用针对mdr1信使核糖核酸的反义寡脱氧核苷酸调节多药耐药性。

Modulation of multidrug resistance with antisense oligodeoxynucleotide to mdr1 mRNA.

作者信息

Sola J E, Colombani P M

机构信息

Department of Surgery, Johns Hopkins University, Baltimore, Maryland, USA.

出版信息

Ann Surg Oncol. 1996 Jan;3(1):80-5. doi: 10.1007/BF02409056.

DOI:10.1007/BF02409056

PMID:8770307

Abstract

BACKGROUND

P-glycoprotein (pgp), a 170-kDa adenosine triphosphate-dependent membrane drug efflux pump encoded by the mdr1 gene, mediates cross-resistance in tumor cells to structurally unrelated cancer drugs. We investigated the capacity for modulating multidrug resistance by selectively inhibiting synthesis of Pgp using an antisense oligodeoxynucleotide complementary to the initiation codon of mdr1 messenger RNA.

METHODS

By continuous culture of K562 in 100 nM vincristine, a resistant cell line, K562/VCR100, was derived with high expression of Pgp (95.9% of cells) and an IC50 40-fold greater than that of the parental cell line. The K562/VCR100 cells were treated with 10 microM of 15-mer antisense and sense phosphorothioate oligodeoxynucleotides. Modulation of multidrug resistance was analyzed using a daunorubicin/tritiated thymidine incorporation assay and flow cytometric assessment of cellular rhodamine 123 accumulation.

RESULTS

Treatment of K562/VCR100 with the antisense oligodeoxynucleotide led to a doubling in daunorubicin growth inhibition at 1 microgram/ml and a tripling of growth inhibition at 0.6 micrograms/ml (p < 0.0023); a 58% reduction in the daunorubicin IC50 (p < 0.02); and an increased rate of rhodamine-123 accumulation (p = 0.02) compared with treatment with sense oligodeoxynucleotide or media controls.

CONCLUSIONS

These results suggest that antisense oligodeoxynucleotides may serve as a useful adjunct in the treatment and prevention of multidrug resistance during cancer chemotherapy.

摘要

背景

P-糖蛋白（pgp）是一种由mdr1基因编码的170 kDa三磷酸腺苷依赖性膜药物外排泵，介导肿瘤细胞对结构不相关的抗癌药物产生交叉耐药性。我们研究了使用与mdr1信使核糖核酸起始密码子互补的反义寡脱氧核苷酸选择性抑制Pgp合成来调节多药耐药性的能力。

方法

通过在100 nM长春新碱中连续培养K562，获得了一个耐药细胞系K562/VCR100，其Pgp高表达（95.9%的细胞），IC50比亲代细胞系大40倍。用10 microM的15聚体反义及正义硫代磷酸寡脱氧核苷酸处理K562/VCR100细胞。使用柔红霉素/氚标记胸腺嘧啶掺入试验和细胞罗丹明123积累的流式细胞术评估来分析多药耐药性的调节。