Limits...
Alectinib (CH5424802) antagonizes ABCB1- and ABCG2-mediated multidrug resistance in vitro , in vivo and ex vivo

View Article: PubMed Central - PubMed

ABSTRACT

Alectinib, an inhibitor of anaplastic lymphoma kinase (ALK), was approved by the Food and Drug Administration (FDA) for the treatment of patients with ALK-positive non-small cell lung cancer (NSCLC). Here we investigated the reversal effect of alectinib on multidrug resistance (MDR) induced by ATP-binding cassette (ABC) transporters, which is the primary cause of chemotherapy failure. We provide the first evidence that alectinib increases the sensitivity of ABCB1- and ABCG2-overexpressing cells to chemotherapeutic agents in vitro and in vivo. Mechanistically, alectinib increased the intracellular accumulation of ABCB1/ABCG2 substrates such as doxorubicin (DOX) and Rhodamine 123 (Rho 123) by inhibiting the efflux function of the transporters in ABCB1- or ABCG2-overexpressing cells but not in their parental sensitive cells. Furthermore, alectinib stimulated ATPase activity and competed with substrates of ABCB1 or ABCG2 and competed with [125I] iodoarylazidoprazosin (IAAP) photolabeling bound to ABCB1 or ABCG2 but neither altered the expression and localization of ABCB1 or ABCG2 nor the phosphorylation levels of AKT and ERK. Alectinib also enhanced the cytotoxicity of DOX and the intracellular accumulation of Rho 123 in ABCB1-overexpressing primary leukemia cells. These findings suggest that alectinib combined with traditional chemotherapy may be beneficial to patients with ABCB1- or ABCG2-mediated MDR.

No MeSH data available.


The structure and cytotoxicity of alectinib. The structure of alectinib (a). The expression levels of ABCG2 and ABCB1 were detected in KB cells, ABCB1-overexpressing KBv200 cells, S1 cells and ABCG2-overexpressing S1-MI-80 cells (b). MTT assays were used to assess the cell survival in KB cells and ABCB1-overexpressing KBv200 cells (c), MCF-7 and ABCB1-overexpressing MCF-7/adr cells (d), S1 and ABCG2-overexpressing S1-MI-80 cells (e), H460 and ABCG2-overexpressing H460/MX20 cells (f), HEK293/Vector and stable ABCB1 transfected HEK293/ABCB1 cells (g), and HEK293/Vector and stable ABCG2 transfected HEK293/ABCG2-R2 cells (h). All cells were treated with a full range of alectinib concentrations for 72 h. The data points represent the mean±s.d. of at least three independent experiments performed in quadruple.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC5382559&req=5

fig1: The structure and cytotoxicity of alectinib. The structure of alectinib (a). The expression levels of ABCG2 and ABCB1 were detected in KB cells, ABCB1-overexpressing KBv200 cells, S1 cells and ABCG2-overexpressing S1-MI-80 cells (b). MTT assays were used to assess the cell survival in KB cells and ABCB1-overexpressing KBv200 cells (c), MCF-7 and ABCB1-overexpressing MCF-7/adr cells (d), S1 and ABCG2-overexpressing S1-MI-80 cells (e), H460 and ABCG2-overexpressing H460/MX20 cells (f), HEK293/Vector and stable ABCB1 transfected HEK293/ABCB1 cells (g), and HEK293/Vector and stable ABCG2 transfected HEK293/ABCG2-R2 cells (h). All cells were treated with a full range of alectinib concentrations for 72 h. The data points represent the mean±s.d. of at least three independent experiments performed in quadruple.

Mentions: Alectinib was purchased from KareBay Biochem (Monmouth Junction, NJ, USA). The formula of Alectinib is presented in Figure 1a. Antibodies against ABCB1/p-gp (sc-13131), ABCG2 (sc-377176), AKT (sc-8312), p-AKT (sc-7985-R), ERK (sc-514302) and p-ERK (sc-7383) were from Santa Cruz Biotechnology Inc. (Paso Robles, CA, USA). Flow cytometry antibodies against ABCB1 (#557002) and mouse IgG2b/κ (#559532) were purchased from BD Biosciences (San Jose, CA, USA), and ABCG2 (sc-18841) was from Santa Cruz Biotechnology Inc. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) antibody was from Kangchen Co. (Shanghai, China). SYBR Green qPCR Master Mix was from ExCell Bio (Shanghai, China). Dulbecco's modified Eagle's medium and RPMI 1640 medium were purchased from Thermo Fisher Scientific Inc. (Waltham, MA, USA). Rhodamine 123 (Rho 123), 1-(4,5-dimethylthiazol-2-yl)-3,5-diphenylformazan (MTT), G418, paclitaxel, doxorubicin (DOX), vincristine (VCR), verapamil (VRP), topotecan, mitoxantrone (MX), cisplatin, fumitremorgin C (FTC) and other chemicals were purchased from Sigma-Aldrich (St. Louis, MO, USA).


