Sensitization and apoptosis augmentation of K562/ADM cells by anti-multidrug resistance gene peptide nucleic acid and antisense oligodeoxyribonucleotide  

抗多药耐药基因肽核酸与反义寡脱氧核苷酸增强K562/ADM细胞的敏感性和促进凋亡(英文)

作　　者：魏虎来 吴勇杰 景涛[2] 白德成[2] 马兰芳[3] 

机构地区：[1]甘肃省中药新药临床前研究重点实验室 [2]兰州医学院医学实验中心 [3]兰州医学院血液病研究所

出　　处：《Acta Pharmacologica Sinica》2003年第8期805-811,共7页中国药理学报（英文版）

基　　金：Project supported by Gansu Scientific and Technological Foundation for the Middle-Age and Youth (№ YS981-A23-010).

摘　　要：AIM: To investigate the reversal effect and apoptosis enhancement of peptide nucleic acid (PNA) and antisense oligodeoxyribonucleotide (ASODN) targeted to multidrug resistance gene (mdr1) on human multidrug resistant leukemia K562/ADM cells. METHODS: A 15-mer PNA and the same sequence of ASODN, complementary to the 5' end of the AUG initiator codon-containing region of mdr1 messenger RNA (MDR1-PNA, MDR1-ASODN), were designed and synthesized. Proliferation and sensitivity to adriamycin of K562/ADM cells treated with MDR1-PNA-and MDR1-ASODN were analyzed with a MTT colorimetric assay. Apoptotic morphologies, P-glycoprotein (P-gp) expression, intracellular adriamycin accumulation, and cell cycle were measured. RESULTS: MDR1-PNA 1 to 10 μmol/L and MDR1-ASODN 2 to 20 μmol/L alone had no inhibitory effects on the proliferation of K562/ADM cells, but significantly inhibited the growth of K562/ADM cells cultured in adriamycin-containing medium. After treatment with MDR1-PNA and MDR1-ASODN, intracellular adriamycin accumulation in K562/ADM cells increased greatly and P-gp synthesis was strikingly reduced. The resistance to adriamycin of the drug-resistant cells was partly reversed and the cells were induced to apoptosis by adriamycin. The reversal efficacy of MDR1-PNA was 3.1-fold higher than that of the same sequence of MDR-ASODN, but neither MDR1-PNA nor MDR1-ASODN could completely block the mdr1/P-gp expression. CONCLUSION: Sequence-special PNA targeted to mdr1 gene more effectively than the same sequence of MDR 1-ASODN inhibited the expression of P-glycoprotein to overcome the drug-resistance.目的:研究多药耐药基因mdr1特异性肽核酸(MDR1-PNA)和反义寡脱氧核苷酸(MDR1-ASODN)对耐药白血病K562/ADM细胞耐药性和凋亡抑制的逆转效应。方法:合成互补于mdr1基因mRNA的PNA和硫代ASODN,直接转染细胞,MTT比色法检测细胞增殖活性;流式细胞仪检测细胞周期和P-糖蛋白(P-gp)表达;激光共聚焦显微镜观察细胞内多柔比星(ADM)含量和分布。结果:MDR1-PNA 1-10 μmol/L和MDR1-ASODN2-20 μmol/L不抑制K562/ADM细胞的增殖,但下调K562/ADM细胞P-gp的表达,增加细胞内ADM的含量,提高K562/ADM细胞对ADM诱导凋亡的敏感性而逆转其耐药性。PNA的作用较ASODN高3.1倍,无论是PNA还是ASODN均不能完全 阻断P-gp的合成。结论:mdr1基因特异性PNA和反义ODN抑制耐药白血病细胞P-gp的表达而逆转其耐药性和凋亡抑制,PNA的活性高于相同序列的ASODN。

关 键 词：peptide nucleic acids antisense oligodeoxyribonucleotides MDR genes P-GLYCOPROTEIN multiple drug resistance APOPTOSIS LEUKEMIA DOXORUBICIN 

分 类 号：R733.7[医药卫生—肿瘤]