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ObjectiveTo explore the effect of Gelsemium elegans combined with Mussaenda pubescens on efflux transporter breast cancer resistance protein （BCRP） and cytochrome P450 3A11 （CYP3A11） and their attenuation mechanism， and to investigate whether the nuclear receptors were involved in such regulation by intervening it with nuclear receptor activators. MethodC57BL/6 mice were divided into the blank group， G. elegans （GE， 0.25 g·kg-1）group， GE + M. pubescens （MP） （0.25 g·kg-1+10 g·kg-1） group， GE + pregnane X receptor （PXR） activator （rifampicin）（GE + Rif，0.25 g·kg-1+50 mg·kg-1） group， GE + MP + Rif （0.25 g·kg-1+10 g·kg-1+50 mg·kg-1） group， GE + constitutive androstane receptor （CAR） activator （1，4-Bis ［2-（3，5-Dichloropyridyloxy）］ benzene， TCPOBOP）（GE + TCP， 0.25 g·kg-1+0.5 mg·kg-1） group， and GE + MP + TCP （0.25 g·kg-1+10 g·kg-1+0.5 mg·kg-1） group. The medication lasted for 14 successive days. One hour after the last administration， the mice were sacrificed by cervical dislocation and the liver tissue was harvested. The left liver tissue was stained with hematoxylin- eosin （HE） for observing the pathological changes. The right liver tissue was used for BCRP and CYP3A11 mRNA and protein expression detection by real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot. ResultThe survival rates of mice in the GE + Rif group， GE group， and GE + MP group were 25% （the lowest）， 40%， and 80%， respectively， and no death was observed in the other groups. Compared with the obvious lesions in the liver cells of the GE group， the pathological changes in liver cells of the GE + MP group were alleviated， while those in the GE + Rif group were worsened. Compared with the GE + Rif group， the GE + MP + Rif group exhibited relieved pathological changes in liver cells. Both the GE + TCP group and the GE + MP + TCP group showed mild liver lesions. The comparison with the GE + MP group revealed that the pathological changes in the GE + MP + TCP group were slightly relieved. Compared with the blank group， the expression of BCRP protein and mRNA in GE group were significantly decreased （P<0.05，P<0.01）.The expression of CYP3A11 protein in GE group were significantly decreased （P<0.01）. Compared with the GE group， the GE + MP group displayed remarkably up-regulated BCRP protein and mRNA expression （P<0.05，P<0.01） and CYP3A11 protein expression （P<0.05）， but slightly up-regulated CYP3A11 mRNA expression. Compared with the GE group， the GE + Rif group exhibited down-regulated BCRP protein expression （P < 0.05）. The protein and mRNA expression levels of BCRP were lower in the GE + MP + Rif group than in the GE + MP group （P<0.05，P<0.01）. The PXR activator rifampicin regulated BCRP before and after the combination of G. elegans with M. pubescens. The CYP3A11 protein and mRNA expression levels in the GE + TCP group were higher than those in the GE group （P<0.05，P<0.01）. Compared with the GE + MP group， the GE + MP + TCP group showed up-regulated CYP3A11 protein and mRNA expression （P<0.05，P<0.01）. CAR activator TCPOBOP also had a regulatory effect on CYP3A11 before and after the compatibility of G. elegans with M. pubescens. ConclusionThe attenuated toxin after the combination of G. elegans with M. pubescens is closely related to the efflux transporter BCRP and the drug-metabolizing enzyme CYP3A11.
