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Cross-resistance to clinically used tyrosine kinase inhibitors sunitinib, sorafenib and pazopanib.

Gotink KJ, Rovithi M, de Haas RR, Honeywell RJ, Dekker H, Poel D, Azijli K, Peters GJ, Broxterman HJ, Verheul HM - Cell Oncol (Dordr) (2015)

Bottom Line: The previously generated sunitinib-resistant (SUN) renal cancer cells (786-O) and colorectal cancer cells (HT-29) were found to be cross-resistant to pazopanib, erlotinib and lapatinib, but not sorafenib.Intracellular drug accumulation was found to be increased in pazopanib- and erlotinib-, but not in sorafenib-exposed cells.Lysosomal capacity, reflected by LAMP1/2 expression, was found to be increased in resistant cells and, in addition, to be transient.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Oncology, VU University Medical Center, Rm 3A46, De Boelelaan 1117, 1081 HV, Amsterdam, The Netherlands.

ABSTRACT

Purpose: When during cancer treatment resistance to a tyrosine kinase inhibitor (TKI) occurs, switching to another TKI is often considered as a reasonable option. Previously, we reported that resistance to sunitinib may be caused by increased lysosomal sequestration, leading to increased intracellular lysosomal storage and, thereby, inactivity. Here, we studied the effect of several other TKIs on the development of (cross-) resistance.

Methods: TKI resistance was induced by continuous exposure of cancer cell lines to increasing TKI concentrations for 3-4 months. (Cross-) resistance was evaluated using MTT cell proliferation assays. Intracellular TKI concentrations were measured using LC-MS/MS. Western blotting was used to detect lysosome-associated membrane protein-1 and -2 (LAMP1/2) expression.

Results: The previously generated sunitinib-resistant (SUN) renal cancer cells (786-O) and colorectal cancer cells (HT-29) were found to be cross-resistant to pazopanib, erlotinib and lapatinib, but not sorafenib. Exposure of 786-O and HT-29 cells to sorafenib, pazopanib or erlotinib for 3-4 months induced drug resistance to pazopanib and erlotinib, but not sorafenib. Intracellular drug accumulation was found to be increased in pazopanib- and erlotinib-, but not in sorafenib-exposed cells. Lysosomal capacity, reflected by LAMP1/2 expression, was found to be increased in resistant cells and, in addition, to be transient. No cross-resistance to the mTOR inhibitor everolimus was detected.

Conclusions: Our data indicate that tumor cells can develop (cross-) resistance to TKIs, and that such resistance includes increased intracellular drug accumulation accompanied by increased lysosomal storage. Transient (cross-) resistance was found to occur for several of the TKIs tested, but not for everolimus, indicating that switching from a TKI to a mTOR inhibitor may be an attractive therapeutic option.

No MeSH data available.


Related in: MedlinePlus

Sensitivity of (parental) 786-O and HT-29 tumor cells to different inhibitors. Proliferation assays (MTT) of 786-O (left) and HT-29 (right) parental cell lines incubated with different concentrations of (a) the tyrosine kinase inhibitors (TKIs) sunitinib, sorafenib, pazopanib, erlotinib or lapatinib or (b) the mTOR inhibitor everolimus. Results are shown as mean ± SEM
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Fig1: Sensitivity of (parental) 786-O and HT-29 tumor cells to different inhibitors. Proliferation assays (MTT) of 786-O (left) and HT-29 (right) parental cell lines incubated with different concentrations of (a) the tyrosine kinase inhibitors (TKIs) sunitinib, sorafenib, pazopanib, erlotinib or lapatinib or (b) the mTOR inhibitor everolimus. Results are shown as mean ± SEM

Mentions: First, we determined the sensitivity of the parental cell lines 786-O PAR and HT-29 PAR to the tyrosine kinase inhibitors (TKIs) sunitinib, sorafenib and pazopanib, to the EGFR TKIs erlotinib and lapatinib and to the mTOR inhibitor everolimus in 96 h proliferation assays. The sensitivities to the respective TKIs were found to be in the same, low micro-molar range (Fig. 1a), with IC50 values between 0.8 and 6.5 μM (Table 1), and were comparable between the two cell lines. The mTOR inhibitor everolimus showed a different sensitivity curve compared to the TKIs and reached a plateau between ~1 nM and 10 μM, at which the proliferation hardly decreased (~IC60 (786-O) and ~ IC30 (HT29); Fig. 1b).Fig. 1


Cross-resistance to clinically used tyrosine kinase inhibitors sunitinib, sorafenib and pazopanib.

