Fluorimetric Methods for Analysis of Permeability, Drug Transport Kinetics, and Inhibition of the ABCB1 Membrane Transporter

Armada, Ana; Martins, Célia; Spengler, Gabriella; Molnár, Joséph; Amaral, Leonard; Rodrigues, António Sebastião; Viveiros, Miguel

doi:10.1007/978-1-4939-3347-1_7

Cited by 11 publications

(10 citation statements)

References 41 publications

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“…The KCR cell line at time zero (week 0), overexpresses ABCB1 efflux pumps which are responsible for the drug resistance observed in this cell line. To evaluate the efflux activity of ABCB1, the DiOC 2 efflux assay was performed as previously described [20,29] and VP, a synthetic known inhibitor of ABCB1, was used to block efflux. DiOC 2 efflux was assessed through fluorescence microscopy and flow cytometry in KCR cells on week 0, 9 and 16, by exposing them to DiOC 2 with or without inhibitor (Figures 2 and 3).…”

Section: Loss Of Doxorubicin Resistance Is Accompanied By a Decrease mentioning

confidence: 99%

ABC Efflux Transporters and the Circuitry of miRNAs: Kinetics of Expression in Cancer Drug Resistance

Gomes

Honrado

Armada

et al. 2020

IJMS

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Cancer drug resistance (CDR) is a major problem in therapeutic failure. Over 90% of patients with metastatic cancer present CDR. Several mechanisms underlie CDR, including the increased expression of efflux ABC transporters and epigenetic phenomena. Nevertheless, a topic that is not usually addressed is the mechanism underlying the loss of CDR once the challenge to these cells is withdrawn. A KCR cell line (doxorubicin-resistant, expressing ABCB1) was used to induce loss of resistance by withdrawing doxorubicin in culture medium. ABCB1 activity was analysed by fluorescence microscopy and flow cytometry through substrate (DiOC2) retention assays. The expression of 1008 microRNAs was assessed before and after doxorubicin withdrawal. After 16 weeks of doxorubicin withdrawal, a decrease of ABCB1 activity and expression occurred. Moreover, we determined a signature of 23 microRNAs, 13 underexpressed and 10 overexpressed, as a tool to assess loss of resistance. Through pathway enrichment analysis, "Pathways in cancer", "Proteoglycans in cancer" and "ECM-receptor interaction" were identified as relevant in the loss of CDR. Taken together, the data reinforce the assumption that ABCB1 plays a major role in the kinetics of CDR, and their levels of expression are in the dependence of the circuitry of cell miRNAs.

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Section: Loss Of Doxorubicin Resistance Is Accompanied By a Decrease mentioning

confidence: 99%

ABC Efflux Transporters and the Circuitry of miRNAs: Kinetics of Expression in Cancer Drug Resistance

Gomes

Honrado

Armada

et al. 2020

IJMS

Self Cite

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“…Although treatment of this patient followed established protocols, it may be worth considering whether assessment of MDR1 and rhodamine 123 efflux should be included in patients not responding to steroid treatment. Verapamil or cyclosporine A can inhibit MDR1 and cyclosporin A was shown to improve responses to steroids in patients with steroid unresponsive chronic obstructive airways disease [ 34 ]. Corticosteroids are relatively non-specific in their effects and ideally more specific treatments would be preferred [ 17 , 35 ].…”

Section: Discussionmentioning

confidence: 99%

Anti-PD-1-induced high-grade hepatitis associated with corticosteroid-resistant T cells: a case report

McGuire

Shklovskaya

Edwards

et al. 2017

Cancer Immunol Immunother

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Effective treatment or prevention of immune side effects associated with checkpoint inhibitor therapy of cancer is an important goal in this new era of immunotherapy. Hepatitis due to immunotherapy with antibodies against PD-1 is uncommon and generally of low severity. We present an unusually severe case arising in a melanoma patient after more than 6 months uncomplicated treatment with anti-PD-1 in an adjuvant setting. The hepatitis rapidly developed resistance to high-dose steroids, requiring anti-thymocyte globulin (ATG) to achieve control. Mass cytometry allowed comprehensive phenotyping of circulating lymphocytes and revealed that CD4+ T cells were profoundly depleted by ATG, while CD8+ T cells, B cells, NK cells and monocytes were relatively spared. Multiple abnormalities in CD4+ T cell phenotype were stably present in the patient before disease onset. These included a population of CCR4−CCR6− effector/memory CD4+ T cells expressing intermediate levels of the Th1-related chemokine receptor CXCR3 and abnormally high multi-drug resistance type 1 transporter (MDR1) activity as assessed by a rhodamine 123 excretion assay. Expression of MDR1 has been implicated in steroid resistance and may have contributed to the severity and lack of a sustained steroid response in this patient. The number of CD4+ rhodamine 123-excreting cells was reduced > 3.5-fold after steroid and ATG treatment. This case illustrates the need to consider this form of steroid resistance in patients failing treatment with corticosteroids. It also highlights the need for both better identification of patients at risk and the development of treatments that involve more specific immune suppression.Electronic supplementary materialThe online version of this article (10.1007/s00262-017-2107-7) contains supplementary material, which is available to authorized users.

