反义核酸对人肺腺癌细胞多药耐药逆转的实验研究  

Reversal of multidrug resistance in lung adenocarcinoma resistant cell line A549/R by mdr1 antisense oligodeoxynucleotides in vitro

作　　者：陈良安[1] 刘又宁[1] 

机构地区：[1]中国人民解放军总医院呼吸科,北京100853

出　　处：《中华医学杂志》2000年第3期219-221,共3页National Medical Journal of China

摘　　要：目的　探讨反义核酸对人肺腺癌细胞多药耐药的逆转效果。方法　以mdr1cDNA -9— +6为靶位点 ,合成反义核酸 (ATCCATCCCGACCTC)并用正义链作为对照 (GAGGTCGGGATGGAT) ,通过脂质体将其导入肺腺癌细胞A5 49/R ,分别采用逆转录聚合酶链反应、流式细胞仪、罗丹明试验及药敏试验观察两组细胞的mdr1mRNA、P糖蛋白 (Pgp)表达、罗丹明的聚集和对阿霉素的敏感性。结果　反义核酸处理的细胞mdr1mRNA下降 5 1 2 % ,正义组细胞和对照组细胞无明显变化、反义组细胞Pgp含量明显降低 ,细胞内罗丹明聚集 ,反义组 ,正义组和对照组的阿霉素IC50 分别为 0 0 0 9μg/ml、0 17μg/ml和 0 18μg/ml。 结论　反义核酸具有逆转人肺腺癌多药耐药的作用 ,其作用水平在mRNA或DNA水平 ,使Pgp的表达受到明显抑制 ,对阿霉素的敏感性明显提高。Objective To investigate the possibility of reversion of multidrug resistance (MDR) in lung adenocarcinoma resistant cell line A549/R by mdr1 antisense oligodeoxynucleotide. Methods A 15 mer phosphorothioate antisense oligodeoxynucleotide (ATCCATCCCGACCTC), complementary to -9～+6 in mdr1 cDNA sequence, was synthesized. Meanwhile, sense sequence (GAGGTCGGGATGGAT) was taken as control. Both antisense and sense oligodeoxynucleotides were transduced to A549/R cell line by lipofectin. The mdr1 mRNA, expression of Pgp, cellular rhodamine accumulation and sensitivity to daunorubicin were detected in antisense and sense oligodeoxynucleotides treated cells, using RT PCR, flow cytometry, rhodamine test and MTT method. Results Treatment with antisense oligodeoxynucleotide in A549/R cells led to a decrease in mdr1 mRNA and Pgp expression, an increase in rhodamine cellular accumulation, and a 20 fold increase of sensitivity to doxorubicin compared with those treated with sense oligodeoxynucleotide or control cells. Conclusion The mdr1 antisense oligodeoxynucleotide possesses an effect of reversing MDR in lung adenocarcinoma resistant cell A549/R by inhibiting mdr1 transcription and Pgp expression.

关 键 词：多药抗药性 肺肿瘤 反义核酸 

分 类 号：R734.2[医药卫生—肿瘤]