Gotink KJ, Rovithi M, de Haas RR, Honeywell RJ, Dekker H, Poel D, Azijli K, Peters GJ, Broxterman HJ, Verheul HM - Cell Oncol (Dordr) (2015)

Sensitivity of (parental) 786-O and HT-29 tumor cells to different inhibitors. Proliferation assays (MTT) of 786-O (left) and HT-29 (right) parental cell lines incubated with different concentrations of (a) the tyrosine kinase inhibitors (TKIs) sunitinib, sorafenib, pazopanib, erlotinib or lapatinib or (b) the mTOR inhibitor everolimus. Results are shown as mean ± SEM
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4555235&req=5

Fig1: Sensitivity of (parental) 786-O and HT-29 tumor cells to different inhibitors. Proliferation assays (MTT) of 786-O (left) and HT-29 (right) parental cell lines incubated with different concentrations of (a) the tyrosine kinase inhibitors (TKIs) sunitinib, sorafenib, pazopanib, erlotinib or lapatinib or (b) the mTOR inhibitor everolimus. Results are shown as mean ± SEM
Mentions: First, we determined the sensitivity of the parental cell lines 786-O PAR and HT-29 PAR to the tyrosine kinase inhibitors (TKIs) sunitinib, sorafenib and pazopanib, to the EGFR TKIs erlotinib and lapatinib and to the mTOR inhibitor everolimus in 96 h proliferation assays. The sensitivities to the respective TKIs were found to be in the same, low micro-molar range (Fig. 1a), with IC50 values between 0.8 and 6.5 μM (Table 1), and were comparable between the two cell lines. The mTOR inhibitor everolimus showed a different sensitivity curve compared to the TKIs and reached a plateau between ~1 nM and 10 μM, at which the proliferation hardly decreased (~IC60 (786-O) and ~ IC30 (HT29); Fig. 1b).Fig. 1

Bottom Line: The previously generated sunitinib-resistant (SUN) renal cancer cells (786-O) and colorectal cancer cells (HT-29) were found to be cross-resistant to pazopanib, erlotinib and lapatinib, but not sorafenib.Intracellular drug accumulation was found to be increased in pazopanib- and erlotinib-, but not in sorafenib-exposed cells.Lysosomal capacity, reflected by LAMP1/2 expression, was found to be increased in resistant cells and, in addition, to be transient.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Oncology, VU University Medical Center, Rm 3A46, De Boelelaan 1117, 1081 HV, Amsterdam, The Netherlands.

ABSTRACT

Purpose: When during cancer treatment resistance to a tyrosine kinase inhibitor (TKI) occurs, switching to another TKI is often considered as a reasonable option. Previously, we reported that resistance to sunitinib may be caused by increased lysosomal sequestration, leading to increased intracellular lysosomal storage and, thereby, inactivity. Here, we studied the effect of several other TKIs on the development of (cross-) resistance.

Methods: TKI resistance was induced by continuous exposure of cancer cell lines to increasing TKI concentrations for 3-4 months. (Cross-) resistance was evaluated using MTT cell proliferation assays. Intracellular TKI concentrations were measured using LC-MS/MS. Western blotting was used to detect lysosome-associated membrane protein-1 and -2 (LAMP1/2) expression.

Results: The previously generated sunitinib-resistant (SUN) renal cancer cells (786-O) and colorectal cancer cells (HT-29) were found to be cross-resistant to pazopanib, erlotinib and lapatinib, but not sorafenib. Exposure of 786-O and HT-29 cells to sorafenib, pazopanib or erlotinib for 3-4 months induced drug resistance to pazopanib and erlotinib, but not sorafenib. Intracellular drug accumulation was found to be increased in pazopanib- and erlotinib-, but not in sorafenib-exposed cells. Lysosomal capacity, reflected by LAMP1/2 expression, was found to be increased in resistant cells and, in addition, to be transient. No cross-resistance to the mTOR inhibitor everolimus was detected.

Conclusions: Our data indicate that tumor cells can develop (cross-) resistance to TKIs, and that such resistance includes increased intracellular drug accumulation accompanied by increased lysosomal storage. Transient (cross-) resistance was found to occur for several of the TKIs tested, but not for everolimus, indicating that switching from a TKI to a mTOR inhibitor may be an attractive therapeutic option.

No MeSH data available.


Related in: MedlinePlus