2017-11-012024-05-17https://scholars.lib.ntu.edu.tw/handle/123456789/694206摘要：醣蛋白P由MDR1/ABCB1基因所轉譯，經由ATP的驅動，是生物體排出多重藥物的轉運蛋白。MDR1/ABCB1基因的突變會影響到化療的成效或是其他醣蛋白P受質藥物的治療效果。本研究擬鎖定在犬隻MDR1基因的三個重要候選位點（單鹼基多樣性180, 單鹼基多樣性1664, 以及4鹼基缺失）的檢測。單鹼基多樣性180是位於MDR1基因的第一內含子(intron 1)靠近啟動子(promoter)的位置，若該位點的胸腺嘧啶(T)突變為鳥嘌呤(G)，MDR1基因會有過多的表現而導致生物體對藥物的抗藥性。而4鹼基缺失的突變會損害醣蛋白P的功能，當有醣蛋白P受質藥物的投予時，就會產生神經毒性。另外經由PolyPhen 和SIFT algorithms的程式預測，若單鹼基多樣性1664位點的鳥嘌呤(G)突變為腺嘌呤(A)，則會對醣蛋白P的功能有衝擊性的影響。過往對MDR1基因的檢測都是各自突變點的檢測，必須藉由引子作單一區段增幅之後，在作基因定序才能得知，曠日廢時；而未有全面整合性的研究。寡核酸微陣列是突變基因的新型高敏感檢測工具，而且具有同步多工的特性，可以同時檢測多種基因。藉由微陣列的探針和標的核酸的專一性雜交反應，使訊號的產出有如同基因定序的果效。尤有甚者，微陣列的檢測訊號可以直接由肉眼判讀，不需要影像偵測系統。本研究的目的是要發展一寡核酸微陣列的檢測平台，可快速同步地研究台灣地區犬隻MDR1基因的三個重要位點的突變概況。藉由統計分析了解臨床樣本基因型與化療藥物之間的關聯性，整合性地了解MDR1不同位置的基因型，對臨床藥物治療的影響。由於MDR1的基因型是化療藥物副作用以及多重藥物抗藥性的重要指標，因此在犬隻化療施予之前以及藥物投藥之前，皆強烈的建議必須作MDR1的基因型的檢測；而寡核酸微陣列的發展將是一個創新性高效能的整合性檢測平台。<br> Abstract: P-glycoprotein (p-gp), which is translated from MDR1/ABCB1 gene, is an ATP-driven multidrug efflux carrier. Mutations in the MDR1/ABCB1 gene can influence the effect of chemotherapy or the treatment of other p-gp substrate drugs. The detection of three canine candidate mutations in MDR1 gene, SNP180, SNP 1664 and 4-bp deletion, were chosen in this study. The SNP180 is located in the MDR1 intron 1 and near the promoter position. Mutation (T→G) could make MDR1 gene express more, leading to drug-resistance. The 4-bp deletion alters the expression or function of p-glycoprotein, resulting in neurotoxicosis after administration of the substrate drugs. And SNP1664 mutation (G→A) is predicted to have an impact on the p-glycoprotein function by PolyPhen and SIFT algorithms. The detection of MDR1 gene in the past targeted at each single mutation. The nucleotide segment was amplified using PCR and the mutation was confirmed via further sequencing. It is laborious and time consuming. No integrating and comprehensive survey has ever been done. Oligonucleotide microarray is highly sensitive to mutant gene detection and allows the analysis of multiple gene sequences to become possible. The specific complementary hybridization between probe and target nucleotides makes its effectiveness similar to sequencing. In addition, the hybridization results on microarrays could be clearly identified using the naked eye and no further imaging equipment is needed. The aim of this study is to develop oligonucleotide microarrays to efficiently detect the three significant mutations in MDR1 genes in dogs in Taiwan. A large number of clinical samples will be screened to verify the correlation between MDR1 genotypes and the side effect of chemotherapy medication using statistical methods. Because MDR1 genotypes would be useful prognostic indicators for the occurrence of adverse drug reactions and/or the development of multidrug resistance, the detection of MDR1 genotypes is strongly recommended before chemotherapy and medication initiation for dogs. The oligonucleotide microarray would be a novel and highly competent tool for simultaneous detection of multiple canine MDR1 genotypes.醣蛋白P犬MDR1基因寡核酸微陣列基因型caninegenotypeMDR1oligonucleotide microarrayp-glycoprotein