三七总皂苷逆转乳腺癌细胞MCF-7-ADM多药耐药实验探究.docVIP

三七总皂苷逆转乳腺癌细胞MCF-7/ADM多药耐药实验探究 　 作者：刘丽丽， 刘艳娥，房国涛 【摘要】 　　目的从中药中寻找安全有效的多药耐药逆转剂,并初步研究三七总皂苷(PNS) 对人乳腺癌细胞MCF-7/S(敏感细胞) 及MCF-7/ADM(耐药细胞) 的影响。方法以MTT 法测定PNS对MCF-7/S 及MCF-7/ADM的直接细胞毒作用；测定阿霉素(DOX) 对两种细胞的毒性作用并计算出耐药倍数；以无细胞毒性的三七总皂苷注射液作为耐药逆转剂用MTT法体外药敏感实验观察其对多药耐药细胞株MCF-7/ADM的逆转作用；用RT-PCR分别测敏感细胞和PNS作用后的耐药细胞表达的MDR1基因的阳性率；用WESTERN测定敏感细胞和PNS作用后的耐药细胞表达多药耐药蛋白P - gp (P - 170) 的阳性率。以上实验均用维拉帕米作为阳性对照。结果①PNS在300μg/ ml 浓度以下时对两细胞基本无毒性,为安全有效的逆转剂量。②阿霉素对敏感细胞和耐药细胞的半数抑制浓度( IC50) 分别为0.62μg/ ml 和25.03 μg/ ml，耐药倍数为40.37倍。③PNS能够增强阿霉素(DOX) 对MCF-7/ADM的细胞毒作用, PNS 50，100，200μg/ ml分别作用于乳腺癌耐药细胞48 h后，耐药细胞株的IC50分别降至17.85，13.80，11.63 μg/ml。④PNS(50，100，200μg/ ml) 均可下调多药耐药基因mdr - 1 及其表达的糖蛋白P - gp 的量,且随PNS浓度的增加，下调作用更加明显。而未发现维拉帕米有类似作用。结论①高浓度的三七总皂苷具有一定的抗肿瘤作用；②三七总皂苷可下调MCF-7/ADM的MDR-1和P-gP；③PNS能够部分逆转耐药细胞MCF-7/ADM的多药耐药性；④三七总皂苷是一种有效安全的多药耐药逆转剂。 【关键词】 乳腺癌 三七总皂苷 多药耐药 逆转 体外研究 　　Abstract：ObjectiveTo search a sort of safe and effective reversal agent for multidrug resistantce from Chinese traditional medicine and to study the impact of the Panaxnotoginseng saponins (PNS) on human breast clone MCF-7/S and multidrug resistance clone MCF-7/ADM in vitro. MethodsTo measure direct cell toxicant effect of PNS to MCF-7/S(sensitive strain) and MCF-7/ADM (multidrug-resistant strain) and measure toxicant effect of dox to both strains for resistant index by MTT;to observe the reversal effect of PNS which has screened for nontoxicant concentration to MCF-7/ADM by MTT;to measure expression positive rate of mdr-1 and P-gp of the both cell after treatment with PNS by PCR and WESTERN .Veraparmil was used as a positive control.Results①There was no obvious cell toxicity of PNS to MCF-7/S and MCF-7/ADM cell,when the concentration of PNS was 300 μg/ml below. ②IC50 of Dox to MCF-7/S and MCF-7/ADM were 0.62 ng/ml and 25.03 ng/ml respectively.Drug-Resistant index of MCF-7/ADM was 40.37.③PNS could increase cell toxic effect of dox,after treated with could reduce manner 50，100，200 μg/ml PNS respectively to MCF-7/ADM for 48 hours,IC50 respectively decl