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Steroid Resistance Associated with High MIF and P-gp Serum Levels in SLE Patients. Molecules 2022; 27:molecules27196741. [PMID: 36235275 PMCID: PMC9573564 DOI: 10.3390/molecules27196741] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/14/2022] [Revised: 10/07/2022] [Accepted: 10/08/2022] [Indexed: 11/16/2022] Open
Abstract
Approximately 30% of patients with systemic lupus erythematosus (SLE) present steroid resistance (SR). Macrophage migration inhibition factor (MIF) and P-glycoprotein (P-gp) could be related to SR. This work aims to evaluate the relationship between MIF and P-pg serum levels in SR in SLE. Methods: Case−control study including 188 SLE patients who were divided into two groups (90 in the steroid-resistant group and 98 in the steroid-sensitive (SS) group) and 35 healthy controls. MIF and P-gp serum levels were determined by ELISA. Multivariable logistic regression and chi-squared automatic interaction detection (CHAID) were used to explore risk factors for SR. Results: The steroid-resistant group presented higher MIF and P-gp serum levels in comparison with the SS (p < 0.001) and reference (p < 0.001) groups. MIF correlated positively with P-gp (rho = 0.41, p < 0.001). MIF (≥15.75 ng/mL) and P-gp (≥15.22 ng/mL) were a risk factor for SR (OR = 2.29, OR = 5.27). CHAID identified high P-gp as the main risk factor for SR and high MIF as the second risk factor in those patients with low P-gp. Conclusions: An association between MIF and P-gp serum levels was observed in SR. CHAID identified P-gp ≥ 15.22 ng/mL as the main risk factor for SR. More studies are needed to validate these results.
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Perez-Guerrero EE, Gonzalez-Lopez L, Muñoz-Valle JF, Vasquez-Jimenez JC, Ramirez-Villafaña M, Sanchez-Rodriguez EN, Gutierrez-Ureña SR, Cerpa-Cruz S, Aguilar-Chavez EA, Cardona-Muñoz EG, Vazquez-Villegas ML, Saldaña-Cruz AM, Rodriguez-Jimenez NA, Fajardo-Robledo NS, Gamez-Nava JI. Serum P-glycoprotein level: a potential biomarker of DMARD failure in patients with rheumatoid arthritis. Inflammopharmacology 2018; 26:10.1007/s10787-018-0529-2. [PMID: 30209762 DOI: 10.1007/s10787-018-0529-2] [Citation(s) in RCA: 6] [Impact Index Per Article: 0.9] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/24/2018] [Accepted: 09/01/2018] [Indexed: 10/28/2022]
Abstract
OBJECTIVES To evaluate the utility of elevated serum P-glycoprotein (P-gp) as a risk marker of therapeutic response failure in rheumatoid arthritis (RA) patients treated with disease-modifying antirheumatic drugs (DMARDs). METHODS A cross-sectional study was conducted in 151 RA patients. Patients were classified into two groups according to the response achieved in terms of the disease activity score (DAS)28 after ≥ 6 months: (1) patients with a therapeutic response to DMARDs, with DAS28 < 3.2; and (2) patients without a response to DMARDs, with persistent DAS28 ≥ 3.2. We explored a wide group of clinical factors associated with therapeutic resistance. Serum P-gp levels were measured by ELISA. The risk of P-gp elevation as a marker of failure to achieve a therapeutic response to DMARDs was computed using multivariate logistic regression. RESULTS Serum P-gp levels were significantly higher in RA patients (n = 151) than in the controls (n = 30) (158.70 ± 182.71 ng/mL vs. 14.12 ± 8.97 ng/mL, p < 0.001). The P-gp level was correlated with the DAS28 score (r = 0.39, p < 0.001). RA patients with DMARD failure had higher serum P-gp levels than patients with a therapeutic response (206 ± 21.47 ng/mL vs 120.60 ± 15.70 ng/mL; p = 0.001). High P-gp levels increased the risk of DMARD failure (OR 3.36, 95% CI 1.54-7.27, p = 0.001). After adjusting for confounding variables, elevated P-gp remained associated with DMARD failure (OR 2.64, 95% CI 1.29-5.40, p = 0.01). CONCLUSION Elevated serum P-gp is associated with DMARD failure. The P-gp level can be considered a clinical tool for evaluating the risk of DMARD failure in patients; however, future prospective studies should be performed to evaluate the utility of this marker in predicting long-term responses.
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Affiliation(s)
- E E Perez-Guerrero
- Instituto de Investigación en Ciencias Biomédicas, Centro Universitario de Ciencias de la Salud (CUCS), Universidad de Guadalajara (U de G), Guadalajara, Jalisco, Mexico
| | - L Gonzalez-Lopez
- Programa de Doctorado en Farmacología, CUCS, U de G, Guadalajara, Jalisco, Mexico
- Hospital General Regional 110, Instituto Mexicano del Seguro Social (IMSS), Guadalajara, Jalisco, Mexico
| | - J F Muñoz-Valle
- Instituto de Investigación en Ciencias Biomédicas, Centro Universitario de Ciencias de la Salud (CUCS), Universidad de Guadalajara (U de G), Guadalajara, Jalisco, Mexico
| | - J C Vasquez-Jimenez
- Centro Universitario de Investigaciones Biomédicas, Universidad de Colima, Colima, Mexico
| | - M Ramirez-Villafaña
- Programa de Doctorado en Ciencias Médicas, Universidad de Colima, Colima, Mexico
- Unidad de Investigación Biomédica 02, UMAE, Hospital de Especialidades, Centro Médico Nacional de Occidente, IMSS, Guadalajara, Jalisco, Mexico
| | | | - S R Gutierrez-Ureña
- División de Reumatología, Hospital Civil de Guadalajara "Fray Antonio Alcalde", Guadalajara, Jalisco, México
| | - S Cerpa-Cruz
- División de Reumatología, Hospital Civil de Guadalajara "Fray Antonio Alcalde", Guadalajara, Jalisco, México
| | - E A Aguilar-Chavez
- Unidad de Medicina Familiar 2, IMSS, Guadalajara, Jalisco, Mexico
- Centro Universitario de Tonalá, U de G, Tonalá, Jalisco, Mexico
| | | | - M L Vazquez-Villegas
- Departamento de Epidemiología, Unidad Médica Familiar 4, IMSS, Guadalajara, Jalisco, Mexico
- Departamento de Salud Pública, CUCS, U de G, Guadalajara, Jalisco, Mexico
| | | | | | - N S Fajardo-Robledo
- Laboratorio de Investigación y Desarrollo Farmacéutico, Centro Universitario de Ciencias Exactas e Ingenierías, U de G, Guadalajara, Jalisco, Mexico
| | - J I Gamez-Nava
- Programa de Doctorado en Farmacología, CUCS, U de G, Guadalajara, Jalisco, Mexico.
