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目的研究中药川芎嗪(TMP)与环孢菌素A(CsA)联合应用对转基因多药耐药细胞K562/MDR的耐药性逆转作用及可能机制。方法应用非细胞毒性剂量的TMP和CsA单独及联合作用于K562/S和K562/MDR细胞,MTT法检测细胞对阿霉素(ADM)的药敏性,计算耐药倍数及逆转倍数。利用罗丹明123(Rh123)进行蓄积与排出试验,免疫组化检测P-gp的表达,RT-PCR方法检测MDR1基因的表达。结果与K562/S细胞相比,K562/MDR细胞的耐药倍数为19.2倍;TMP及CsA可显著降低ADM对K562/MDR细胞的IC50(P<0.01),逆转倍数分别为4.9倍及9.2倍,两者联合应用,逆转倍数为15.4倍。TMP及CsA单独应用均能降低K562/MDR细胞P-gp的表达,联合应用使K562/MDR细胞P-gp的表达率明显低于单独应用(P<0.01);TMP和CsA单独及联合应用,MDR1基因表达均有不同程度的下调。结论 TMP和CsA联合逆转K562/MDR细胞的多药耐药性具有协同作用,其机制可能与下调MDR1基因表达和降低P-gp水平有关
Objective To investigate the reversal effect and possible mechanism of drug resistance of multi-drug resistant cell line K562 / MDR combined with tetramethylpyrazine (TMP) and cyclosporine A (CsA). Methods K562 / S and K562 / MDR cells were treated with non-cytotoxic doses of TMP and CsA alone or in combination with K562 / S and K562 / MDR cells. MTT assay was used to determine the susceptibility of ADM to ADM. Accumulation and excretion of Rhodamine 123 (Rh123) were performed. The expression of P-gp was detected by immunohistochemistry. The expression of MDR1 gene was detected by RT-PCR. Results Compared with K562 / S cells, the multidrug resistance of K562 / MDR cells was 19.2 fold, and the IC50 of ADM and K562 / MDR cells was significantly decreased by TMP and CsA (P <0.01) , A combination of the two, reversal multiple was 15.4 times. The combination of TMP and CsA could decrease the expression of P-gp in K562 / MDR cells alone. The combined application of P-gp in K562 / MDR cells was significantly lower than that in K562 / MDR alone group (P <0.01) MDR1 gene expression were down to different degrees. Conclusions The combination of TMP and CsA can reverse the multidrug resistance of K562 / MDR cells in a synergistic manner. The mechanism may be related to the down-regulation of MDR1 gene expression and the decrease of P-gp level