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Urea transporters (UT) play a vital role in the mechanism of urine concentration and are recognized as novel targets for the development of salt-sparing diuretics. Thus, UT inhibitors are promising for development as novel diuretics. In the present study, a novel UT inhibitor with a diarylamide scaffold was discovered by high-throughput screening. Optimization of the inhibitor led to the identification of a promising preclinical candidate,
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Aim: To investigate the effect of epithelial cell adhesion molecule (EpCAM) on metastasis and multidrug resistance of breast cancer and its mechanism. Methods Immunohistochemical staining was employed to detect the expression of EpCAM in adjacent non-tumor tissues (ANTTs) and breast cancer tissues. siRNA was applied to knock down EpCAM expression in MDA-MB-231 cells. Transwell assay was used to detect the invasion and migration ability of breast cancer cells. Western blot analysis was performed to determine the protein expression of EpCAM, epithelial mesenchymal transition (EMT) markers E-cadherin, N-cadherin and vimentin, breast cancer resistance protein (BCRP), and β-catenin. Results EpCAM immunoreactivity was consistently stronger in primary breast cancer tissues and even higher in metastatic lesions than that in ANTTs. The expression of EpCAM was significantly upregulated in triple negative breast cancer MDA-MB-231 cells. EpCAM knockdown using siRNA decreased the invasion and migration ability and BCRP expression, and partially reversed the EMT phenotypes of MDA-MB-231 cells, β-catenin expression was upregulated in MDA-MB-231 cells. ICG-001, a specific Wnt/β-catenin pathway inhibitor, downregulated the expression levels of EpCAM, N-cadherin, and vimentin in MDA-MB-231 cells. Conclusions EpCAM could promote metastasis and drug resistance of breast cancer through the induction of EMT, which is related to the Wnt/β-catenin signaling pathway.
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Objective:To compare the contents of alkaloids from fine and ultrafine powder of Dendrobium nobile stem in rat plasma,and investigate the effect of D. nobile stem with different particle sizes on gene expression of intestinal transporters. Method:Rats were randomly divided into the blank group,fine powder group of D. nobile stem(0.25 g·kg-1) and ultrafine powder group of D. nobile stem(0.25 g·kg-1).The rats were gavaged every 6 h for 5 days.The samples of rat plasma and small intestine were collected.The plasma samples were detected with UPLC-MS.The chromatography separation was performed on a Hypersil Gold C18 column(2.1 mm×150 mm,1.9 μm) with acetonitrile-0.1%formic acid solution as mobile phase for gradient elution.Electrospray ionization (ESI) was applied and operated in positive ion mode.The mRNA expression of multidrug resistance protein 1(MDR1),oligopeptide transporter protein 1(PEPT1),organic cation transporter protein 2(OCT2),breast cancer resistance protein 1(BCRP1),monocarboxylate transport protein 1(MCT1) and multidrug resistance related protein 2(MRP2) in small intestine were quantified by real time fluorescence quantitative polymerase chain reaction. Result:After intragastric administration of fine and ultrafine powder of D. nobile stem,dendrobine,mubironine B and dendramine could be detected in rat plasma.The contents of dendrobine and dendramine in the ultrafine powder group were significantly higher than that in the fine powder group(PD. nobile stem(PPD. nobile stem(PConclusion:Compared with the fine powder group of D. nobile stem,the plasma concentrations of dendrobine and dendramine in the ultrafine powder group are significantly increased,it may be related to the intestinal transporters of MDR1 and BCRP1.These results can provide experimental basis for selecting particle size of D. nobile stem.
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Aim To investigate the effect of baicalein on the reversal of multidrug resistance ( MDR) media-ted by breast cancer resistance protein ( BCRP) in hu-man breast cancer MCF-7/MX cells, and explore the possible mechanisms. Methods MTT assay was per-formed to determine the cytotoxicity of baicalein and susceptibility of chemotherapeutic drugs. The protein expression levels of BCRP, p-p38 MAPK and NF-κB p65 were determined by Western blot. Results MCF-7/MX cells were not only resistant to MX but cross-re-sistant to 5-FU and DDP, and the resistance index was 70. 45, 6. 68 and 21. 47, respectively. 2. 5, 5μmol· L-1 of baicalein could increase the sensitivity to above chemotherapeutic agents and decrease the expression levels of BCRP, p-p38 MAPK and NF-κB p65 in MCF-7/MX cells. Conclusion Baicalein can effec-tively reverse MDR of MCF-7/MX by down-regulating BCRP expression through p38/MAPK and NF-κB path-ways.