- Unidad de Investigación Biomédica 02, UMAE, Hospital de Especialidades, Centro Médico Nacional de Occidente, IMSS, Guadalajara, Jalisco, Mexico.
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Wang CW, Huang CC, Chou PH, Chang YP, Wei S, Guengerich FP, Chou YC, Wang SF, Lai PS, Souček P, Ueng YF. 7-ketocholesterol and 27-hydroxycholesterol decreased doxorubicin sensitivity in breast cancer cells: estrogenic activity and mTOR pathway. Oncotarget 2017; 8:66033-66050. [PMID: 29029490 PMCID: PMC5630390 DOI: 10.18632/oncotarget.19789] [Citation(s) in RCA: 14] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/13/2017] [Accepted: 06/27/2017] [Indexed: 11/30/2022] Open
Abstract
Hypercholesterolemia is one of the risk factors for poor outcome in breast cancer therapy. To elucidate the influence of the main circulating oxysterols, cholesterol oxidation products, on the cell-killing effect of doxorubicin, cells were exposed to oxysterols at a subtoxic concentration. When cells were exposed to oxysterols in fetal bovine serum-supplemented medium, 7-ketocholesterol (7-KC), but not 27-hydroxycholesterol (27-HC), decreased the cytotoxicity of doxorubicin in MCF-7 (high estrogen receptor (ER)α/ERβ ratio) cells and the decreased cytotoxicity was restored by the P-glycoprotein inhibitor verapamil. 7-KC stimulated the efflux function of P-glycoprotein and reduced intracellular doxorubicin accumulation in MCF-7 but not in ERα(-) MDA-MB-231 and the resistant MCF-7/ADR cells. In MCF-7 cells, 7-KC increased the mRNA and protein levels of P-glycoprotein. The 7-KC-suppressed doxorubicin accumulation was restored by the fluvestrant and ERα knockdown. In a yeast reporter assay, the ERα activation by 7-KC was more potent than 27-HC. 7-KC, but not 27-HC, stimulated the expression of an ER target, Trefoil factor 1 in MCF-7 cells. When charcoal-stripped fetal bovine serum was used, both 7-KC and 27-HC induced Trefoil factor 1 expression and reduced doxorubicin accumulation in MCF-7 cells. 7-KC-reduced doxorubicin accumulation could be reversed by inhibitors of phosphatidylinositol 3-kinase, Akt, and mammalian target of rapamycin (mTOR). These findings demonstrate that 7-KC decreases the cytotoxicity of doxorubicin through the up-regulation of P-glycoprotein in an ERα- and mTOR-dependent pathway. The 7-KC- and 27-HC-elicited estrogenic effects are crucial in the P-glycoprotein induction in breast cancer cells.
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Affiliation(s)
- Chun-Wei Wang
- National Research Institute of Chinese Medicine, Ministry of Health and Welfare, Taipei, Taiwan, R.O.C.,Institute of Biopharmaceutical Sciences, School of Pharmacy, National Yang-Ming University, Taipei, Taiwan, R.O.C
| | - Chiung-Chiao Huang
- National Research Institute of Chinese Medicine, Ministry of Health and Welfare, Taipei, Taiwan, R.O.C
| | - Pei-Hsin Chou
- Department of Environmental Engineering, National Chung-Kung University, Tainan, Taiwan, R.O.C
| | - Yu-Ping Chang
- National Research Institute of Chinese Medicine, Ministry of Health and Welfare, Taipei, Taiwan, R.O.C
| | - Shouzuo Wei
- Department of Medicine, Vanderbilt University School of Medicine, Nashville, TN, USA
| | | | - Yueh-Ching Chou
- Department of Pharmacology, School of Medicine, National Yang-Ming University, Taipei, Taiwan, R.O.C.,Department of Pharmacy, Taipei Veterans General Hospital, Taipei, Taiwan, R.O.C.,Department of Pharmacy, Taipei Medical University, Taipei, Taiwan, R.O.C
| | - Sheng-Fan Wang
- National Research Institute of Chinese Medicine, Ministry of Health and Welfare, Taipei, Taiwan, R.O.C.,Department of Pharmacy, Taipei Veterans General Hospital, Taipei, Taiwan, R.O.C
| | - Ping-Shan Lai
- Department of Chemistry, College of Science, National Chung-Hsin University, Taichung, Taiwan, R.O.C
| | - Pavel Souček
- Department of Toxicogenomics, National Institute of Public Health, Prague, Czech Republic
| | - Yune-Fang Ueng
- National Research Institute of Chinese Medicine, Ministry of Health and Welfare, Taipei, Taiwan, R.O.C.,Institute of Biopharmaceutical Sciences, School of Pharmacy, National Yang-Ming University, Taipei, Taiwan, R.O.C.,Department of Pharmacology, School of Medicine, National Yang-Ming University, Taipei, Taiwan, R.O.C.,Institute of Medical Sciences, Taipei Medical University, Taipei, Taiwan, R.O.C
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Nocera AL, Meurer AT, Singleton A, Simons C, BuSaba J, Tara Gass N, Han X, Bleier BS. Intact soluble P-glycoprotein is secreted by sinonasal epithelial cells. Am J Rhinol Allergy 2017; 30:246-9. [PMID: 27456593 DOI: 10.2500/ajra.2016.30.4330] [Citation(s) in RCA: 6] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/20/2022]
