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目的采用一段人工合成的互补于人ｍｄｒ１基因的反义硫代硫酸脱氧寡核苷酸（简称反义核酸）转染白血病多药耐药细胞株Ｋ５６２／ＡＤＭ，来逆转白血病细胞的多药耐药。方法ＭＴＴ法检测Ｋ５６２／ＡＤＭ细胞对阿霉素的ＩＣ５０，流式细胞仪分析细胞内的柔红霉素含量，免疫细胞化学染色方法确定Ｐ１７０的表达水平，ＲＴ－ＰＣＲ检测ｍｄｒ１－ｍＲＮＡ的相对水平（ｍｄｒ１／β２－ＭＧ）。结果浓度为２μｍｏｌ的反义核酸使Ｋ５６２／ＡＤＭ细胞对阿霉素的ＩＣ５０从处理前的２５．７２μｇ／ｍｌ降至处理后的６．９７μｇ／ｍｌ，相对逆转效率为７３．０１％。反义核酸亦使Ｋ５６２／ＡＤＭ细胞内的柔红霉素含量增加，Ｐ１７０表达阳性率从处理前的９８．６％±１．０％降至处理后的１８．３％±０．７％，ｍｄｒ１／β２－ＭＧ下降了０．５，ｍｄｒ１－ｍＲＮＡ相对水平有所下降。结论反义核酸通过下调ｍｄｒ１基因，阻断Ｐ１７０的表达，从而降低Ｋ５６２／ＡＤＭ的耐药性。
Objective To transfect leukemia multidrug resistant cell line K562/ADM by artificially synthesizing antisense thiosulfate deoxynucleotide complementary to human mdr1 gene to reverse the multidrug resistance of leukemia cells. . Methods The IC50 of K562/ADM cells against adriamycin was detected by MTT assay. The content of daunorubicin in cells was analyzed by flow cytometry. The expression level of P170 was determined by immunocytochemical staining. The relative level of mdr1-mRNA was detected by RT-PCR. (mdr1/β2-MG). Results The concentration of 2μmol of antisense nucleic acid reduced the IC50 of K562/ADM cells against adriamycin from 25.72μg/ml before treatment to 6.97μg/ml after treatment, and the relative reversal efficiency was 73.01%. Antisense nucleic acids also increased the content of daunorubicin in K562/ADM cells, and the positive rate of P170 expression decreased from 98.6%±1.0% before treatment to 18.3%±0.7% after treatment. Mdr1/β2-MG decreased by 0.5, and the relative level of mdr1-mRNA decreased. Conclusion The antisense nucleic acid can down-regulate the mdr1 gene and block the expression of P170, thus reducing the drug resistance of K562/ADM.