摘要
AIM To filtrate breast cancer resistance protein(BCRP)-mediated resistance agents and investigatethe mechanism,so as to provide valuable datum for optimization clinical chemotherapy scheme to tumor withevaluation marker of BCRP expression.METHODS MTT assay was used to filtrate BCRP-mediatedresistance agents with PA317/Tet-on/TRE-BCRP cell of different expression levels of BCRP after treated withdifferent concentration anticancer agents.High performance liquid chromatography(HPLC) was applied tomeasure relative dose of intracellular retention resistance agents.Nuclear DNA fluorescence dye,Hochest33258,staining and flow cytometry were adopted to detect apoptotic cells after treated with drugs.RESULTSThere were shown increasing durg-resistance to 5-fluorouracil,methotrexate,doxirubicin,pirarubicin,etoposide and mitoxantrone followed with increasing expression of BCRP on PA317/Tet-on/TRE-BCRPcells(P<0.05,n=3),but shown sensitive to paclitaxel,cisplatin,vincristine,mitomycin and vindesine.Therealso was shown significant negative correlation between the intracellular retention dose of 5-fluorouracil withdifferent expression of BCRP(r=-0.885,P<0.05,n=3).There were shown parallel results ofthat decreasingcellular apoptotic rate with increasing cellular expression of BCRP after treated with 5-fluorouracil byfluorescence dye staining and flow cytometry(P<0.05,n=3),and also shown significate rise of the apoptoticrate of BCRP expression cells after treated with Ko143 (P<0.05,n=3).Every group of cells could be differentextently blocked in phase of G_0/G_1 treated with 5-fluorouracil.CONCLUSION Resistance of 5-fluorouracilcould be especially mediated by conjugated with BCRP and acted as drug exclude-pump substrate.Cellularability resistant to 5-fluorouracil-induced apoptosis could be reinforced by BCRP expression.
AIM To filtrate breast cancer resistance protein（BCRP）-mediated resistance agents and investigate the mechanism,so as to provide valuable datum for optimization clinical chemotherapy scheme to tumor with evaluation marker of BCRP expression. METHODS MTT assay was used to filtrate BCRP-mediated resistance agents with PA317/Tet-on/TRE-BCRP cell of different expression levels of BCRP after treated with different concentration anticancer agents. High performance liquid chromatography（HPLC） was applied to measure relative dose of intracellular retention resistance agents.Nuclear DNA fluorescence dye,Hochest 33258, staining and flow cytometry were adopted to detect apoptotic cells after treated with drugs. RESULTS There were shown increasing durg-resistance to 5-fluorouracil,methotrexate, doxirubicin, pirarubicin, etoposide and mitoxantrone followed with increasing expression of BCRP on PA317/Tet-on/TRE-BCRP cells（P〈0.05, n=3）,but shown sensitive to paclitaxel, cisplatin, vincristine, mitomycin and vindesine. There also was shown significant negative correlation between the intracellular retention dose of 5-fluorouracil with different expression of BCRP（r=-0.885, P〈0.05, n=3）.There were shown parallel results of that decreasing cellular apoptotic rate with increasing cellular expression of BCRP after treated with 5-fluorouracil by fluorescence dye staining and flow cytometry（P〈0.05, n=3）,and also shown significate rise of the apoptotic rate of BCRP expression cells after treated with Ko143 （P〈0.05, n=3）. Every group of cells could be different extently blocked in phase of G0/G1 treated with 5-fluorouracil. CONCLUSION Resistance of 5-fluorouracil could be especially mediated by conjugated with BCRP and acted as drug exclude-pump substrate. Cellular ability resistant to 5-fluorouracil-induced apoptosis could be reinforced by BCRP expression.
出处
《深圳特区科技》
2005年第11期106-112,共7页
Shenzhen Science & Technology
关键词
乳腺癌耐受蛋白
5-氟脲嘧啶
耐受机制
耐药性
breast cancer resistance protein
drug resistance
5-fluorouracil
apoptosis
anticancer drugsResistance of malignant cells to multiple chemotherapeutic agents is a major obstacle to the successfultherapy of cancer, known as multidrug resistance（MDR）.Expression of ATP-binding cassette（ABC）superfamily transporters is one of the predominant mechanisms of MDR.Breast cancer resistanceprotein（BCRP） is one new member of ABC superfamily transporters for the first time reported as 199811].1tsresearch is paid close attention to extremely.Research has indicated already that BCRP perform an atypicalmultidrug resistance phenotype.lt offer some bases for function of BCRP on multidrug resistance in tumor, butfor resistance under independently BCRP, kinds of drug-resistance and function mechanisms,there are stilllack the systematic materials.For to precisely clarificate behavior and law of BCRP-mediate atypicaldrug-resistance,in our study,we utilize successfully established model of expression of BCRP withdoxycycline（DCL） induced Tet regulating system in mouse fibroblast cell line（PA317） by ourselves[2] toobserve effect-dose relation between degree of drug-resistance and expression level of BCRP and filtrateBCRP-mediate resistance agents and investigate its mechanism,so as to provide valuable datum foroptimization clinical chemotherapy scheme to tumor with evaluation marker of BCRP expression.