Abstract
BACKGROUND P-glycoprotein (P-gp) is a 170 kDa transmembrane efflux pump, which is upregulated in chronic rhinosinusitis. Studies of leukemia demonstrated that P-gp may also be secreted in an intact soluble form. The purpose of this study was to explore whether sinonasal epithelial cells were capable of secreting soluble P-gp and whether P-gp has any functional role. METHODS Soluble and cytoplasmic P-gp were quantified in vehicle and lipopolysaccharide exposed cultures by enzyme-linked immunosorbent assay. The molecular weight of the soluble P-gp was determined by Western blot. Naive cultures were exposed to recombinant human P-gp at 0-2000 ng/mL. The degree of membranous interpolation was determined by quantitative fluorescent immunocytochemistry and function was determined by a calcein acetoxymethyl ester assay. RESULTS Soluble P-gp was secreted intact at 170 kDa. Mean (standard deviation) secretion was detected within vehicle wells at 55.43 ± 26.26 ng/mL, which significantly increased to 333.27 ± 305.98 ng/mL (p < 0.001) after lipopolysaccharide stimulation. Soluble P-gp strongly and significantly correlated with cytoplasmic P-gp (r = 0.57, p = 0.000001). Exposure to 2000 ng/mL of recombinant P-gp significantly increased corrected total cell fluorescence (1.34 ± 1.85) relative to vehicle control 0.29 ± 0.26 (p = 0.01) and significantly reduced calcein acetoxymethyl ester fluorescence (82.03 ± 43.69) relative to 100 ng/mL recombinant P-gp exposed cells (123.11 ± 42.16, p = 0.001). CONCLUSION Cultured sinonasal epithelial cells were able to both secrete intact P-gp and could functionally interpolate soluble P-gp into their cell membrane. These in vitro findings indicated that soluble P-gp may be present in nasal mucus as a biomarker and could participate in the maintenance of P-gp overexpression in chronic rhinosinusitis and associated inflammation.
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Affiliation(s)
- Angela L Nocera
- Massachusetts Eye and Ear Infirmary, Harvard Medical School, Boston, Massachusetts, USA
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Perez-Guerrero EE, Gamez-Nava JI, Muñoz-Valle JF, Cardona-Muñoz EG, Bonilla-Lara D, Fajardo-Robledo NS, Nava-Zavala AH, Garcia-Cobian TA, Rincón-Sánchez AR, Murillo-Vazquez JD, Cardona-Müller D, Vazquez-Villegas ML, Totsuka-Sutto SE, Gonzalez-Lopez L. Serum levels of P-glycoprotein and persistence of disease activity despite treatment in patients with systemic lupus erythematosus. Clin Exp Med 2017; 18:109-117. [PMID: 28243944 DOI: 10.1007/s10238-017-0459-0] [Citation(s) in RCA: 14] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/10/2016] [Accepted: 02/14/2017] [Indexed: 01/16/2023]
Abstract
Around 25% of patients with systemic lupus erythematosus (SLE) could be refractory to conventional therapies. P-glycoprotein expression on cell surface has been implied on drug resistance, however, to date, it is unknown if P-gp serum levels are associated with SLE disease activity. Evaluate the association of serum P-gp levels and SLE with disease activity despite treatment. A cross-sectional study was conducted on 93 female SLE patients, all receiving glucocorticoids at stable doses for the previous 6 months before to baseline. SLE patients were classified into two groups: (a) patients with active disease [SLE disease activity index (SLEDAI) ≥ 3] despite treatment, and (b) patients with inactive disease (SLEDAI < 3) after treatment. Forty-three healthy females comprised the control group. Serum P-gp, anti-DNA, and both anti-nucleosome antibody levels were measured using ELISA. Active-SLE patients despite treatment had higher P-gp levels compared with inactive-SLE after treatment (78.02 ng/mL ± 114.11 vs. 33.75 ng/mL ± 41.11; p = 0.018) or versus reference group subjects (30.56 ng/mL ± 28.92; p = 0.011). P-gp levels correlated with the scores of SLEDAI (r = 0.26; p = 0.01), Mexican-SLEDAI (MEX-SLEDAI) (r = 0.32; p = 0.002), SLICC/ACR damage index (r = 0.47; p < 0.001), and with prednisone doses (r = 0.33; p = 0.001). In the multivariate model, the high P-gp levels were associated with SLICC/ACR score (p = 0.001), and SLEDAI score (p = 0.014). Our findings support a relationship between serum P-gp levels and SLE with disease activity despite treatment, but it requires further validation in longitudinal studies.
