Ketoconazole Reverses Imatinib Resistance in Human Chronic Myelogenous Leukemia K562 Cells
Chronic myeloid leukemia (CML) is a hematologic disorder characterized by the oncogene BCR-ABL1, which encodes an oncoprotein with tyrosine kinase activity. Imatinib, a BCR-ABL1 tyrosine kinase inhibitor, performs exceptionally well with minimal toxicity in CML chemotherapy. According to clinical tr...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-07-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/23/14/7715 |
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| author | Omar Prado-Carrillo Abner Arenas-Ramírez Monserrat Llaguno-Munive Rafael Jurado Jazmin Pérez-Rojas Eduardo Cervera-Ceballos Patricia Garcia-Lopez |
| author_facet | Omar Prado-Carrillo Abner Arenas-Ramírez Monserrat Llaguno-Munive Rafael Jurado Jazmin Pérez-Rojas Eduardo Cervera-Ceballos Patricia Garcia-Lopez |
| author_sort | Omar Prado-Carrillo |
| collection | DOAJ |
| description | Chronic myeloid leukemia (CML) is a hematologic disorder characterized by the oncogene BCR-ABL1, which encodes an oncoprotein with tyrosine kinase activity. Imatinib, a BCR-ABL1 tyrosine kinase inhibitor, performs exceptionally well with minimal toxicity in CML chemotherapy. According to clinical trials, however, 20–30% of CML patients develop resistance to imatinib. Although the best studied resistance mechanisms are BCR-ABL1-dependent, P-glycoprotein (P-gp, a drug efflux transporter) may also contribute significantly. This study aimed to establish an imatinib-resistant human CML cell line, evaluate the role of P-gp in drug resistance, and assess the capacity of ketoconazole to reverse resistance by inhibiting P-gp. The following parameters were determined in both cell lines: cell viability (as the IC50) after exposure to imatinib and imatinib + ketoconazole, P-gp expression (by Western blot and immunofluorescence), the intracellular accumulation of a P-gp substrate (doxorubicin) by flow cytometry, and the percentage of apoptosis (by the Annexin method). In the highly resistant CML cell line obtained, P-gp was overexpressed, and the level of intracellular doxorubicin was low, representing high P-gp activity. Imatinib plus a non-toxic concentration of ketoconazole (10 μM) overcame drug resistance, inhibited P-gp overexpression and its efflux function, increased the intracellular accumulation of doxorubicin, and favored greater apoptosis of CML cells. P-gp contributes substantially to imatinib resistance in CML cells. Ketoconazole reversed CML cell resistance to imatinib by targeting P-gp-related pathways. The repurposing of ketoconazole for CML treatment will likely help patients resistant to imatinib. |
| first_indexed | 2024-03-09T13:42:27Z |
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| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-09T13:42:27Z |
| publishDate | 2022-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-8b9438f5b35342c98d1ed0688290d3df2023-11-30T21:05:01ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-07-012314771510.3390/ijms23147715Ketoconazole Reverses Imatinib Resistance in Human Chronic Myelogenous Leukemia K562 CellsOmar Prado-Carrillo0Abner Arenas-Ramírez1Monserrat Llaguno-Munive2Rafael Jurado3Jazmin Pérez-Rojas4Eduardo Cervera-Ceballos5Patricia Garcia-Lopez6Laboratorio de Fármaco-Oncología, Subdirección de Investigación Básica, Instituto Nacional de Cancerología, Ciudad de Mexico 14080, MexicoLaboratorio de Fármaco-Oncología, Subdirección de Investigación Básica, Instituto Nacional de Cancerología, Ciudad de Mexico 14080, MexicoLaboratorio de Fármaco-Oncología, Subdirección de Investigación Básica, Instituto Nacional de Cancerología, Ciudad de Mexico 14080, MexicoLaboratorio de Fármaco-Oncología, Subdirección de Investigación Básica, Instituto Nacional de Cancerología, Ciudad de Mexico 14080, MexicoLaboratorio de Fármaco-Oncología, Subdirección de Investigación Básica, Instituto Nacional de Cancerología, Ciudad de Mexico 14080, MexicoDirección de Docencia, Instituto Nacional de Cancerología, Ciudad de Mexico 14080, MexicoLaboratorio de Fármaco-Oncología, Subdirección de Investigación Básica, Instituto Nacional de Cancerología, Ciudad de Mexico 14080, MexicoChronic myeloid leukemia (CML) is a hematologic disorder characterized by the oncogene BCR-ABL1, which encodes an oncoprotein with tyrosine kinase activity. Imatinib, a BCR-ABL1 tyrosine kinase inhibitor, performs exceptionally well with minimal toxicity in CML chemotherapy. According to clinical trials, however, 20–30% of CML patients develop resistance to imatinib. Although the best studied resistance mechanisms are BCR-ABL1-dependent, P-glycoprotein (P-gp, a drug efflux transporter) may also contribute significantly. This study aimed to establish an imatinib-resistant human CML cell line, evaluate the role of P-gp in drug resistance, and assess the capacity of ketoconazole to reverse resistance by inhibiting P-gp. The following parameters were determined in both cell lines: cell viability (as the IC50) after exposure to imatinib and imatinib + ketoconazole, P-gp expression (by Western blot and immunofluorescence), the intracellular accumulation of a P-gp substrate (doxorubicin) by flow cytometry, and the percentage of apoptosis (by the Annexin method). In the highly resistant CML cell line obtained, P-gp was overexpressed, and the level of intracellular doxorubicin was low, representing high P-gp activity. Imatinib plus a non-toxic concentration of ketoconazole (10 μM) overcame drug resistance, inhibited P-gp overexpression and its efflux function, increased the intracellular accumulation of doxorubicin, and favored greater apoptosis of CML cells. P-gp contributes substantially to imatinib resistance in CML cells. Ketoconazole reversed CML cell resistance to imatinib by targeting P-gp-related pathways. The repurposing of ketoconazole for CML treatment will likely help patients resistant to imatinib.https://www.mdpi.com/1422-0067/23/14/7715chronic myeloid leukemiaimatinibtyrosine kinaseketoconazoleP-glycoproteindrug efflux transporter |
| spellingShingle | Omar Prado-Carrillo Abner Arenas-Ramírez Monserrat Llaguno-Munive Rafael Jurado Jazmin Pérez-Rojas Eduardo Cervera-Ceballos Patricia Garcia-Lopez Ketoconazole Reverses Imatinib Resistance in Human Chronic Myelogenous Leukemia K562 Cells International Journal of Molecular Sciences chronic myeloid leukemia imatinib tyrosine kinase ketoconazole P-glycoprotein drug efflux transporter |
| title | Ketoconazole Reverses Imatinib Resistance in Human Chronic Myelogenous Leukemia K562 Cells |
| title_full | Ketoconazole Reverses Imatinib Resistance in Human Chronic Myelogenous Leukemia K562 Cells |
| title_fullStr | Ketoconazole Reverses Imatinib Resistance in Human Chronic Myelogenous Leukemia K562 Cells |
| title_full_unstemmed | Ketoconazole Reverses Imatinib Resistance in Human Chronic Myelogenous Leukemia K562 Cells |
| title_short | Ketoconazole Reverses Imatinib Resistance in Human Chronic Myelogenous Leukemia K562 Cells |
| title_sort | ketoconazole reverses imatinib resistance in human chronic myelogenous leukemia k562 cells |
| topic | chronic myeloid leukemia imatinib tyrosine kinase ketoconazole P-glycoprotein drug efflux transporter |
| url | https://www.mdpi.com/1422-0067/23/14/7715 |
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