文献类型：期刊文献

中文题名：一种针对mdr1基因的寡核苷酸-阿霉素偶联物 :对KB-A-1细胞的毒性及其抑制P-gp蛋白表达的效果(英文)

英文题名：A novel antisense oligodeoxynucleotide-doxorubicin conjugate against the multidrug resistance gene mdr1 : inhibition to KB-A-1 cell and P -glycoprotein expression

作者：詹晓云[1];任宇红[1];张强[1];卢艳花[1];魏东芝[1];刘建文[1]

机构：[1]上海华东理工大学

年份：2004

卷号：9

期号：4

起止页码：403

中文期刊名：中国临床药理学与治疗学

外文期刊名：Chinese Journal of Clinical Pharmacology and Therapeutics

收录：CSTPCD;;CSCD:【CSCD2011_2012】；

基金：ProjectsupportedbytheNationalNaturalFoundation (№ 3 0 1710 88)andNa tionalPlan 863 (№ 2 0 0 1AA2 15 2 61)

语种：中文

中文关键词：反义寡核苷酸;偶联物;细胞毒性;多药抗性;P-gp蛋白

外文关键词：oligodeoxynucleotide; conjugate; cytotoxicity; multidrug resistance; P -glycoprotein

摘要：目的 :癌细胞膜上P gp糖蛋白的过量表达是肿瘤多药抗性的主要机制。人体内编码P gp糖蛋白的基因中仅有mdr1涉及多药抗性。本研究中设计了针对mdr1的反义核酸与阿霉素的偶联物 ,并且对其细胞毒性进行了考察。同时对偶联物对人表皮癌细胞株KB A 1内的P gp蛋白的表达也做了分子水平上的研究。方法 :使用MTT法考察偶联物对KB A 1细胞的毒性。用HPLC考察偶联物对细胞内阿霉素的积累量的影响。对于P gp蛋白表达的变化 ,主要是通过RT PCR及WesternBlot方法进行了研究。结果 :偶联物的细胞毒性比寡核苷酸高。在低剂量的偶联物 (0 .5 μmol·L-1)的作用下 ,细胞对阿霉素的敏感性提高。偶联物能有效的提高细胞内阿霉素的积累量。并且从RT PCR及WesternBlot看 ,偶联物处理后的细胞内P gp表达最少。结论 :选用合适的基团 ,对反义核酸进行结构修饰能够较好的增强反义核酸的性能。采用阿霉素作为偶联基团尽管增强了细胞毒性 ,但是在更大程度上增强了其抑制P gp蛋白的效力 ,提高了肿瘤耐药性的逆转倍数 ,具有一定的潜在应用价值。
AIM : Over-expression of P -glycoprotein ( P -gp) on the surface of tumor cells that works as a membrane pump enhancing the drug efflux is regarded as a main candidate mechanism of multidrug resistance (MDR). And mdr1 is one of the two main genes that encode P -gp in human cells. In this study, a novel conjugate made of mdr1 -anti-sense oligodeoxynucleotide (ODN) and a potent anticancer drug doxorubicin was constructed. The cytotoxicity of the conjugate as well as the effect of the conjugate on the modulation of P -gp-mediated MDR in KB-A-1 cell lines was studied. The molecular mechanism of its regulation effect was also investigated. METHODS : Multidrug resistant KB-A-1 cell lines were used. MTT was applied to measure the cytotoxicity of the conjugate and doxorubicin to KB-A-1 cells. Effect of the conjugate on intracellular doxorubicin accumulation was determined by HPLC. RT-PCR and Western blot were used to determine the expression of mdr1 gene and P -glycoprotein in KB-A-1 cells. RESULTS : The results showed that the conjugate exhibited more cytotoxicity than ODN, and also it could promote the cytotoxicity of doxorubicin to KB-A-1 cells. The IC 50 of doxorubicin dropped from 24 to 18 μg·ml -1. The results of RT-PCR and Western blot analysis illuminated that conjugate might down-regulate the P -gp expression. CONCLUSION : All the findings suggest that the conjugate is more potential for efficient reversal of MDR phenomenon than free ODN. The study indicated that such a conjugate should be a new feasible and efficient anti-sense reagent both in lab research and further in clinical therapy.