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Affiliation(s)
- Edsaul Emilio Perez-Guerrero
- Programa de Doctorado en Farmacologia, Centro Universitario de Ciencias de la Salud (CUCS), Universidad de Guadalajara, Sierra Mojada 950, Col. Independencia, 44340, Guadalajara, Jalisco, Mexico
| | - Jorge Ivan Gamez-Nava
- Programa de Doctorado en Farmacologia, Centro Universitario de Ciencias de la Salud (CUCS), Universidad de Guadalajara, Sierra Mojada 950, Col. Independencia, 44340, Guadalajara, Jalisco, Mexico.,Unidad de Investigación Biomédica 02 (UIEC), UMAE Hospital de Especialidades, Centro Médico Nacional de Occidente, Instituto Mexicano del Seguro Social, Avenida Belisario Domínguez 1000, Col. Independencia Oriente, 44340, Guadalajara, Jalisco, Mexico
| | - Jose Francisco Muñoz-Valle
- Instituto de Investigación en Ciencias Biomédicas, Centro Universitario de Ciencias de la Salud (CUCS), Universidad de Guadalajara, Sierra Mojada 950, Col. Independencia, 44340, Guadalajara, Jalisco, Mexico
| | - Ernesto German Cardona-Muñoz
- Departamento de Fisiología, Centro Universitario de Ciencias de la Salud (CUCS), Universidad de Guadalajara, Sierra Mojada 950, Col. Independencia, 44340, Guadalajara, Jalisco, Mexico
| | - David Bonilla-Lara
- Programa de Doctorado en Farmacologia, Centro Universitario de Ciencias de la Salud (CUCS), Universidad de Guadalajara, Sierra Mojada 950, Col. Independencia, 44340, Guadalajara, Jalisco, Mexico
| | - Nicte Selene Fajardo-Robledo
- Laboratorio de Investigación y Desarrollo Farmacéutico, Centro Universitario de Ciencias Exactas e Ingenierías, Universidad de Guadalajara, Blvd. Marcelino García Barragán 421, 44430, Guadalajara, Jalisco, Mexico
| | - Arnulfo Hernan Nava-Zavala
- Unidad de Investigación Biomédica 02 (UIEC), UMAE Hospital de Especialidades, Centro Médico Nacional de Occidente, Instituto Mexicano del Seguro Social, Avenida Belisario Domínguez 1000, Col. Independencia Oriente, 44340, Guadalajara, Jalisco, Mexico.,Departamento de Inmunología y Reumatología, Hospital General de Occidente, Secretaria de Salud, Av Zoquipan 1050, Seattle, 45170, Zapopan, Jalisco, Mexico.,Programa Internacional de Medicina, Universidad de Autónoma de Guadalajara, Av. Patria 1201, Col. Lomas del Valle, 45129, Zapopan, Jalisco, Mexico
| | - Teresa Arcelia Garcia-Cobian
- Departamento de Fisiología, Centro Universitario de Ciencias de la Salud (CUCS), Universidad de Guadalajara, Sierra Mojada 950, Col. Independencia, 44340, Guadalajara, Jalisco, Mexico
| | - Ana Rosa Rincón-Sánchez
- Programa de Doctorado en Farmacologia, Centro Universitario de Ciencias de la Salud (CUCS), Universidad de Guadalajara, Sierra Mojada 950, Col. Independencia, 44340, Guadalajara, Jalisco, Mexico
| | - Jessica Daniela Murillo-Vazquez
- Programa de Doctorado en Farmacologia, Centro Universitario de Ciencias de la Salud (CUCS), Universidad de Guadalajara, Sierra Mojada 950, Col. Independencia, 44340, Guadalajara, Jalisco, Mexico
| | - David Cardona-Müller
- Departamento de Fisiología, Centro Universitario de Ciencias de la Salud (CUCS), Universidad de Guadalajara, Sierra Mojada 950, Col. Independencia, 44340, Guadalajara, Jalisco, Mexico
| | - Maria Luisa Vazquez-Villegas
- Programa de Doctorado en Farmacologia, Centro Universitario de Ciencias de la Salud (CUCS), Universidad de Guadalajara, Sierra Mojada 950, Col. Independencia, 44340, Guadalajara, Jalisco, Mexico.,Unidad Médica Familiar 4 y 8, Departamento de Epidemiologia, Instituto Mexicano del Seguro Social (IMSS), Fidel Velázquez Sánchez 1531, Atemajac del Valle, 44218, Guadalajara, Jalisco, Mexico
| | - Sylvia Elena Totsuka-Sutto
- Departamento de Fisiología, Centro Universitario de Ciencias de la Salud (CUCS), Universidad de Guadalajara, Sierra Mojada 950, Col. Independencia, 44340, Guadalajara, Jalisco, Mexico
| | - Laura Gonzalez-Lopez
- Programa de Doctorado en Farmacologia, Centro Universitario de Ciencias de la Salud (CUCS), Universidad de Guadalajara, Sierra Mojada 950, Col. Independencia, 44340, Guadalajara, Jalisco, Mexico. .,Departamento de Medicina Interna-Reumatología, Instituto Mexicano del Seguro Social (IMSS), Hospital General Regional 110, Av Circunvalación Oblatos 2208, Colonia Circunvalación Oblatos, 44716, Guadalajara, Jalisco, Mexico. .,, Avenida Salto del Agua 2192, Colonia Jardines del Country, 44210, Guadalajara, Mexico.
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Nocera AL, Meurer AT, Miyake MM, Sadow PM, Han X, Bleier BS. Secreted P-glycoprotein is a noninvasive biomarker of chronic rhinosinusitis. Laryngoscope 2016; 127:E1-E4. [PMID: 27577924 DOI: 10.1002/lary.26249] [Citation(s) in RCA: 19] [Impact Index Per Article: 2.1] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/07/2016] [Revised: 07/18/2016] [Accepted: 07/20/2016] [Indexed: 12/26/2022]
Abstract
OBJECTIVE The discovery of noninvasive biomarkers of chronic rhinosinusitis (CRS) is critical to enable our ability to provide prognostic information and targeted medical therapy. Epithelial P-glycoprotein (P-gp) is overexpressed in CRS and exists in an extracellular, secreted form. The objective of this study was to determine whether secreted P-gp concentrations are elevated in CRS and can be used to predict disease severity. METHODS Institutional review board-approved study examining mucus concentrations of P-gp in 36 patients (10 control, 16 CRS without nasal polyps [CRSsNP], and 10 CRS with nasal polyps [CRSwNP]). P-gp concentrations were determined by enzyme-linked immunosorbent assay and normalized to total protein (TP). Clinical indices of disease severity, including the Sino-Nasal Outcomes Test (SNOT-22) and Lund-Mackay score, were collected for all patients. RESULTS Secreted P-gp concentration was significantly higher in CRS versus control patients (mean ± standard deviation; 247.8 ± 224.8 vs. 102.4 ± 81.7 pcg P-gp/μg TP, P = 0.022). A threshold value of 250 pcg/μg TP was used to differentiate low versus high secretors. High P-gp secretors with CRS (sNP and wNP, n = 9) demonstrated significantly higher SNOT-22 and Lund-Mackay scores (57.1 ± 7.9 and 13.9 ± 7.3) versus low secretors (38.3 ± 23.9 and 6.8 ± 7.3; P = 0.030 and P = 0.013, respectively) and had a significantly higher proportion of CRSwNP (66.7%) versus the low secretors (23.5%, n = 17, P = 0.046). CONCLUSION P-gp secretion levels are significantly elevated in patients with CRS. High P-gp secretion is associated with a higher incidence of CRSwNP and confers worse subjective and objective measures of disease severity. The presence of elevated P-gp secretion may therefore represent a novel noninvasive biomarker of CRS and could be used to predict patients who may benefit from P-gp inhibitory therapeutic strategies. LEVEL OF EVIDENCE NA Laryngoscope, 127:E1-E4, 2017.
