姜黄素与红霉素逆转K562/A02细胞系多药耐药的机制研究

Chang Hong-yu, Pan Kai-li, Ma Fu-cheng, Jiao Xi-ying, Zhu Hua-feng, Liu Jian-hong, Huang Ying, Cao Yu-hong

Department of Paediatrics, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, China.

Zhonghua Xue Ye Xue Za Zhi. 2006 Apr;27(4):254-8.

OBJECTIVE

To investigate the effects of curcumin (Cur) and erythromycin (EM) on multidrug resistance (MDR) reversal of K562/A02 cell line and their mechanism.

METHODS

MTT assay was employed to determine the sensitivity of Cur, EM-treated K562/A02 cells to adriamycin (ADM). Flow cytometry was used to measure intracellular mean fluorescence intensity (MFI) of daunorubicin (DNR). P-gp expression was determined by immunohistochemistry. RT-PCR technique was used to examine the mdr1 mRNA level.

RESULTS

IC(50) of ADM in K562/A02 cells was decreased when treated with Cur or EM, and the reversal times (RvT) was 4.9, 3.7 respectively. The RvT reached to 11.3 when treated with Cur (2.5 microg/ml) combined with EM (120 microg/ml). The DNR MFI in K562/A02 cells was significantly lower than that in K562 cells (P < 0.01), and was increased significantly when treated with Cur (2.5 microg/ml) or EM (120 microg/ml) (P < 0.05). There was no significant difference between DNR MFI of K562/A02 cells treated with Cur (2.5 microg/ml) or EM (120 microg/ml). Immunohistochemistry showed that P-gp expression was significantly higher in K562/A02 cells than in K562 cells (P < 0.01), and was reduced in K562/A02 cells treated with each (P < 0.01), though being still higher than that in K562 cells (P < 0.01). P-gp expression of K562/A02 cells treated with each drug for 5 days were lower than that for 3 days (P < 0.01), and lowered further when treated with Cur and EM together (P < 0.01). Mdr1 mRNA level in K562/A02 cells was higher than in K562 cells (P < 0.01), and was decreased when treated with each of the drugs (P < 0.01). The mdr1 mRNA level of K562/A02 cells treated with Cur (2.5 microg/ml) plus EM (120 microg/ml) was decreased most significantly than that treated with other group of drugs. After 5 day treatment the mdr1 mRNA level of K562/A02 cells with Cur (2.5 microg/ml) was lower than that with EM 120 microg/ml (P < 0.01).

CONCLUSIONS

Either Cur or EM can partly reverse the multidrug resistance of K562/A02 cells and decrease the expression and function of P-gp in a time-dependent way. MDR reversing effect of Cur combined with EM is stronger than that of Cur or EM alone.

目的

探讨姜黄素（Cur）和红霉素（EM）对K562/A02细胞系多药耐药（MDR）逆转作用及其机制。

方法

采用MTT法检测Cur、EM作用后K562/A02细胞对阿霉素（ADM）的敏感性。流式细胞术检测柔红霉素（DNR）细胞内平均荧光强度（MFI）。免疫组化法检测P-糖蛋白（P-gp）表达。RT-PCR技术检测mdr1 mRNA水平。

结果

Cur或EM作用后K562/A02细胞中ADM的半数抑制浓度（IC50）降低，逆转倍数（RvT）分别为4.9、3.7。Cur（2.5μg/ml）联合EM（120μg/ml）作用时RvT达11.3。K562/A02细胞中DNR的MFI显著低于K562细胞（P<0.01），Cur（2.5μg/ml）或EM（120μg/ml）作用后显著升高（P<0.05）。Cur（2.5μg/ml）与EM（120μg/ml）作用后K562/A02细胞中DNR的MFI差异无统计学意义。免疫组化显示，K562/A02细胞中P-gp表达显著高于K562细胞（P<0.01），各药物作用后K562/A02细胞中P-gp表达均降低（P<0.01），但仍高于K562细胞（P<0.01）。各药物作用K562/A02细胞5天时P-gp表达低于作用3天时（P<0.01），Cur与EM联合作用时降低更明显（P<0.01）。K562/A02细胞中mdr1 mRNA水平高于K562细胞（P<0.01），各药物作用后均降低（P<0.01）。Cur（2.5μg/ml）加EM（120μg/ml）作用后K562/A02细胞中mdr1 mRNA水平降低最明显。Cur（2.5μg/ml）作用5天后K562/A02细胞中mdr1 mRNA水平低于EM（120μg/ml）（P<0.01）。