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ABSTRACT The present study describes the impact of chrysosplenetin, in the absence and presence of artemisinin, on in vitro breast cancer resistance protein-mediated transport activity in Caco-2 cell monolayers using aristolochic acid I as a specific probe substrate. We observed that novobiocin, a known breast cancer resistance protein active inhibitor, increased Papp (AP-BL) of aristolochic acid I 3.13 fold (p < 0.05) but had no effect on Papp (BL-AP). Efflux ratio (PBA/PAB) declined 4.44 fold (p < 0.05). Novobiocin, consequently, showed a direct facilitation on the uptake of AAI instead of its excretion. Oppositely, both artemisinin and chrysosplenetin alone at dose of 10 µM significantly decreased Papp (BL-AP) instead of Papp (AP-BL). Chrysosplenetin alone attenuated the efflux ratio, which was suggestive of being as a potential breast cancer resistance protein suppressant. Oddly, Papp (BL-AP) as well as efflux ratio were respectively enhanced 2.52 and 2.58 fold (p < 0.05), when co-used with artemisinin and chrysosplenetin in ratio of 1:2. The potential reason remains unclear; it might be relative to binding sites competition between artemisinin and chrysosplenetin or the homodimer/oligomer formation of breast cancer resistance protein bridged by disulfide bonds, leading to an altered in vitro breast cancer resistance protein-mediated efflux transport function.
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Objective To study the effects of different transport protein on the transport of 7,4'-dihydroxyflavone (7,4'-DHF) and its metabolite (7,4'-DHF-S) in Caco-2 cell model.Methods Ultra performance liquid chromatography was employed to determinethe content of 7,4'-DHF and 7,4'-DHF-S incubation buffer,their structures were identified by LC-MS/MS.Bidirectional transport of Caco-2 cells model was used to investigate the influence of ko143 (the inhibitor of BCRP) and MK571 (the inhibitor of MRP2) on the transport of 7,4'-DHF and 7,4'-DHF-S,respectively.Results Metabolic product of 7,4'-DHF in Caco-2 monolayer cell was identified as one monosulfate;PDR of 7,4'-DHF was (1.43 ± 0.11),PDR of ko143 and MK571 on the apparent permeability of 7,4'-DHF was (1.59 ± 0.04) and (1.48 ± 0.07) (P ＞ 0.05);PDR of 7,4'-DHF-S was (1.60 ± 0.06);ko143 could significantly reduce the apparent permeability of 7,4'-DHF-S,and the PDR was (0.23 ±0.03) (P ＜ 0.01);MK571 had no significant effect on the apparent permeability of the 7,4'-DHF-S,and the PDR was (1.51±0.04) (P ＞ 0.05).Conclusion Caco-2 cells can mediate the suffonated reaction of 7,4'-DHF;7,4'-dihydroxyflavone sulfonated combination product may be a substrate for BCRP.
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Human breast cancer resistance protein (BCRP), an ATP-binding cassette (ABC) efflux transporter, is mainly responsible for the transport of many endogenous and xenobiotics. BCRP is expressed in different tissues, such as placental, intestinal epithelium, endothelial cells of brain microvessels, and renal proximal tubular cells. BCRP is considered as one of the key factor for the drug-drug interaction and individual difference in drug therapy. The review will provide an overview of the current knowledge on the discovery of BCRP and its physical function, transport mechanism, substrate and inhibitors, as well as its effect on the drug pharmacokinetics.
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Tamoxifen (TAM) is the most common nonsteroidal antiestrogen agent, which has been widely used in the prevention of recurrence of estrogen or progesterone receptor-positive breast cancer in patients. It is metabolized by cytochrome P450 oxidases to its active metabolite (4-hydroxytamoxifen, 4-OH-TAM) and endoxifen (EDF), which played a critical role in the therapy. 4-OH-TAM and EDF have 30-to 100-fold more potency than TAM in the suppression of estrogen-dependent breast cancer cell proliferation. CYP3A4 and CYP2D6, as the key drug-metabolizing enzymes in those metabolic actions, are known to have several alleles. Genetic polymorphisms of CYP2D6 and CYP3A4 will influence the plasma concentrations of active TAM metabolites and clinical outcomes for breast cancer patients treated with TAM. The genetic polymorphisms of drug transporters, involved in the disposition of active TAM metabolites, also have the potential to influence the plasma concentrations of active TAM metabolites and clinical outcome for the treatment of breast cancer. In this review, we summarized the association of the genetic polymorphisms in the metabolic enzymes and transporters involved in the metabolism and disposition of TAM with the metabolite concentration, efficacy and adverse effects of TAM, which provides a fundamental reference for further pharmacogenomic study and clinical use of TAM.
