non-tumor tissue was3.84±0.43at48h post-injection.Conclusion:Anti-ADAM15antibody can effectively combine with gastric cancer cell,lung cancercell and vascular endothelial cell,moreover,this antibody can inhibit the proliferation ofthese three cell lines.125I-anti-ADAM15antibody can target the gastric carcinoma in vivo,and provide good radioimmunoimages.

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