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Affiliation(s)
- Angela L Nocera
- Massachusetts Eye and Ear Infirmary, Harvard Medical School, Boston, Massachusetts, U.S.A
| | - Ana T Meurer
- Massachusetts Eye and Ear Infirmary, Harvard Medical School, Boston, Massachusetts, U.S.A
| | - Marcel M Miyake
- Massachusetts Eye and Ear Infirmary, Harvard Medical School, Boston, Massachusetts, U.S.A
| | - Peter M Sadow
- Department of Pathology, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts, U.S.A
| | - Xue Han
- Department of Biomedical Engineering, Boston University, Boston, Massachusetts, U.S.A
| | - Benjamin S Bleier
- Massachusetts Eye and Ear Infirmary, Harvard Medical School, Boston, Massachusetts, U.S.A
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Limtrakul P, Khantamat O, Pintha K. Inhibition of P-Glycoprotein Function and Expression by Kaempferol and Quercetin. J Chemother 2013; 17:86-95. [PMID: 15828450 DOI: 10.1179/joc.2005.17.1.86] [Citation(s) in RCA: 131] [Impact Index Per Article: 10.9] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/31/2022]
Abstract
The 170 kDa plasma membrane P-glycoprotein (Pgp) causes the efflux of chemotherapeutic drugs from cells and is believed to be an important mechanism in multidrug resistance (MDR) in human cancer. This study demonstrates that some putative flavonoids, i.e., flavonols (quercetin and kaempferol) and isoflavones (genistein and daidzein) markedly increase the sensitivity of the multidrug-resistant human cervical carcinoma KB-V1 cells (high Pgp expression) to vinblastine and paclitaxel dose-dependently, and also decrease the relative resistance of these anti-cancer-drugs in KB-V1 cells. None of the flavonoids had a significant effect on vinblastine and paclitaxel cytotoxicity in wildtype drug-sensitive KB-3-1 cells (lacking Pgp). These flavonoids also caused an increase in intracellular accumulation, and reduced the efflux of Rh123 and 3[H]vinblastine in KB-V1 cells, but not in KB-3-1 cells. The flavonols increased the inhibitory effectiveness of Pgp activity in MDR KB-V1 cells more than isoflavones. Only treatment with flavonols up to 48 h was able to significantly decrease the Pgp expression in a dose-dependent manner in KB-V1 cells. These findings provide evidence that flavonols reduced Pgp expression and function resulting in the inhibition of Pgp activity, but isoflavones modulated intracellular drug levels by inhibiting Pgp function with no effect on Pgp expression. Among the flavonoids tested, flavonols, particularly kaempferol, exhibit the most potent MDR reversing property in KB-V1 cells.
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Affiliation(s)
- P Limtrakul
- Department of Biochemistry, Faculty of Medicine, Chiang Mai University, Chiang Mai, Thailand.
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Chiampanichayakul S, Anuchapreeda S, Chruewkamlow N, Mahasongkram K, Thanaratanakorn P, Kasinrerk W. Production of monoclonal antibodies to P-glycoprotein: its application in detection of soluble and surface P-glycoprotein of leukemia patients. Int J Hematol 2010; 92:326-33. [DOI: 10.1007/s12185-010-0668-8] [Citation(s) in RCA: 10] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/19/2010] [Revised: 08/08/2010] [Accepted: 08/12/2010] [Indexed: 11/27/2022]
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Inhibition of P-glycoprotein-mediated docetaxel efflux sensitizes ovarian cancer cells to concomitant docetaxel and SN-38 exposure. Anticancer Drugs 2009; 20:267-76. [PMID: 19262372 DOI: 10.1097/cad.0b013e328329977f] [Citation(s) in RCA: 13] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/25/2022]
Abstract
The first-line treatment of ovarian cancer is based on cytoreductive surgery and the use of anticancer drugs. The main disadvantage in the usage of anticancer drugs is the wide capacity of cancer cells to acquire a resistance to chemotherapeutic agents and therefore new treatment strategies have to be developed and tested. In this study, the responses of seven ovarian carcinoma cell lines to docetaxel and a camptothecin derivative, SN-38, were evaluated. We further studied the expression of P-glycoprotein (P-gp), the best described mechanism of drug resistance, in these cells and the effect of treatment with a specific P-gp inhibitor (PGP-4008). Simultaneous treatment with docetaxel and SN-38 (docetaxel+SN-38) had an antagonistic growth effect that was not dependent on the administration schedule. Both drugs alone or in combination induced G2M cell cycle arrest. Docetaxel was a more potent inducer of apoptosis than SN-38, but simultaneous treatment with docetaxel+SN-38 decreased the proportion of apoptotic cells to the same level observed after exposure to SN-38 alone. SN-38 increased P-gp expression in all cell lines. PGP-4008 enhanced docetaxel-mediated growth inhibition and apoptosis, but it did not have an effect when used simultaneously with SN-38. When cells were treated with docetaxel, SN-38, and PGP-4008 simultaneously, the growth was inhibited more efficiently and the proportion of apoptotic cells was higher than that without PGP-4008. Thus, treatment of ovarian cancer cells with docetaxel+SN-38 may have antagonistic effects. The simultaneous administration of a P-gp inhibitor may prevent docetaxel efflux, thereby sensitizing cells to docetaxel and other chemotherapeutic agents.