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Objective To investigate influences of paeonol on mRNA expression and function of drug transporters BCRP and SLCO1B1 in HepG2 cell.Methods Cell counting Kit-8 assay was used to detect the viability of HepG2 cells;Real-time fluorescent quantitative PCR (qPCR) was performed to measure the expressions of BCRP and SLCO1B1 mRNAs; flow cytometry was applied to determine the transport functions of BCRP and SLCO1B1. Results Paeonol (2-8μg/mL) did not decrease HepG2 cell survival rate, but 16 μg/mL paeonol significantly reduced cell survival rate (P<0.05). Paeonol(2-8μg/mL)significantly induced the mRNA expression and function of drug transporters BCRP and SLCO1B1(P <0.05).Compared with control group, transcription level of paeonol group’s BCRP and SLCO1B1 drug transporters obviously up-regulated, the of translocation efficiency of substrate specificity increased significantly (P <0.05).Conclusion Paeonol can induce drug hepatocellular transporters BCRP and SLCO1B1 gene expression, thereby promote the substrate transport the transmembrane.It is indicated that the drug combination of paeonol and BCRP and SLCO1B1 transporters, there may be a risk of drug interactions.
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As an important ABC transporter, breast cancer re-sistance protein ( BCRP) plays an important role in tumor multi-drug resistance. Many laboratories are focusing on BCRP to re-verse multidrug resistance. We summarize in the paper the re-search progress on the regulation of BCRP expression, subcellu-lar localization, ATP-dependence, inhibition or modulation of its transport activity and potential clinical treatment strategies in or-der to provide theoretical support and some new research ideas for the reverse of multidrug resistance in clinic.
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Background and purpose:Resistance to antitumor agents is a major cause of treatment failure in patients with breast cancer. Research has shown that, tumor stem cell marker aldehyde dehydrogenase 1 (ALDH1) is related with some anticancer drugs (such as cyclophosphamide, cisplatin) resistance, and the content of ALDH1 in tumor cells after treatment is higher than that before treatment. Breast cancer resistance protein (BCRP) is not expressed in normal tissues, but high expressed in breast cancer of after treatment, it may be associated with the mechanism of tumor drug resistance. This study was to investigate the correlation between expression and the relationship between these two kinds of protein ALDH1, BCRP and clinical pathological characteristics. Methods:Immunohistochemistry was performed to detect the expression of ALDH1 and BCRP in breast inifltrating ductal carcinoma tissues, and whether there is a correlation between and explore their relationship with clinical pathological features and their expression. Results:The expression of ALDH1 protein and BCRP protein in breast cancer and paracarcinoma breast tissues has signiifcant difference(χ2=14.685, P=0.000;χ2=12.243, P=0.000).The expression of ALDH1 with patients age, pathologic stage, axillary lymph node metastasis, histological grading, ER, PR, and HER-2 state were not relevant(P>0.05). HER-2, BCRP protein, expression was higher in cancer tissue (χ2=5.289, P=0.021). There were no relevant with the expression of BCRP with patients age, pathologic stage, axillary lymph node metastasis and histological grading, estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor (HER-2) (P>0.05). Conclusion:ALDH1 proteins may be an independent factor compared with occur drug resistant protein, and participate breast cancer drug resistance in the chemotherapy and tumor invasion and metastasis of malignant biological behavior.
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Objective To investigate the correlation and significance of cyclooxygenase-2 (COX-2)and the breast cancer resistance protein (BCRP) expressions in non-Hodgkin lymphoma(NHL). Methods Ten patients with reactive lymph nodes (RLN) were considered as a control group. Compared with β-actin as internal control, the BCRP mRNA expressions of 61 NHL samples and 10 lymph tissues in control group were detected by semi-quantitative revere transcription polymerase chain reaction (RT-PCR) assay, while the expressions of COX-2 protein of the above specimens were detected by SP immunohistochemistry. Results The positive rates of COX-2 and BCRP in NHL were 50.8 %(31/61) and 45.8 % (28/61), respectively , and were higher than those in the control group (P ＜0.05). The expression of COX-2 was statistically positive correlated to that of BCRP (X2 =8.795, r=0.355, P＜0.05). The expressions of COX-2 and BCRP were not correlated to clinical and pathological factors, such as age, sex, IPI, LDH, β2-microglobulin level and Ann Arbor stage, however, the expression of BCRP was statistically correlated to chemotherapy efficacy.Conclusion BCRP may be involved in multi-drug resistance (MDR) of NHL, so it may contribute to the assessment of chemotherapy and prediction of NHL. Since there is a strong correlation between COX-2 expression and MDR in NHL, the application of COX-2 inhibitors may enhance sensitivity of chemotherapy.
