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目的探索功劳木提取物G3对人乳腺癌MCF-7/ADM细胞多药耐药性(Multidrug resistance,MDR)的逆转效果。方法采用MTT法检测G3及阿霉素(Adriamycin,ADM)对MCF-7/S和MCF-7/ADM细胞增殖的抑制作用;RT-PCR法检测G3对MCF-7/ADM细胞MDR1基因mRNA转录水平的影响;免疫组化方法检测G3对MDR1基因产物——P糖蛋白(P-glycoprotein,P-gp)表达水平的影响;流式细胞仪分析G3应用前后MCF-7/ADM细胞内罗丹明123(Rhodamine123,Rh123)浓度的变化,以检测G3对P-gp功能的影响。结果10μg·mL-1为G3的无毒剂量;ADM对MCF-7/S和MCF-7/ADM细胞的IC50分别为(0.22±0.08)μg·mL-1和(51.59±0.12)μg·mL-1;无毒剂量G3与ADM联合应用后对MCF-7/S和MCF-7/ADM细胞的IC分别为(0.19±0.09)μg·mL-1和(20.13±0.53)μg·mL-1,此剂量G3使MCF-7/ADM细胞的IC50下降2.56倍,而对MCF-7/S细胞的IC50无明显影响;无毒剂量的G3应用前后,MCF-7/ADM细胞MDR1基因mRNA转录水平和MDR1基因产物——P-gp的表达水平无明显差异;Rh123蓄积试验中无毒剂量的G3应用后使MCF-7/ADM细胞内Rh123浓度由G3应用前的26.09%升高到86.03%。结论G3能有效逆转人乳腺癌MCF-7/ADM细胞的MDR;G3通过抑制MDR1基因产物——P-gp的“药泵”功能,增加耐药细胞内化疗药物浓度,从而逆转肿瘤细胞的MDR。
Objective To explore the reversal effects of Gokumin K3 extract on multidrug resistance (MDR) in human breast cancer cell line MCF-7 / ADM. Methods MTT assay was used to detect the inhibitory effect of G3 and Adriamycin on the proliferation of MCF-7 / S and MCF-7 / ADM cells. The transcription of MDR1 mRNA was detected by RT-PCR in MCF-7 / ADM cells The effect of G3 on the expression of MDR1 gene product-P-glycoprotein (P-glycoprotein) was detected by immunohistochemistry. Flow cytometry analysis of intracellular rhodamine 123 (Rhodamine123, Rh123) concentration changes in order to test G3 P-gp function. Results The IC50 of ADM on MCF-7 / S and MCF-7 / ADM cells were (0.22 ± 0.08) μg · mL-1 and (51.59 ± 0.12) μg · mL -1; IC of MC3-F7 and MCF-7 / ADM cells were (0.19 ± 0.09) μg · mL-1 and (20.13 ± 0.53) μg · mL-1 The IC50 of MCF-7 / ADM cells decreased 2.56-fold at this dose of G3, but had no significant effect on the IC50 of MCF-7 / S cells. The mRNA transcription level of MDR1 in MCF-7 / ADM cells And the expression level of MDR1 gene -P-gp had no significant difference. Rh123 accumulation increased the Rh123 concentration in MCF-7 / ADM cells from 26.09% before G3 application to 86.03% after application of non-toxic G3. Conclusion G3 can effectively reverse the MDR of human breast cancer MCF-7 / ADM cells. G3 can reverse the tumor cells by increasing the concentration of drug-resistant intracellular chemotherapy by inhibiting the function of MDR1 gene product “P-gp” MDR.