解毒祛瘀方对乳腺癌细胞MCF-7/ADM的耐药逆转作用

目的:研究解毒祛瘀方对人乳腺癌耐药细胞MCF-7/ADM耐药逆转的作用及机制。方法:以MCF-7/ADM细胞为研究对象,利用噻唑蓝(MTT)比色法检测解毒祛瘀方对人乳腺癌耐药细胞MCF-7/ADM生长的影响;应用流式细胞术检测肿瘤细胞内罗丹明123(Rh-123)的含量;分别利用实时荧光定量聚合酶链式反应(Real-time PCR)及蛋白免疫印迹法(Western blot)检测肿瘤细胞内多药耐药蛋白1(MDR1),乳腺癌耐药相关蛋白(BCRP)mRNA及蛋白表达变化。结果:与空白组比较,经1.25,2.5 g·L~(-1)解毒祛瘀方作用后,阿霉素对人乳腺癌耐药细胞MCF-7/ADM的逆转倍数(RF)分别提升1.7倍和3.0倍(P0.05);人乳腺癌耐药细胞MCF-7/ADM中Rh-123含量分别提高了1.8倍和2.5倍(P0.05),MDR1和BCRP蛋白和mRNA表达水平明显下降(P0.05),1.25,2.5 g·L~(-1)解毒祛瘀方MDR1 mRNA表达分别降低35.5%和56.0%(P0.05),BCRP mRNA表达分别降低41.6%和49.5%(P0.05)。结论:解毒祛瘀方可提高人乳腺癌耐药细胞MCF-7/ADM对阿霉素的敏感性,逆转该细胞对阿霉素的耐药性,其机制可能与降低MDR1和BCRP蛋白和mRNA的表达,抑制细胞药物外排作用相关。     

温下方逆转MCF-7/ADM细胞耐药及诱导凋亡作用

目的:研究自拟温下方逆转MCF-7/ADM细胞耐药、诱导凋亡作用.方法:MTT法研究温下方含药血清对MCF-7/ADM耐药逆转作用,流式细胞术测定细胞内药物浓度及耐药相关蛋白P-gp、凋亡调控蛋白p53、bcl-2的表达,激光共聚焦显微镜技术检测细胞内钙浓度及线粒体膜电位.结果:温下方含药血清可提高MCF-7/ADM对化疗药的敏感性,增加胞内化疗药物浓度,使P-gp表达下降,p53表达增高,胞内钙浓度及线粒体膜电位均降低.结论:温下方可部分逆转MCF-7/ADM耐药,其逆转机制可能与诱导耐药细胞凋亡密切相关.     

补骨脂素逆转人乳腺癌耐药细胞株MCF-7多药耐药性研究

人乳腺癌细胞产生多药耐药性(multidrugresistance,MDR)是肿瘤化疗失败的主要原因之一。已知某些中药具有钙拮抗作用,且药物作用较温和,能逆转MDR从而提高化疗效果,具有很好的应用和开发前景。补骨脂是补益药中少有的钙离子通道拮抗剂,本实验选用其主要成分之一的补骨脂素为研究对象,进行逆转人乳腺癌细胞株耐药性的研究,为今后进一步开发有效耐药逆转剂打下基础。材料和方法1·材料实验用细胞株MCF-7(人乳腺癌细胞株)及MCF-7/ADR(用长春新碱诱导的MCF-7多药耐药细胞株)由北京大学提供。RPMI-1640由GIBCO公司出品,P-170单抗购于…     

补骨脂素逆转人乳腺癌耐药细胞MCF-7／ADR多药耐药性的研究

目的研究具有钙拮抗作用的中药补骨脂素对多耐药性的逆转作用。方法：选用异搏定作阳性对照，观察其对多药耐药细胞系MCF-7／ADR多药耐药性的逆转作用。结果：补骨脂素在非细胞毒性剂量下能使MCF-7／ADR对阿霉素的浓度升高。结论：补骨脂素具有逆转人乳腺癌MCF-7／ADR多药耐药性的作用。     