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Zhang YS, Yuan FJ, Jia GF, Zhang JF, Hu LY, Huang L, Wang J, Dai ZQ. CIK cells from patients with HCC possess strong cytotoxicity to multidrug-resistant cell line Bel-7402/R. World J Gastroenterol 2005; 11:3339-45. [PMID: 15948236 PMCID: PMC4315985 DOI: 10.3748/wjg.v11.i22.3339] [Citation(s) in RCA: 38] [Impact Index Per Article: 1.9] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 02/06/2023] Open
Abstract
AIM: To investigate the cytotoxicity of the cytokine-induced killer (CIK) cells from the post-operation patients with primary hepatocellular carcinoma (HCC) to multidrug-resistant (MDR) cell of HCC both in vitro and in vivo.
METHODS: A drug-resistant cell line was established by culturing human HCC cell line Bel-7402 in complete RPMI 1640 medium with increasing concentrations of adriamycin from 10 to 2000 nmol/L. CIK cells were obtained by inducing the peripheral blood mononuclear cells with rhIFN-γ, monoclonal anti-CD3 antibody, rhIL-1α as well as rhIL-2, which were added into the culture. To detect the cytotoxicity of the CIK cells from HCC patients, the Bel-7402/R was taken as target (T) cells and CIK cells as effect (E) cells. Cytotoxic test was performed and measured by MTT. As to in vivo test, CIK cells were transfused into patients with HCC. The tumor specimens of the patients were obtained and immunohistochemistry was carried out to detect CD3, CD45, CD45RO as well as CD68.
RESULTS: A MDR 1 HCC cell line Bel-7402/R was established. Its MDR1 mRNA overexpressed which was shown by RT-PCR; the P-glycoprotein expression increased from 1.32% of parent cells to 54%. CIK cells expanded vigorously by more than 70-fold and the CD3+CD56+ increased by more than 600-fold after 3-wk incubation on average. The cytotoxicity of CIK from HCC patients to Bel-7402/R was about 50% and to L-02 below 10% (t = 8.87, P<0.01), the same as that of CIK from normal individuals. Each of the 17 patients received 1-5×1010 of CIK cell transfusion. No side effects were observed. After CIK treatment, the tumor tissue nodules formed and a large amount of lymphocytes infiltrated in the liver cancer tissue and CD3, CD45, CD45RO, and CD68 increased greatly which was shown by immunohistochemistry.
CONCLUSION: A stable MDR1 HCC cell line has been established which could recover from liquid nitrogen and CIK from HCC patients has strong cytotoxicity to MDR HCC cell. CIK adoptive immunotherapy is safe and has no side effects. Receivers improved their immunity to tumor evidently. CIK treatment may be a better choice for HCC patients after operation to prevent the recurrence, especially when tumors have developed drug resistance.
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Affiliation(s)
- You-Shun Zhang
- Institute of Liver Surgery, Dongfeng Hospital of YunYang Medical College, Shiyan 442008, Hubei Province, China
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11
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Novotna M, Libra A, Kopecky M, Pavek P, Fendrich Z, Semecky V, Staud F. P-glycoprotein expression and distribution in the rat placenta during pregnancy. Reprod Toxicol 2004; 18:785-92. [PMID: 15279876 DOI: 10.1016/j.reprotox.2004.04.014] [Citation(s) in RCA: 56] [Impact Index Per Article: 2.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/25/2004] [Revised: 04/16/2004] [Accepted: 04/27/2004] [Indexed: 11/20/2022]
Abstract
P-glycoprotein (P-gp) is a drug efflux transporter that limits the entry of various potentially toxic drugs and xenobiotics into the fetus and is thus considered a placental protective mechanism. In this study, P-gp expression was investigated in the rat chorioallantoic placenta over the course of pregnancy. Three methods have been employed: real-time RT-PCR, western blotting and immunohistochemistry. The expression of mdr1a and mdr1b genes was demonstrated as early as on the 11th gestation day (gd) and increased with advancing gestation. Western blotting analysis revealed the presence of P-gp in the rat placenta starting from gd 13 onwards. P-gp was localized in the developing labyrinth zone of the placenta on gd 13; from gd 15 up to the term P-gp was seen as a dot like continuous line in the syncytiotrophoblast layers. Our data confirm the presence of P-gp in the rat chorioallantoic placenta starting soon after its development, which may signify the involvement of P-gp in transplacental pharmacokinetics during the whole period of placental maturing.
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Affiliation(s)
- Martina Novotna
- Department of Pharmacology and Toxicology, Faculty of Pharmacy in Hradec Králové, Charles University in Prague, Czech Republic
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12
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Limtrakul P, Khantamat O, Pintha K. Inhibition of P-glycoprotein activity and reversal of cancer multidrug resistance by Momordica charantia extract. Cancer Chemother Pharmacol 2004; 54:525-30. [PMID: 15248030 DOI: 10.1007/s00280-004-0848-4] [Citation(s) in RCA: 52] [Impact Index Per Article: 2.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/19/2004] [Accepted: 04/21/2004] [Indexed: 11/27/2022]
Abstract
PURPOSE Multidrug resistance (MDR) is known as a problem limiting the success of therapy in patients treated long term with chemotherapeutic drugs. The drug resistance is mainly due to the overexpression of the 170 kDa P-glycoprotein (Pgp), which causes a reduction in drug accumulation in the cancer cells. In this study, novel chemical modulator(s) from bitter melon (Momordica charantia L.) extracts obtained from leaves, fruits and tendrils were tested for their abilities to modulate the function of Pgp and the MDR phenotype in the multidrug-resistant human cervical carcinoma KB-V1 cells (high Pgp expression) in comparison with wildtype drug-sensitive KB-3-1 cells (lacking Pgp). METHODS The KB-V1 and KB-3-1 cells were exposed to bitter melon extracts in the presence of various concentrations of vinblastine, and cytotoxicity was assessed by means of the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) colorimetric assay. Relative resistance was calculated as the ratio of the IC50 value of the KB-V1 cells to the IC50 value of the KB-3-1 cells. Accumulation and efflux of vinblastine in KB-V1 and KB-3-1 cells were measured using a [3H]-vinblastine incorporation assay. RESULTS The leaf extracts increased the intracellular accumulation of [3H]-vinblastine in KB-V1 cells in a dose-dependent manner, but extracts from the fruits and tendrils had no effect. By modulating Pgp-mediated vinblastine efflux, the leaf extracts decreased the [3H]-vinblastine efflux in KB-V1 cells in a dose-dependent manner, but not in KB-3-1 cells. Treatment of drug-resistant KB-V1 cells with bitter melon leaf extracts increased their sensitivity to vinblastine, but similar treatment of KB-3-1 cells showed no modulating effect. The fruit and tendril extracts did not affect the MDR phenotype in either cell line. CONCLUSION The leaf extracts from bitter melon were able to reverse the MDR phenotype, which is consistent with an increase in intracellular accumulation of the drug. The exact nature of the active components of bitter melon leaf extracts remains to be identified.