Alectinib (CH5424802) antagonizes ABCB1- and ABCG2-mediated multidrug resistance in vitro , in vivo and ex vivo
The structure and cytotoxicity of alectinib. The structure of alectinib (a). The expression levels of ABCG2 and ABCB1 were detected in KB cells, ABCB1-overexpressing KBv200 cells, S1 cells and ABCG2-overexpressing S1-MI-80 cells (b). MTT assays were used to assess the cell survival in KB cells and ABCB1-overexpressing KBv200 cells (c), MCF-7 and ABCB1-overexpressing MCF-7/adr cells (d), S1 and ABCG2-overexpressing S1-MI-80 cells (e), H460 and ABCG2-overexpressing H460/MX20 cells (f), HEK293/Vector and stable ABCB1 transfected HEK293/ABCB1 cells (g), and HEK293/Vector and stable ABCG2 transfected HEK293/ABCG2-R2 cells (h). All cells were treated with a full range of alectinib concentrations for 72 h. The data points represent the mean±s.d. of at least three independent experiments performed in quadruple.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5382559&req=5

fig1: The structure and cytotoxicity of alectinib. The structure of alectinib (a). The expression levels of ABCG2 and ABCB1 were detected in KB cells, ABCB1-overexpressing KBv200 cells, S1 cells and ABCG2-overexpressing S1-MI-80 cells (b). MTT assays were used to assess the cell survival in KB cells and ABCB1-overexpressing KBv200 cells (c), MCF-7 and ABCB1-overexpressing MCF-7/adr cells (d), S1 and ABCG2-overexpressing S1-MI-80 cells (e), H460 and ABCG2-overexpressing H460/MX20 cells (f), HEK293/Vector and stable ABCB1 transfected HEK293/ABCB1 cells (g), and HEK293/Vector and stable ABCG2 transfected HEK293/ABCG2-R2 cells (h). All cells were treated with a full range of alectinib concentrations for 72 h. The data points represent the mean±s.d. of at least three independent experiments performed in quadruple.
Mentions: Alectinib was purchased from KareBay Biochem (Monmouth Junction, NJ, USA). The formula of Alectinib is presented in Figure 1a. Antibodies against ABCB1/p-gp (sc-13131), ABCG2 (sc-377176), AKT (sc-8312), p-AKT (sc-7985-R), ERK (sc-514302) and p-ERK (sc-7383) were from Santa Cruz Biotechnology Inc. (Paso Robles, CA, USA). Flow cytometry antibodies against ABCB1 (#557002) and mouse IgG2b/κ (#559532) were purchased from BD Biosciences (San Jose, CA, USA), and ABCG2 (sc-18841) was from Santa Cruz Biotechnology Inc. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) antibody was from Kangchen Co. (Shanghai, China). SYBR Green qPCR Master Mix was from ExCell Bio (Shanghai, China). Dulbecco's modified Eagle's medium and RPMI 1640 medium were purchased from Thermo Fisher Scientific Inc. (Waltham, MA, USA). Rhodamine 123 (Rho 123), 1-(4,5-dimethylthiazol-2-yl)-3,5-diphenylformazan (MTT), G418, paclitaxel, doxorubicin (DOX), vincristine (VCR), verapamil (VRP), topotecan, mitoxantrone (MX), cisplatin, fumitremorgin C (FTC) and other chemicals were purchased from Sigma-Aldrich (St. Louis, MO, USA).

View Article: PubMed Central - PubMed

ABSTRACT

Alectinib, an inhibitor of anaplastic lymphoma kinase (ALK), was approved by the Food and Drug Administration (FDA) for the treatment of patients with ALK-positive non-small cell lung cancer (NSCLC). Here we investigated the reversal effect of alectinib on multidrug resistance (MDR) induced by ATP-binding cassette (ABC) transporters, which is the primary cause of chemotherapy failure. We provide the first evidence that alectinib increases the sensitivity of ABCB1- and ABCG2-overexpressing cells to chemotherapeutic agents in vitro and in vivo. Mechanistically, alectinib increased the intracellular accumulation of ABCB1/ABCG2 substrates such as doxorubicin (DOX) and Rhodamine 123 (Rho 123) by inhibiting the efflux function of the transporters in ABCB1- or ABCG2-overexpressing cells but not in their parental sensitive cells. Furthermore, alectinib stimulated ATPase activity and competed with substrates of ABCB1 or ABCG2 and competed with [125I] iodoarylazidoprazosin (IAAP) photolabeling bound to ABCB1 or ABCG2 but neither altered the expression and localization of ABCB1 or ABCG2 nor the phosphorylation levels of AKT and ERK. Alectinib also enhanced the cytotoxicity of DOX and the intracellular accumulation of Rho 123 in ABCB1-overexpressing primary leukemia cells. These findings suggest that alectinib combined with traditional chemotherapy may be beneficial to patients with ABCB1- or ABCG2-mediated MDR.

No MeSH data available.