漏芦抽提剂对人乳腺癌耐药细胞MCF-7/ADR的耐药逆转作用研究

目的:研究漏芦抽提剂逆转肿瘤耐药的作用。方法:用漏芦抽提剂及其含药血清处理人乳腺癌耐药细胞株(MCF-7/ADR),以MTT法测定漏芦抽提剂及其含药血清对MCF-7/ADR细胞系的耐药逆转作用。结果:漏芦抽提剂对MCF-7/ADR细胞系具有很强的细胞毒作用,与ADM合用培养96 h细胞死亡率平均为ADM组的1.63倍,与耐药逆转剂维拉帕米(VRP)相比(ADM+VRP组为ADM的1.78倍),无显著性差异(P>0.05)。漏芦抽提剂含药血清(大剂量)和ADM合用时,培养96 h细胞死亡率平均为ADM的1.70倍,这一结果与耐药逆转剂维拉帕米作用结果相似。结论:漏芦抽提剂在逆转肿瘤多药耐药方面可能具有良好的应用前景。     

中药逆转MCF-7/ADM细胞多药耐药机制的研究进展

乳腺癌细胞(MCF-7/ADM)的多药耐药性(MDR)是导致抗肿瘤药物临床化疗失败的主要原因。现认为细胞中的P-gp、Bcl-2、GST-π、TopoⅡ等蛋白可能与乳腺癌的多药耐药相关。近年来研究发现,不少中药能够下调MDR1 mRNA和/或P-gp的表达,从而逆转MCF-7/ADM细胞对抗肿瘤药物的多药耐药性,提高耐药细胞对化疗药物敏感性。主要对MCF-7/ADM细胞的多药耐药机制及有逆转MCF-7/ADM多药耐药作用的中药进行了综述。     

中药逆转乳腺癌多药耐药的研究概况及展望

综述近年中药逆转乳腺癌多药耐药的研究概况。目前中药逆转乳腺癌多药耐药研究较多的中药单体及提取物有姜黄、人参、苦参等,中药复方有复方紫龙金、逆转胶囊、疏肝益肾方等;复方研究少、体内研究少、机制探讨不够深入是目前中药逆转乳腺癌多药耐药面临的主要问题。     

中药功劳木对乳腺癌耐药MCF7/ADM逆转MDR1的作用研究

目的：研究中药功劳木对乳腺癌耐药MCF7/ADM逆转MDR1的作用。方法：应用台盼蓝拒染试验测试应用功劳木后ADM对MCF7/ADM的活细胞率：MTT法测试功劳木的细胞毒性及对ADM的增效;Rh123荧光技术检测应用功劳木对耐药细胞内药物浓度的影响,流式细胞仪检测功劳木对MDR1的影响。结果：实验组浓度为10 mg/L以及20mg/L时对MCF7/ADM的抑制分别为（0.218±0.081）,（0.202±0.033）,均明显高于对照组（0.146±0.062）,（0.153±0.018）,差异有统计学意义（P〈0.05）。实验组对MCF-7/S以及MCF7/ADM的抑制分别为（0.204±0.076）,（51.08±0.63）,明显高于对照组的（0.075±0.032）及（36.55±0.53）,差异有统计学意义（P〈0.05）。结论：GLM对乳腺癌耐药MCF7/ADM逆转MDR1具有抑制作用。     

中药逆转乳腺癌细胞多药耐药性的实验研究

目的:从单味中药中筛选具有逆转乳腺癌细胞耐药性的逆转剂.方法:建立乳腺癌耐药细胞的细胞模型,采用SRB法测定单味中药及其对应血清的细胞毒性作用及其逆转作用.结果:理气类中药中延胡索具有对MCF-7/VCR耐药性的逆转作用,而且它的细胞毒作用很小;1.667g/L的延胡索可使MCF-7/VCR的耐药性逆转3.27倍.结论:延胡索能够有效地逆转乳腺癌细胞MCF-7/VCR的耐药性.     

硒化壳聚糖逆转K562/ADM细胞多药耐药的实验研究

目的:研究硒化壳聚糖对人慢性粒细胞白血病耐阿霉素细胞株( K562/ADM) mdr-1基因/P-糖蛋白(P-gp)表达及功能的影响,为逆转肿瘤多药耐药提供新的途径.方法:硒化壳聚糖100,200 mg·L-1作用K562/ADM细胞24 h,应用RTPCR法和免疫印迹法检测mdr-1/P-gp表达的改变;应用高效液相色谱法检测细胞内阿霉素积聚浓度;应用MTT法检测阿霉素对K562/ADM细胞增殖的影响.结果:硒化壳聚糖可增强K562/ADM细胞对阿霉素的敏感性,增加细胞内阿霉素集聚浓度,200 mg·L-1硒化壳聚糖作用效果显著强于100 mg·L-1硒化壳聚糖(P＜0.01);硒化壳聚糖能明显抑制K562/ADM细胞mdr-1/P-gp的表达(P＜0.01),100 mg·L-1硒化壳聚糖可使mdr-1/P-gp表达分别下降(40.87 -3.19)％和(35.08±0.09)％,200 mg·L-1硒化壳聚糖可使mdr-1/P-gp表达分别下降(78.24±3.42)％和(79.61±0.23)％.结论:硒化壳聚糖可明显抑制K562/ADM细胞mdr-1/P-gp表达,增加细胞内阿霉素含量,恢复细胞对化疗药物敏感性,逆转mdr-1编码蛋白P-gp介导的多药耐药.     