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“…Thus inhibition of this protein causes an increase in drug accumulation in multidrugresistant cells. 28 Another protein that was inhibited by Itraconazole is ALOX5 (Figure 3). The presence of inhibition in ALOX5 can cause the disruption of the inflammatory process.…”

Section: Hepatotoxicity Potentialmentioning

confidence: 99%

In silico analysis of antihypertensive and hepatotoxicity potential of the n-butanol fraction of the methanol extract of of cantaloupe (Cucumis melo var. cantalupensis)

Purwaningroom

Putri

Permatasari³

2020

hsji

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Latar belakang: Hipertensi merupakan faktor risiko utama penyakit kardiovaskular. Penduduk Indonesia cenderung mengkonsumsi herbal dalam terapi hipertensi dalam mempertahankan kadar tekanan darah seperti buah blewah (Cucumis melo var. cantalupensis). Namun mekanisme kerja buah blewah dalam menurunkan tekanan darah, dan potensi toksisitasnya jika dikonsumsi dalam jangka panjang masih belum jelas. Tujuan studi ini adalah untuk menganalisis mekanisme antihipertensi dari buah blewah dan potensi toksiknya melalui pendekatan in silico. Metode: Bubuk blewah kering dimaserasi menggunakan metanol absolut, difraksinasi menggunakan n-butanol. Uji fitokimia dilakukan dengan metode LC-MS, kemudian senyawa bioaktif ditelusuri hingga SMILESnya di PubChem. Analisis QSAR untuk analisis potensi antihipertensi dilakukan dengan PASS server. Kelas toksisitas dan potensi hepatotoksisitas dianalisis menggunakan ProTox-II, dilanjutkan dengan analisis networking menggunakan STITCH dan STRINGdb. Hasil: Setidaknya terdapat 434 jenis senyawa yang terdapat pada fraksi n-butanol dari ekstrak metanol buah blewah (FBEMB). Berdasarkan analisis STITCH dan STRINGdb, FBEMB dapat bekerja dalam menurunkan tekanan darah melalui mekanisme aksi seperti senyawa amlodipine, yang menstabilkan saluran kalsium tipe-L yang terisi tegangan dalam konformasi tidak aktifnya. Dengan demikian, mencegah kontraksi myocyte yang bergantung pada kalsium dan vasokonstriksi. FBEMB mungkin berpotensi hepatotoksik melalui mekanisme kerja senyawa seperti itrakonazol yang menghambat enzim sitokrom P450 yang mempengaruhi gangguan pada sintesis ergosterol, dan efavirenz yang memiliki efek neurotoksik. Penghambatan sitokrom P450 dapat menyebabkan toksisitas obat dan kerusakan hati. Kesimpulan: FBEMB dapat bekerja dalam menurunkan tekanan darah melalui mekanisme penstabilan saluran kalsium tipe-L yang terisi tegangan dalam konformasi tidak aktifnya. Kata kunci: in silico, antihipertensi, hepatotoksisitas, blewah Abstract Background: Hypertension is a major cardiovascular disease risk factor. Indonesian people tend to consume herbal medicine to maintain hypertension therapy, i.e cantaloupe (Cucumis melo var. cantalupensis). However, the mechanism of action of cantaloupe in lowering blood pressure and toxicity potential for long term consumption is unclear. The study aimed to analyze the antihypertensive mechanism of cantaloupe and its toxic potential through the in silico. Methods: The dried cantaloupe powder was macerated using absolute methanol, then fractionated using n-butanol. The phytochemical test was done by LC-MS method, then the bioactive compounds were traced to their SMILES in the PubChem. The QSAR analysis of the antihypertensive potential was done using the PASS server. The toxicity class and hepatotoxicity potential were analyzed using ProTox-II, followed by networking analysis using STITCH and STRINGdb. Results: At least 434 types of compounds were shown in the n-butanol fraction of the methanol extract of cantaloupe (BFMEC). Based on the networking analysis, BFMEC may work in lowering blood pressure through the action mechanism of the amlodipine compound-like, which stabilizes voltage-gated L-type calcium channels in an inactive conformation, thus, prevents calcium-dependent myocyte contraction and vasoconstriction. BFMEC presumably has hepatotoxic through the action mechanism of itraconazole compound-like inhibited cytochrome P450-dependent enzymes, affecting the impairment of ergosterol synthesis, and efavirenz which has neurotoxic effects. The inhibition of cytochrome P450 may cause drug toxicity and liver damage. Conclusion: BFMEC may work in lowering blood pressure through the action mechanism which stabilizes voltage-gated L-type calcium channels in an inactive conformation. Keywords: in silico, antihypertensive, hepatotoxicity, cantaloupe

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Fluorimetric Methods for Analysis of Permeability, Drug Transport Kinetics, and Inhibition of the ABCB1 Membrane Transporter

Cited by 11 publications

References 41 publications

ABC Efflux Transporters and the Circuitry of miRNAs: Kinetics of Expression in Cancer Drug Resistance

ABC Efflux Transporters and the Circuitry of miRNAs: Kinetics of Expression in Cancer Drug Resistance

Anti-PD-1-induced high-grade hepatitis associated with corticosteroid-resistant T cells: a case report

In silico analysis of antihypertensive and hepatotoxicity potential of the n-butanol fraction of the methanol extract of of cantaloupe (Cucumis melo var. cantalupensis)

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