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目的研究姜黄素(curcumin,Cur)及红霉素(erythromycin,EM)对多药耐药(MDR)细胞株 K562/A02的影响及作用机制。方法 MTT 法测定 Cur、EM 作用后 K562/A02细胞对阿霉素(ADM)敏感性的变化。流式细胞仪测定细胞内柔红霉素的平均荧光强度(DNR MFI)。免疫组化法检测细胞膜上 P-gp 的表达。RT-PCR 法检测细胞 mdrl mRNA 水平。结果 Cur、EM 均可减低 ADM 对K562/A02细胞的 IC_(50)值,两药合用时逆转倍数可达11.3倍。K562/A02细胞内 DNR MFI 明显低于K562细胞(P<0.01),Cur、EM 均可明显增加 K562/A02细胞内 DNR MFI(P<0.05),以两药合用时作用最为明显,Cur 2.5μg/ml 处理组细胞内 DNR MFI 略高于 EM 120μg/ml 处理组,但差异无统计学意义(P>0.05)。免疫组化检测结果显示 K562/A02细胞 P-gp 表达明显高于 K562细胞(P<0.01),各组药物分别处理后,K562/A02细胞膜 P-gp 表达减低(P<0.01),但仍高于 K562细胞(P<0.01);各药物组处理5d 细胞膜 P-gp 表达均低于3d 组(P<0.01),Cur 与 EM 合用时细胞膜 P-gp 表达降低最为明显,低于其它处理组(P<0.01)。RT-PCR 结果显示 K562/A02细胞 mdrl mRNA 水平明显高于K562细胞(P<0.01),各组药物处理后,K562/A02细胞 mdrl mRNA 水平均减低(P<0.01),5d 组低于3d 组,但仍高于 K562细胞(P<0.01);Cur 与 EM 合用时 K562/A02细胞 mdrl mRNA 水平降低最为显著,Cur 2.5μg/ml 处理5d K562/A02细胞 mdrl mRNA 水平低于 EM 120 μg/ml 处理5d 组(P<0.01)。结论 Cur、EM 均可部分逆转 K562/A02细胞的 MDR,降低其 P-gp 的表达和功能,逆转作用有时间依赖性;两药联合应用时逆转作用明显增强,Cur 2.5μg/ml 逆转作用略强于 EM 120μg/ml。
Objective To investigate the effect of curcumin (Cur) and erythromycin (EM) on multidrug resistance (MDR) cell line K562 / A02 and its mechanism. Methods The sensitivity of adriamycin (ADM) to K562 / A02 cells was determined by MTT assay. Flow cytometry was used to determine intracellular daunorubicin mean fluorescence intensity (DNR MFI). Immunohistochemistry was used to detect the expression of P-gp on the cell membrane. The mRNA level of mdrl was detected by RT-PCR. Results Both Cur and EM decreased the IC 50 value of ADM on K562 / A02 cells, and the reversal multiple of the two drugs reached 11.3 times. The MFI of DNR in K562 / A02 cells was significantly lower than that in K562 cells (P <0.01). Both Cur and EM significantly increased the DNR MFI of K562 / A02 cells (P <0.05) / ml treatment group intracellular DNR MFI slightly higher than EM 120μg / ml treatment group, but the difference was not statistically significant (P> 0.05). The results of immunohistochemistry showed that the expression of P-gp in K562 / A02 cells was significantly higher than that in K562 cells (P <0.01). The P-gp expression in K562 / A02 cells was decreased (P <0.01) (P <0.01). The expression of P-gp in cell membrane of all drug groups was lower than that of 3d group (P <0.01) P <0.01). The mRNA level of mdrl in K562 / A02 cells was significantly higher than that in K562 cells (P <0.01) by RT-PCR. The mRNA levels of mdr1 in K562 / A02 cells were decreased after treatment (P <0.01) , But still higher than that of K562 cells (P <0.01). The mdrl mRNA level of K562 / A02 cells was most significantly decreased when Cur was combined with EM, and mdrl mRNA level of K562 / A02 cells was lower than that of EM 120 μg / ml Treatment 5d group (P <0.01). Conclusions Both Cur and EM can partially reverse the MDR of K562 / A02 cells and decrease the expression and function of P-gp, and the reversal effect is time-dependent. The reversal effect of both drugs is obviously enhanced, and the reversal effect of Cur 2.5μg / ml is slightly Stronger than EM 120 μg / ml.