五味子颗粒辅助CAF化疗方案对乳腺癌多药耐药的逆转作用

目的：观察五味子颗粒辅助联合环磷酰胺+阿霉素+5-氟尿嘧啶（CAF）化疗方案对乳腺癌多药耐药的逆转作用及对多药耐药相关蛋白（MRP）的影响。方法：72例MRP阳性表达乳腺癌患者按住院前后顺序分为实验组和对照组各36例。对照组采用CAF化疗方案进行治疗,实验组在对照组治疗的基础上给予五味子颗粒辅助治疗。两组疗程均为8周。观察两组的临床疗效、生存时间、中位缓解期、治疗后副作用及逆转多药耐药相关蛋白（MRP）的有效率。结果：实验组总有效率为58.33%,明显优于对照组的30.56%（P<0.05）;实验组中位缓解期为12.5个月,明显好于对照组的7.5个月（P<0.05）;治疗后实验组逆转MRP有效率为33.3%,明显高于对照组的2.78%（P<0.05）。结论：五味子颗粒联合CAF化疗方案可以有效缓解乳腺癌患者的临床症状,可有效逆转MRP介导的肿瘤多耐药性,在肿瘤治疗中发挥一定作用。     

15-PGDH�յ�ҩ����ת�����ٰ�MCF-7/ADRϸ����ҩ���ü�����

??OBJECTIVE To examine the expression of 15-hydroxyprostaglandin dehydrogenase(15-PGDH) in human multidrug-resistant breast cancer line MCF-7/ADR and to explore the reversal effect and mechanism of 15-PGHD induction drugs on MCF-7/ADR cells. METHODS The RT-PCR and Western blot were used to detect 15-PGDH, COX-2 mRNA and protein expression in MCF-7 and MCF-7/ADR cells. PGE₂ levels in supernatant of cells were determined by ELISA assay. Anti-proliferation effect and chemotherapy sensitivity to ADM of 15-PGDH induction drugs (indomethacin, ibuprofen and pioglitazone, dexamethasone) on breast cancer cells were assayed by MTT method. Cell apoptosis was detected by Hochest 33258 stain assay. RESULTS Compared with MCF-7 cells, the 15-PGDH expression was significantly decreased, COX-2 expression was significantly increased and PGE₂ levels in cell supernatant were increased in MCF-7/ADR cells. 15-PGDH induction drugs (indomethacin, ibuprofen and pioglitazone, dexamethasone) increased 15-PGDH expression or both reduced COX-2 expression, and finally reduced PGE₂ levels in MCF-7/ADR cells. Effect of chemosensitivity and apoptosis induction of ADM was enhanced and multidrug resistance was partially reversed when co-treated with 15-PGDH induction drugs. CONCLUSION The expression of 15-PGDH is decreased in human multidrug-resistant breast cancer line MCF-7/ADR. 15-PGDH induction drugs could increase chemosensitivity, promote apoptosis and reverse resistance of MCF-7/ADR cell, the mechanism might related to the influence of PGE₂ level by regulated the expression of 15-PGDH and COX-2.     