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Objective: To establish a human multi-drug resistant embryonic kidney epithelial cell line HEK293/BCRP and investigate its biological characterization. Methods: The expression vector containing breast cancer resistance protein (BCRP) gene was constructed and transfected into HEK293 cells mediated by liposomes. The expression of BCRP was detected by RT-PCR, indirect immunofluorescence staining, and flow cytometry. The multidrug resistance index to mitoxantrone was measured by in vitro cytotoxicity test. Efflux of Rbodamine 123 (Rh123) and adriamycin (ADR) was determined by flow cytometry and laser scanning confocal microscopy, respectively. Results: BCRP expression significantly increased at both mRNA and protein levels in HEK293/BCRP cells. The resistance index to mitoxantrone was 112.07 times. The intracellular concentrations of Rb123 decreased by 42.25% and 69.01% after 1.5-h and 3-h efflux, respectively. Laser scanning confocal microscopy showed that the concentration of ADR also decreased. Conclusion: We successfully establish HEK293/BCRP cell line highly expressed BCRP and has cross-resistance to several anti-cancer drugs.
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Objective To observe the expression of breast cancer resistance protein(BCRP) gene in breast cancer patients,and to explore its value in evaluating the therapeutic effect of chemotherapy for breast cancer.Methods Semi-quantitative reverse-transcription polymerase chain reaction(RT-PCR) was applied for the detection of mammary BCRP gene expression level in 60 female breast cancer patients who have received chemical therapeutic regimen of CEF(cyclophosphamide,epirubicin and 5-fluorouracil).The patients were divided into two groups according to the BCRP gene expression before CEF therapy.After treatment,the therapeutic effect in the two groups was observed and the BCRP gene expression level was also measured.Results In 17(17/60,28.3%) patients with positive BCRP gene expression,11 were relieved,with a relief rate of 64.7%,lower than 93.0% in patients with negative BCRP gene expression(P
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Purpose: The aim of the present study was to investigate the expression of breast cancer resistance protein (BCRP) in primary breast carcinoma and to determine whether such expression can predict survival. Methods: Expression of BCRP in 60 breast cancer patients was determined by immunohistochemistry on formalin-fixed, paraffin-embedded tumor section. The relationship between the expression of BCRP with the clinicphathological characteristics and the prognosis of breast cancer patients were also analyzed. Results: ①BCRP expression was observed in 21 of 60 (35%) cases.② BCRP expression was more frequently observed in patients with lymph node metastasis or hormone receptor (P 0. 05) .③Kaplan-Meier analyses revealed that BCRP expression associated only with disease-free survival (DFS) (P 0. 05) . ④In univariate and multivariate Cox regression analyses, tumor size, lymph node metastasis and estrogen receptor (ER) was associated with DFS and OS (P
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Background and purpose:BCRP was recently discoveried as a membrane transport protein affiliated with multiple drug resistance(MDR).The aim of present study was to investigate the expression of breast cancer resistance protein(BCRP) in primary breast carcinoma and its potential significance of guiding breast carcinoma chemotherapy and to determine whether such expression can be used as a predict factor for chemosensitivity.Methods:Expression of BCRP in 31 primary breast carcinoma tissues was determined by flow cytometry.The relationship between the expression of BCRP with the clinicpathological characteristics and the prognosis of breast carcinoma patients was also analyzed.Results:The expression level of BCRP was higher in breast carcinoma tissue(0.282581?0.183686) than control group(0.03125?1.000905).There was no statistical difference befween BCRP and state of ER,PR,C-erbB_2 and EGFR in breast carcinoma tissues.In addition,it was the same situation no matter whether axillary lymph node was metastatasied or not.Conclusions:The results suggest that BCRP is expressed in primary breast carcinoma and is a cell membrane protein independent on ER,PR,c-erb-B-2,EGFR.Based chemotherapy is more effective for the patients with the overexpression of BCRP in primary breast carcinoma tissue.