Chemosensitization and drug accumulation assays as complementary methods for the screening of multidrug resistance reversal agents

Quesada, Ana-Rodriguez; Barbacid, M.M.; Mira, Emilia; Aracil, Miguel; Márquez, Gabriel

doi:10.1016/0304-3835(95)04044-7

Cited by 16 publications

(9 citation statements)

References 14 publications

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“…The small increase in non-MDR cell accumulation of H33342 with reserpine is likely due to nonspecific interactions between reserpine and cellular components unrelated to Pgp. The finding that R123 offered a more sensitive measure of Pgp function than R6G agrees with previous microplate screening results, 21 but contrasts with one flow cytometry assay that found R6G to give a greater difference than R123 between MDR and non-MDR cells. 17 The greater sensitivity of R123 over H33342 is in agreement with several flow cytometry studies.…”

Section: Protein Assay Considerationssupporting

confidence: 86%

“…R123 as well as the related rhodamine 6G (R6G) have been employed in microplate-based screening assays of cellular Pgp function. [20][21][22] These cellular assays have sought to ascertain general Pgp function or to evaluate overall chemosensitizing agent activity, however, and have not been designed with the intent of distinguishing interactions with the distinct Pgp binding sites.…”

Section: Introductionmentioning

confidence: 99%

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Microplate Screening of the Differential Effects of Test Agents on Hoechst 33342, Rhodamine 123, and Rhodamine 6G Accumulation in Breast Cancer Cells that Overexpress P–Glycoprotein

Sarver¹,

Kliś²,

Byers³

et al. 2002

SLAS Discovery

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A microplate screening method has been developed to evaluate the effects of test agents on the accumulation of the fluorescent P-glycoprotein (Pgp) substrates Hoechst 33342, rhodamine 123, and rhodamine 6G in multidrug-resistant (MDR) breast cancer cells that overexpress Pgp. All three substrates exhibit substantially higher accumulation in MCF7 non-MDR cells versus NCI/ADR-RES MDR cells, while incubation with 50 microM reserpine significantly reduces or eliminates these differences. Rhodamine 123 shows the lowest substrate accumulation efficiency in non-MDR cells relative to the substrate incubation level. The effects of several chemosensitizing agents and a series of paclitaxel analogs on the accumulation of each fluorescent substrate suggest that there are distinct differences in the substrate interaction profiles exhibited by these different agents. The described methods may be useful in Pgp-related research in the areas of cancer MDR, oral drug absorption, the blood-brain barrier, renal/hepatic transport processes, and drug-drug interactions.

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Section: Protein Assay Considerationssupporting

confidence: 86%

Section: Introductionmentioning

confidence: 99%

Microplate Screening of the Differential Effects of Test Agents on Hoechst 33342, Rhodamine 123, and Rhodamine 6G Accumulation in Breast Cancer Cells that Overexpress P–Glycoprotein

Sarver¹,

Kliś²,

Byers³

et al. 2002

SLAS Discovery

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“…[70][71][72][73] Collateral sensitivity has been demonstrated for drug concentrations in the Figure 4 Continued micromolar range. 72,74 Conversely, the selective cytotoxicity of the modulators in this study appeared at considerably lower concentrations, which facilitates possible therapeutic exploitation, and resulted specifically in cytokinesis failure and apoptosis induction. The maximally tolerable plasma concentration of PSC 833 in cancer patients is 2.5-3 m, 19 which demonstrates that the concentrations applied in our study are clinically relevant.…”

Section: Discussionmentioning

confidence: 80%

Growth inhibition, cytokinesis failure and apoptosis of multidrug-resistant leukemia cells after treatment with P-glycoprotein inhibitory agents

et al. 1999

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The multidrug transporter P-glycoprotein (Pgp), which is frequently overexpressed in multidrug resistant leukemia, has many proposed physiological functions including involvement in transmembraneous transport of certain growth-regulating cytokines. Therefore, we studied cell growth of three pairs of drug resistant and sensitive leukemia cell lines (KG1a, K562 and HL60) exposed to three different inhibitors of Pgp. The resistant KG1a and K562 sublines, which expressed high levels of Pgp, responded to low doses of the cyclosporin SDZ PSC 833, the cyclopeptolide SDZ 280-446, and the cyclopropyldibenzosuberane LY335979 with a dose-dependent growth inhibition. In the resistant variants of KG1a and K562 cells the mean half-maximal growth inhibitory doses (GI 50 ) of SDZ PSC 833 were 312 (SE 41) and 414 (SE 50) nM, those of SDZ 280-446 were 685 (SE 51) and 578 (SE 54) nM, and those of LY335979 were 66 (SE 1) and 48 (SE 8) nM, respectively. Exposure to 1 M SDZ PSC 833 resulted in tetraploidization, cytokinesis failure and apoptosis of the KG1a and K562 MDR variants. Conversely, parental cells with no or low levels of Pgp and the non-Pgp resistant variant of HL60 cells were not receptive to these cytotoxic effects. We conclude that inhibition of Pgp may exercise selective cytotoxicity in Pgp-rich leukemia cells indicating a possible therapeutic target in multiresistant leukemia.

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“…After establishing the maximum non-cytotoxic concentration for each compound, the minimum MDR reversal concentration was found by means of a cytotoxicity assay that included non-toxic concentrations of the compound and a fixed concentration of doxorubicin [16]. Cells (2,000 cells per well in a final volume of 100 µl) were incubated in each well with serial dilutions of the compound to be assayed and a fixed concentration (0.4 µ M) of doxorubicin, which is not toxic for Schabel cells.…”

Section: Mdr Reversal Assaysmentioning

confidence: 99%

MDR Reversal by Deprenylated Tetracyclic and Hexacyclic Analogues of N-Acetylardeemin: Confirmation of the Ardeemin Pharmacophore

Avendaño¹,

Caballero²,

Méndez-Vidal³

et al. 2006

LDDD

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H3C H H R = CH 3 N-Acetylardeemin (1a) R = H (1b) AV-200 (2) Fumitrem orgin C (3) Fig. (1). Structure of N-acetylardeemin and related compounds. Abstract:The MDR chemosensitising activity of several analogues of the ardeemins, including five derivatives of the ABCD fragment, nine derivatives of the complete hexacyclic framework and one seco analogue lacking the B ring was studied. The results obtained confirm that the pharmacophoric moiety of the ardeemins as MDR reversers is located at their DEF fragment, and suggest that the epimer of the ardeemins at the alanine stereocenter should be more active than the natural stereoisomer.

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Chemosensitization and drug accumulation assays as complementary methods for the screening of multidrug resistance reversal agents

Cited by 16 publications

References 14 publications

Microplate Screening of the Differential Effects of Test Agents on Hoechst 33342, Rhodamine 123, and Rhodamine 6G Accumulation in Breast Cancer Cells that Overexpress P–Glycoprotein

Microplate Screening of the Differential Effects of Test Agents on Hoechst 33342, Rhodamine 123, and Rhodamine 6G Accumulation in Breast Cancer Cells that Overexpress P–Glycoprotein

Growth inhibition, cytokinesis failure and apoptosis of multidrug-resistant leukemia cells after treatment with P-glycoprotein inhibitory agents

MDR Reversal by Deprenylated Tetracyclic and Hexacyclic Analogues of N-Acetylardeemin: Confirmation of the Ardeemin Pharmacophore

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