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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。
AIM: To investigate the reversal effects of MDR1-PNA and MDR1-ASODN on the drug resistance and apoptosis of drug-resistant leukemia K562 / ADM cells. METHODS: PNA and thio ASODN, which were complementary to mRNA of mdr1 gene, were synthesized and directly transfected into cells. MTT assay was used to detect cell proliferation. Flow cytometry was used to detect cell cycle and P-glycoprotein (P-gp) expression. The content and distribution of doxorubicin (ADM) in the cells were observed under a confocal microscope. Results: MDR1-PNA 1-10 μmol / L and MDR1-ASODN2-20 μmol / L did not inhibit the proliferation of K562 / ADM cells but down-regulated the expression of P-gp, increased the intracellular ADM content and increased the expression of K562 / ADM cells to ADM induced apoptosis and reverse its resistance. The effect of PNA is 3.1 times higher than that of ASODN, and neither PNA nor ASODN can completely block the synthesis of P-gp. CONCLUSION: The mdr1 gene-specific PNA and antisense ODN inhibit the expression of P-gp in drug-resistant leukemia cells and reverse the drug resistance and apoptosis inhibition. The activity of PNA is higher than that of ASODN with the same sequence.