[生存素反义寡脱氧核苷酸对K562细胞耐药性的影响]

[Effects of survivin antisense oligodeoxynucleotide on drug resistance in K562 cells].

作者信息

Yang Shan, Xu You-hua, Wen Xian-hao, Chen Lin, Li Xin

机构信息

Department of Hematology, Children's Hospital of Chongqing Medical University, Chongqing 400014, China.

出版信息

Sichuan Da Xue Xue Bao Yi Xue Ban. 2008 Nov;39(6):905-8.

PMID:19253822

Abstract

OBJECTIVE

To study the effect of Survivin antisense oligodeoxgnucleotide (ASODN) on drug resistance and its mechanism in K562 cells.

METHODS

Cells were divided into four groups: ASODN group, SODN group, Lip group and blank group. Survivin ASODN was transfered into K562 cells by liposomal reagent. The expression of Survivin mRNA was detected by RT-PCR. The sensitivity of K562 cells to adriamycin (ADM) and daunorubicin (DAM) was detected by MTT assay, while the intracellular concentration of ADM and DAM was measured by flowcytometry.

RESULTS

Compared to control group Survivin mRNA expression in ASODN group decreased obviously. IC50(s) of ADM and ASODN+ADM were 0.5457 mg/L and 0.1933 mg/L respectively, IC50 (s) of DAM and ASODN+DAM were 0.5408 mg/L and 0.2027 mg/L respectively. Expression of Survivin mRNA decreased by 25.8% after the transfection of ASODN. Fluorescence intensity of ADM and ASODN+ADM in K562 cells were 51.64 and 89.92, Fluorescence intensity of DAM and ASODN+DAM in K562 cells were 63.71 and 88.47.

CONCLUSION

Expression of Survivin mRNA in k562 cells was down-regulated by ASODN. Survivin ASODN is able to reverse the drug resistance via inhibition of Survivin expression and inducement of apoptosis.

摘要

目的

研究生存素反义寡脱氧核苷酸（ASODN）对K562细胞耐药性的影响及其机制。

方法

将细胞分为四组：ASODN组、SODN组、Lip组和空白组。通过脂质体试剂将生存素ASODN转入K562细胞。采用RT-PCR检测生存素mRNA的表达。采用MTT法检测K562细胞对阿霉素（ADM）和柔红霉素（DAM）的敏感性，同时采用流式细胞术检测细胞内ADM和DAM的浓度。

结果

与对照组相比，ASODN组生存素mRNA表达明显降低。ADM和ASODN+ADM的IC50分别为0.5457mg/L和0.1933mg/L，DAM和ASODN+DAM的IC50分别为0.5408mg/L和0.2027mg/L。转染ASODN后生存素mRNA表达下降25.8%。K562细胞中ADM和ASODN+ADM的荧光强度分别为51.64和89.92，K562细胞中DAM和ASODN+DAM的荧光强度分别为63.71和88.47。