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Affiliation(s)
- Pornngarm Limtrakul
- Department of Biochemistry, Faculty of Medicine, Chiang Mai University, Chiang Mai, Thailand.
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13
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Jodoin J, Demeule M, Fenart L, Cecchelli R, Farmer S, Linton KJ, Higgins CF, Béliveau R. P-glycoprotein in blood-brain barrier endothelial cells: interaction and oligomerization with caveolins. J Neurochem 2004; 87:1010-23. [PMID: 14622130 DOI: 10.1046/j.1471-4159.2003.02081.x] [Citation(s) in RCA: 85] [Impact Index Per Article: 4.0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/12/2023]
Abstract
P-glycoprotein (P-gp), an adenosine triphosphate (ATP)-binding cassette transporter which acts as a drug efflux pump, is highly expressed at the blood-brain barrier (BBB) where it plays an important role in brain protection. Recently, P-gp has been reported to be located in the caveolae of multidrug-resistant cells. In this study, we investigated the localization and the activity of P-gp in the caveolae of endothelial cells of the BBB. We used an in vitro model of the BBB which is formed by co-culture of bovine brain capillary endothelial cells (BBCEC) with astrocytes. Caveolar microdomains isolated from BBCEC are enriched in P-gp, cholesterol, caveolin-1, and caveolin-2. Moreover, P-gp interacts with caveolin-1 and caveolin-2; together, they form a high molecular mass complex. P-gp in isolated caveolae is able to bind its substrates, and the caveolae-disrupting agents filipin III and nystatin decrease P-gp transport activity. In addition, mutations in the caveolin-binding motif present in P-gp reduced the interaction of P-gp with caveolin-1 and increased the transport activity of P-gp. Thus, P-gp expressed at the BBB is mainly localized in caveolae and its activity may be modulated by interaction with caveolin-1.
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Affiliation(s)
- Julie Jodoin
- Laboratoire de médecine moléculaire, Centre de cancérologie Charles Bruneau, Université du Québec á Montréal, Hôpital Sainte-Justine, Montréal, Québec, Canada
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14
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Nagashige M, Ushigome F, Koyabu N, Hirata K, Kawabuchi M, Hirakawa T, Satoh S, Tsukimori K, Nakano H, Uchiumi T, Kuwano M, Ohtani H, Sawada Y. Basal Membrane Localization of MRP1 in Human Placental Trophoblast. Placenta 2003; 24:951-8. [PMID: 14580377 DOI: 10.1016/s0143-4004(03)00170-x] [Citation(s) in RCA: 76] [Impact Index Per Article: 3.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 10/27/2022]
Abstract
The placental trophoblast is considered to act as a barrier between mother and fetus, mediating the exchange of various materials across the placenta. ATP-binding cassette (ABC) transporters such as P-glycoprotein (P-gp) and multidrug-resistance protein (MRP) are expressed in the placenta and function as efflux transport systems for xenobiotics. In the present study, we aimed to determine the localization of MRP1 in the human placenta in comparison with that of P-gp. Western blotting analysis with human placental membrane vesicles indicated that P-gp and MRP1 are localized on the brush-border membranes and basal membranes, respectively. Immunohistochemical analysis with human normal full-term placenta showed that anti-P-gp monoclonal antibody F4 stained the brush-border side of the trophoblast cells, whereas anti-MRP1 monoclonal antibody MRPr1 stained the basal side. These results confirm that P-gp and MRP1 are located on the brush-border membranes and basal membranes, respectively, of human full-term placental trophoblast. MRP1 was also detected on the abluminal side of blood vessels in the villi. Accordingly, MRP1 may play a role distinct from that of P-gp, which is considered to restrict the influx of xenobiotics into the fetus.
