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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

Cross-resistance of sunitinib-resistant cells. (a) cross-resistance patterns, determined by MTT proliferation assays, of the sunitinib-resistant 786-O SUN and HT-29 SUN cell lines to sorafenib, pazopanib, erlotinib, lapatinib or everolimus compared to parental (PAR) cell lines. (b) intracellular accumulation of sorafenib, pazopanib or erlotinib in parental (PAR) and sunitinib-resistant (SUN) cells. Cells were incubated for 24 h with drug-containing medium at the ~ IC50 concentration of the parental cell line (except sunitinib itself). Drug concentrations: sunitinib: 5 μM (786-O) or 10 μM (HT-29); sorafenib: 2 μM (both cell lines); pazopanib: 4 μM (786-O) or 2 μM (HT-29); erlotinib: 5 μM (both cell lines). Results are shown as mean ± SEM (n = 2–3). *, p value < 0.05; **, p value < 0.01; ***, p value < 0.001
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Fig2: Cross-resistance of sunitinib-resistant cells. (a) cross-resistance patterns, determined by MTT proliferation assays, of the sunitinib-resistant 786-O SUN and HT-29 SUN cell lines to sorafenib, pazopanib, erlotinib, lapatinib or everolimus compared to parental (PAR) cell lines. (b) intracellular accumulation of sorafenib, pazopanib or erlotinib in parental (PAR) and sunitinib-resistant (SUN) cells. Cells were incubated for 24 h with drug-containing medium at the ~ IC50 concentration of the parental cell line (except sunitinib itself). Drug concentrations: sunitinib: 5 μM (786-O) or 10 μM (HT-29); sorafenib: 2 μM (both cell lines); pazopanib: 4 μM (786-O) or 2 μM (HT-29); erlotinib: 5 μM (both cell lines). Results are shown as mean ± SEM (n = 2–3). *, p value < 0.05; **, p value < 0.01; ***, p value < 0.001

Mentions: First, we confirmed our previously reported [7] sunitinib-resistance in the 786-O SUN and HT-29 SUN cell lines with a resistance factor (RF) of 3–4 fold (Fig. 2a and Table 1). Subsequently, we determined the sensitivity of 786-O SUN and HT-29 SUN cells to sorafenib, pazopanib, erlotinib, lapatinib and everolimus (Fig. 2a and Table 1). Erlotinib showed a pronounced cross-resistance in both 786-O SUN and HT-29 SUN cells (RF ≥ 4.5 and > 3.2, respectively) and did not reach an IC50 within the tested concentration range. Pazopanib showed cross-resistance in HT-29 SUN cells (RF = 22), but not in 786-O SUN cells (RF = 1.6). Two other TKIs, sorafenib and lapatinib, showed no cross-resistance in the sunitinib-resistant cells. The effect of everolimus on cell proliferation was modest over a wide concentration range in the parental cells, with no detectable cross-resistance in the sunitinib-resistant cell lines.Fig. 2


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)

Cross-resistance of sunitinib-resistant cells. (a) cross-resistance patterns, determined by MTT proliferation assays, of the sunitinib-resistant 786-O SUN and HT-29 SUN cell lines to sorafenib, pazopanib, erlotinib, lapatinib or everolimus compared to parental (PAR) cell lines. (b) intracellular accumulation of sorafenib, pazopanib or erlotinib in parental (PAR) and sunitinib-resistant (SUN) cells. Cells were incubated for 24 h with drug-containing medium at the ~ IC50 concentration of the parental cell line (except sunitinib itself). Drug concentrations: sunitinib: 5 μM (786-O) or 10 μM (HT-29); sorafenib: 2 μM (both cell lines); pazopanib: 4 μM (786-O) or 2 μM (HT-29); erlotinib: 5 μM (both cell lines). Results are shown as mean ± SEM (n = 2–3). *, p value < 0.05; **, p value < 0.01; ***, p value < 0.001
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Related In: Results  -  Collection

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Fig2: Cross-resistance of sunitinib-resistant cells. (a) cross-resistance patterns, determined by MTT proliferation assays, of the sunitinib-resistant 786-O SUN and HT-29 SUN cell lines to sorafenib, pazopanib, erlotinib, lapatinib or everolimus compared to parental (PAR) cell lines. (b) intracellular accumulation of sorafenib, pazopanib or erlotinib in parental (PAR) and sunitinib-resistant (SUN) cells. Cells were incubated for 24 h with drug-containing medium at the ~ IC50 concentration of the parental cell line (except sunitinib itself). Drug concentrations: sunitinib: 5 μM (786-O) or 10 μM (HT-29); sorafenib: 2 μM (both cell lines); pazopanib: 4 μM (786-O) or 2 μM (HT-29); erlotinib: 5 μM (both cell lines). Results are shown as mean ± SEM (n = 2–3). *, p value < 0.05; **, p value < 0.01; ***, p value < 0.001
Mentions: First, we confirmed our previously reported [7] sunitinib-resistance in the 786-O SUN and HT-29 SUN cell lines with a resistance factor (RF) of 3–4 fold (Fig. 2a and Table 1). Subsequently, we determined the sensitivity of 786-O SUN and HT-29 SUN cells to sorafenib, pazopanib, erlotinib, lapatinib and everolimus (Fig. 2a and Table 1). Erlotinib showed a pronounced cross-resistance in both 786-O SUN and HT-29 SUN cells (RF ≥ 4.5 and > 3.2, respectively) and did not reach an IC50 within the tested concentration range. Pazopanib showed cross-resistance in HT-29 SUN cells (RF = 22), but not in 786-O SUN cells (RF = 1.6). Two other TKIs, sorafenib and lapatinib, showed no cross-resistance in the sunitinib-resistant cells. The effect of everolimus on cell proliferation was modest over a wide concentration range in the parental cells, with no detectable cross-resistance in the sunitinib-resistant cell lines.Fig. 2

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