生存素表达的下调逆转了阿霉素耐药HL-60/ADR细胞系中的多药耐药性。

Down-regulation of survivin expression reversed multidrug resistance in adriamycin-resistant HL-60/ADR cell line.

作者信息

Wang Lei, Zhang Gui-Mei, Feng Zuo-Hua

机构信息

Department of Biochemistry and Molecular Biology, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.

出版信息

Acta Pharmacol Sin. 2003 Dec;24(12):1235-40.

PMID:14653950

Abstract

AIM

To investigate the ability of an antisense RNA eukaryotic expression plasmid pcDNA3.1/survivin in down-regulating the expression level of survivin mRNA and survivin protein and reversed multidrug resistance (MDR) in adriamycin-resistant HL-60/ADR cell line.

METHODS

The expression of survivin mRNA was measured by RT-PCR and the expression of survivin protein was measured by Western blot. Caspase-3 activity was determined by Phar Mingen colorimetric assay kit. Apoptosis was assessed by flow cytometry. The chemosensitivity of HL-60/ADR cells to adriamycin (ADR) was measured by MTT assay.

RESULTS

pcDNA3.1/survivin down-regulated the expression level of survivin mRNA and survivin protein obviously, and induced apoptosis of HL-60/ADR cells in a time-dependent manner during 12-48 h. After transient transfection with pcDNA3.1/survivin for 48 h, survivin mRNA decreased by 67 %, survivin protein decreased by 57 %, caspase-3 activity increased 4.37 times, and the apoptosis rate increased by 4.41 % compared with control. Compared with ADR alone, pcDNA3.1/survivin significantly reversed MDR in HL-60/ADR cells, the chemosensitivity of HL-60/ADR cells to ADR was increased to 5.36 folds.

CONCLUSION

pcDNA3.1/survivin down-regulated the expression level of survivin mRNA and survivin protein obviously, the threshold of apoptosis was decreased and MDR was reversed.

摘要

目的

研究反义RNA真核表达质粒pcDNA3.1/survivin下调survivin mRNA和survivin蛋白表达水平以及逆转阿霉素耐药HL-60/ADR细胞系多药耐药（MDR）的能力。

方法

采用RT-PCR检测survivin mRNA的表达，Western blot检测survivin蛋白的表达。用Phar Mingen比色法试剂盒测定Caspase-3活性。通过流式细胞术评估细胞凋亡情况。采用MTT法检测HL-60/ADR细胞对阿霉素（ADR）的化疗敏感性。

结果

pcDNA3.1/survivin明显下调survivin mRNA和survivin蛋白的表达水平，并在12至48小时内呈时间依赖性诱导HL-60/ADR细胞凋亡。用pcDNA3.1/survivin瞬时转染48小时后，与对照组相比，survivin mRNA下降了67%，survivin蛋白下降了57%，Caspase-3活性增加了4.37倍，凋亡率增加了4.41%。与单独使用阿霉素相比，pcDNA3.1/survivin显著逆转了HL-60/ADR细胞的多药耐药性，HL-60/ADR细胞对阿霉素的化疗敏感性提高到了5.36倍。