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Affiliation(s)
- M Nagashige
- Department of Medico-Pharmaceutical Sciences, Graduate School of Pharmaceutical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, 812-8582 Fukuoka, Japan
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15
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Miranda CL, Stevens JF, Helmrich A, Henderson MC, Rodriguez RJ, Yang YH, Deinzer ML, Barnes DW, Buhler DR. Antiproliferative and cytotoxic effects of prenylated flavonoids from hops (Humulus lupulus) in human cancer cell lines. Food Chem Toxicol 1999; 37:271-85. [PMID: 10418944 DOI: 10.1016/s0278-6915(99)00019-8] [Citation(s) in RCA: 266] [Impact Index Per Article: 10.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/29/2022]
Abstract
Six flavonoids [xanthohumol (XN), 2',4',6',4-tetrahydroxy-3'-prenylchalcone (TP); 2',4',6',4-tetrahydroxy-3'-geranylchalcone (TG); dehydrocycloxanthohumol (DX); dehydrocycloxanthohumol hydrate (DH); and isoxanthohumol (IX)] from hops (Humulus lupulus) were tested for their antiproliferative activity in human breast cancer (MCF-7), colon cancer (HT-29) and ovarian cancer (A-2780) cells in vitro. XN, DX and IX caused a dose-dependent (0.1 to 100 microM) decrease in growth of all cancer cells. After a 2-day treatment, the concentrations at which the growth of MCF-7 cells was inhibited by 50% (IC50) were 13.3, 15.7 and 15.3 microM for XN, DX and IX, respectively. After a 4-day treatment, the IC50 for XN, DX and IX were 3.47, 6.87 and 4.69 microM, respectively. HT-29 cells were more resistant than MCF-7 cells to these flavonoids. In A-2780 cells, XN was highly antiproliferative with IC50 values of 0.52 and 5.2 microM after 2 and 4 days of exposure, respectively. At 100 microM, all the hop flavonoids were cytotoxic in the three cell lines. Growth inhibition of XN- and IX-treated MCF-7 cells was confirmed by cell counting. XN and IX inhibited DNA synthesis in MCF-7 cells. As antiproliferative agents, XN (chalcone) and IX (flavanone isomer of XN) may have potential chemopreventive activity against breast and ovarian cancer in humans.
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Affiliation(s)
- C L Miranda
- Department of Environmental and Molecular Toxicology, Oregon State University, Corvallis 97331, USA
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Hiss D, Gabriels G, Jacobs P, Folb P. Tunicamycin potentiates drug cytotoxicity and vincristine retention in multidrug resistant cell lines. Eur J Cancer 1996; 32A:2164-72. [PMID: 9014761 DOI: 10.1016/s0959-8049(96)00262-6] [Citation(s) in RCA: 8] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/03/2023]
Abstract
Tunicamycin (TM), an inhibitor of glycoprotein processing, was investigated for its potential to reverse the multiple drug resistance (MDR) phenotype. When TM was added in vitro to drug-resistant NIH-3T3-MDR and KB-8-5-11 cells, they developed an increased sensitivity to doxorubicin, epirubicin, vincristine and colchicine. Similarly, the sensitivity of NIH-3T3-MDR cells to cisplatin was also enhanced by TM. In the presence of TM, drug-sensitive NIH-3T3-parental cells exhibited greater susceptibility to the toxic effects of doxorubicin, epirubicin, vincristine (marginally significant), and colchicine, but not to cisplatin. Tunicamycin-treated drug-sensitive KB-3-1 cells showed an increased response to vincristine, but not to the other anticancer drugs. Pretreatment with TM inhibited glycoprotein synthesis in all the cell lines. Neither prior exposure to, nor co-incubation with TM, influenced the uptake of vincristine (VCR) in the various cell lines. However, NIH-3T3-MDR cells accumulated less VCR than their drug-sensitive controls and also exhibited reduced efflux of the drug when treated with TM. There were no significant differences in the levels of intracellular VCR uptake between drug-sensitive KB-3-1 and KB-8-5-11 cells. Tunicamycin increased intracellular VCR retention in KB-8-5-11 and NIH-3T3-MDR cells, but not in NIH-3T3-parental cells. However, drug-sensitive KB-3-1 cells expressed reduced VCR retention in response to TM exposure, indicating that correlations between VCR toxicity and its retention in the presence of TM should be made with caution. The results suggest that the enhancement of intracellular VCR retention in MDR cells lines caused by TM is likely to be the result of inhibition of VCR efflux. Inhibition of glycoprotein synthesis during TM exposure may account for the changes in VCR efflux and retention observed in the MDR cell lines. The enhancement of cisplatin cytotoxicity in NIH-3T3-MDR cells after exposure to TM is an interesting observation, since it is generally believed that agents which modify the MDR phenotype do not show a sensitising effect to cisplatin. These findings may have applications in the reversal of drug resistance.
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Affiliation(s)
- D Hiss
- Department of Pharmacology, University of Cape Town, Medical School, South Africa
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Hosoya KI, Kim KJ, Lee VH. Age-dependent expression of P-glycoprotein gp170 in Caco-2 cell monolayers. Pharm Res 1996; 13:885-90. [PMID: 8792427 DOI: 10.1023/a:1016005212640] [Citation(s) in RCA: 91] [Impact Index Per Article: 3.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/02/2023]
Abstract
PURPOSE To determine whether the expression and activity of the P-glycoprotein (P-GP) drug efflux pump vary with the culture age of Caco-2 cell monolayers. METHODS Caco-2 cell monolayers were grown for 3-27 days on tissue culture-treated Transwells. P-GP efflux function was determined by measuring transmonolayer fluxes of cyclosporin A (CsA) and verapamil, while P-GP expression level was evaluated by Western blot analysis using monoclonal antibody C219. RESULTS The apparent permeability coefficient (Papp) of CsA (0.5 microM) in the basolateral-to-apical (B-->A) direction increased with culture age and was higher than the apical-to-basolateral (A-->B) direction at all times. Net secretory Papp significantly increased from day 17 onward compared to that observed during day 3 through 13. Verapamil (100 microM) significantly inhibited CsA transport in the B-->A direction from day 17 to 27, while elevating CsA transport in the A-->B direction from day 6 to 27. Interestingly, the Papp of verapamil (0.5 microM) in the B-->A direction was significantly higher than in the A-->B direction from day 6 to 27, rendering increases in net secretory Papp of verapamil with culture age. Western analysis revealed that P-GP expression level was in the order of 4 weeks approximately 1 week > 3 weeks > 2 weeks at equal loading of cell proteins. CONCLUSIONS P-GP is continuously expressed throughout the culture period, but it may not be fully functional at an early age. Caco-2 cell monolayers of day 17 to 27 appear to be a good model to evaluate the functional role of P-GP in drug efflux.
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Affiliation(s)
- K I Hosoya
- University of Southern California, School of Pharmacy, Department of Pharmaceutical Sciences, Los Angeles 90033, USA
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