�����������������������������ת���ٰ���ҩ��ҩ����������

??OBJECTIVE To prepare a redox and pH dual sensitive nano-carrier based on PAMAM in order to co-loading chemotherapeutics doxorubicin and breast cancer multidrug resistance reversal agent elacridar, and study their in vitro reversal effect. METHODS The infrared spectrum FTIR was used to characterize the carrier. Confocal was used to investigate the intracellular triggered drug release. The reversal effect of breast cancer multidrug resistance and the in vitro anti-tumor activity of doxorubicin and elacridar co-loaded nanoparticles were investigated using flow cytometry and cell toxicity tests, respectively. RESULTS The doxorubicin and elacridar co-loaded nanoparticles (PSSP/DOX/ELC) were successfully prepared, and pH-redox dual sensitive of carrier was proved by cell experiments.And the carrier was uptaken into cells and delivery to lysosome, and drug release was triggered in the lysosome acid condition, then the released drug diffused to the nucleus. The trial of rhodamine 123 accumulation and efflux assay revealed that the accumulation of rhodamine 123 was notably increased after incubation of elacridar in MCF-7/ADR cells. The cytotoxicity of PSSP/DOX/ELC nanoparticles against MCF-7/ADR cell line was significantly stronger than that of either free doxorubicin or only doxorubicin loaded nanoparticles (PSSP/DOX). CONCLUSION The reversal effect of multidrug resistance and the cytotoxicity of cancer cells were significantly enhanced by PSSP/DOX/ELC nanoparticles. PSSP/DOX/ELC nanoparticles is a promising delivery system.     

��ҩ��ҩ1����С����RNA������֬������ת���ٰ���ҩ��ҩ���о�

??OBJECTIVE To explore the feasibility of the reversal of multidrug resistance in breast cancer using vector-based small interference RNA(siRNA) and to solve the problems of siRNA transient expression and siRNA delivery in vivo. METHODS Based on the siRNA sequence which was screened in previous studies that could effectively inhibit the expression of MDR1 gene, the expression plasmid was constructed. The siRNA expression plasmid was then encapsulated in new nano-sized stealth cationic liposomes. Pharmacodynamic studies of the liposomes were carried out in vitro and in vivo. RESULTS The results showed that the cationic liposomes loaded with MDR1 siRNA expression plasmid could effectively inhibit the expression of MDR1 gene both in vitro and in vivo. CONCLUSION Multi-drug resistance of breast cancer cells is reversed to a great extent by this siRNA-containing cationic liposomes. Nano-sized cationic liposomes are ideal delivery vehicle of siRNA, which could protect the siRNA from degradation and deliver siRNA into the tumor region where it could exert functions.
     

复方藤梨根制剂对K562/ADM多药耐药作用的体内实验研究

目的：研究复方藤梨根制剂逆转人红白血病耐药细胞株K562/ADM裸鼠移植瘤的多药耐药效应及机制。方法：建立K562/ADM裸鼠移植瘤多药耐药模型;以流式细胞仪（FCM）检测各组肿瘤细胞膜上P-gp的表达和肿瘤细胞内ADM蓄积浓度。结果：（1）ADM组P-gp的表达与NS组相比明显升高（P（0.05）;不同剂量的FFTLG制剂与ADM合用时,P-gp的表达与ADM组相比显著下降（P（0.01）。单独应用FFTLG制剂,当浓度为0.8mg/mL时P-gp的表达与ADM组相比下降（P（0.05）。（2）与ADM组相比,ADM和不同剂量的FFTLG合用组肿瘤细胞内的ADM浓度均有上升（P（0.05）。结论：FFTLG制剂可以部分逆转荷瘤裸鼠的多药耐药性,其逆转效果似呈剂量依赖性。FFTLG制剂逆转荷瘤裸鼠多药耐药性的机理之一是通过下调肿瘤细胞膜上P-gp的表达或抑制P-gp的活性,增加肿瘤细胞内ADM的蓄积浓度而实现的。     

硒化壳聚糖对K562/ADM细胞生物学行为的影响

目的 观察硒化壳聚糖对体外培养慢性粒细胞白血病多药耐药细胞株K562/ADM细胞生物学行为的影响.方法 硒化壳聚糖作用K562/ADM细胞12～24 h,应用流式细胞法检测细胞凋亡,软件拟合计算细胞周期;应用免疫印迹法检测P-gp的表达;应用RT-PCR法检测mdr-1 mRNA水平.结果 硒化壳聚糖能够明显增强ADM对K562/ADM细胞的诱导凋亡作用,阻滞细胞周期于G1期(P＜0.05,P＜0.01),下调P-gp表达和mdr-1mRNA水平(P ＜0.05,P＜0.01),硒化壳聚糖浓度越高,作用效果越明显.结论 硒化壳聚糖能够通过下调mdr-1基因和P-gp蛋白表达,阻滞细胞于G1期诱导凋亡来对体外培养的K562/ADM细胞耐药生物学行为产生影响.