ABCB1 AND SLC22A1 MRNA EXPRESSIONS ARE INCREASED IN A HOMOZYGOUS JAK2V617F CELL LINE TREATED WITH JAK INHIBITOR
(Abstract release date: 05/21/15)
EHA Library. Gomes G. 06/12/15; 102725; PB1901
Disclosure(s): Faculdade de Ciências Farmacêuticas, Universidade de São PauloDepartamento de Análises Clínicas e Toxicológicas
Guilherme Gomes
Contributions
Contributions
Abstract
Abstract: PB1901
Type: Publication Only
Background
Pharmacokinetics of many anticancer drugs is affected by drug transporters as ABCB1, ABCG2 and SLC22A1. These molecules are responsible for cell uptake (SLC22A1) or efflux (ABCB1 and ABCG2) of several xenobiotics and are widely distributed in many tissues. Several studies have shown their role in resistance and toxicity to chemotherapeutic agents. Dysregulation of JAK-STAT pathway is considered a hallmark of myeloproliferative neoplasms (MPN), and disruption of this pathway may result from mutations such as JAK2V617F. Therefore, therapies targeting the inhibition of JAK proteins have been developed. However, the impact of JAK-STAT pathway inhibition over ABCB1, ABCG2 and SLC22A1 gene expression in MPN cells has not been investigated yet.
Aims
To evaluate the effect of JAK-STAT pathway inhibition in the expression of drug transporters ABCB1, ABCG2 and SLC22A1 in JAK2V617F positive cell line HEL92.1.7.
Methods
Erythroleukemia homozygous JAK2V617F cell line HEL92.1.7 was treated with 1 µM of JAK Inhibitor I (Merck/Calbiochem, Darmstadt, Germany), an ATP-competitive inhibitor of JAK proteins, for 24h. Western blot was used for verifying JAK-STAT pathway inhibition by inactivation of STAT-5. Quantitative real time-PCR and flow cytometry analysis were used to assess ABCB1, ABCG2 and SLC22A1 mRNA and protein expression, respectively, in JAK inhibitor-treated cells and in vehicle-treated control cells.
Results
JAK-STAT pathway was inhibited by treatment with 1 µM of JAK Inhibitor I for 24h, confirmed by non-detection of phosphorylated STAT-5. ABCB1 and SLC22A1 mRNA expressions were increased after treatment (median of 1.58 and 1.64 fold change, respectively), however, no difference was observed between ABCB1, ABCG2 and SLC22A1 protein expression in cells treated with 1 µM of JAK Inhibitor I for 24h and controls (P>0.05 for all).
Summary
Our data suggest that JAK-STAT pathway inhibition could modulate mRNA expression of drug transporters ABCB1 and SLC22A1 in MPN cells. Further studies with longer periods of treatment may confirm this proposition and help to elucidate the effect of JAK-STAT pathway in ABCB1, ABCG2 and SLC22A1 protein expression.
Keyword(s): Cell line, Flow cytometry, Gene expression, Myeloproliferative disorder
Type: Publication Only
Background
Pharmacokinetics of many anticancer drugs is affected by drug transporters as ABCB1, ABCG2 and SLC22A1. These molecules are responsible for cell uptake (SLC22A1) or efflux (ABCB1 and ABCG2) of several xenobiotics and are widely distributed in many tissues. Several studies have shown their role in resistance and toxicity to chemotherapeutic agents. Dysregulation of JAK-STAT pathway is considered a hallmark of myeloproliferative neoplasms (MPN), and disruption of this pathway may result from mutations such as JAK2V617F. Therefore, therapies targeting the inhibition of JAK proteins have been developed. However, the impact of JAK-STAT pathway inhibition over ABCB1, ABCG2 and SLC22A1 gene expression in MPN cells has not been investigated yet.
Aims
To evaluate the effect of JAK-STAT pathway inhibition in the expression of drug transporters ABCB1, ABCG2 and SLC22A1 in JAK2V617F positive cell line HEL92.1.7.
Methods
Erythroleukemia homozygous JAK2V617F cell line HEL92.1.7 was treated with 1 µM of JAK Inhibitor I (Merck/Calbiochem, Darmstadt, Germany), an ATP-competitive inhibitor of JAK proteins, for 24h. Western blot was used for verifying JAK-STAT pathway inhibition by inactivation of STAT-5. Quantitative real time-PCR and flow cytometry analysis were used to assess ABCB1, ABCG2 and SLC22A1 mRNA and protein expression, respectively, in JAK inhibitor-treated cells and in vehicle-treated control cells.
Results
JAK-STAT pathway was inhibited by treatment with 1 µM of JAK Inhibitor I for 24h, confirmed by non-detection of phosphorylated STAT-5. ABCB1 and SLC22A1 mRNA expressions were increased after treatment (median of 1.58 and 1.64 fold change, respectively), however, no difference was observed between ABCB1, ABCG2 and SLC22A1 protein expression in cells treated with 1 µM of JAK Inhibitor I for 24h and controls (P>0.05 for all).
Summary
Our data suggest that JAK-STAT pathway inhibition could modulate mRNA expression of drug transporters ABCB1 and SLC22A1 in MPN cells. Further studies with longer periods of treatment may confirm this proposition and help to elucidate the effect of JAK-STAT pathway in ABCB1, ABCG2 and SLC22A1 protein expression.
Keyword(s): Cell line, Flow cytometry, Gene expression, Myeloproliferative disorder
Abstract: PB1901
Type: Publication Only
Background
Pharmacokinetics of many anticancer drugs is affected by drug transporters as ABCB1, ABCG2 and SLC22A1. These molecules are responsible for cell uptake (SLC22A1) or efflux (ABCB1 and ABCG2) of several xenobiotics and are widely distributed in many tissues. Several studies have shown their role in resistance and toxicity to chemotherapeutic agents. Dysregulation of JAK-STAT pathway is considered a hallmark of myeloproliferative neoplasms (MPN), and disruption of this pathway may result from mutations such as JAK2V617F. Therefore, therapies targeting the inhibition of JAK proteins have been developed. However, the impact of JAK-STAT pathway inhibition over ABCB1, ABCG2 and SLC22A1 gene expression in MPN cells has not been investigated yet.
Aims
To evaluate the effect of JAK-STAT pathway inhibition in the expression of drug transporters ABCB1, ABCG2 and SLC22A1 in JAK2V617F positive cell line HEL92.1.7.
Methods
Erythroleukemia homozygous JAK2V617F cell line HEL92.1.7 was treated with 1 µM of JAK Inhibitor I (Merck/Calbiochem, Darmstadt, Germany), an ATP-competitive inhibitor of JAK proteins, for 24h. Western blot was used for verifying JAK-STAT pathway inhibition by inactivation of STAT-5. Quantitative real time-PCR and flow cytometry analysis were used to assess ABCB1, ABCG2 and SLC22A1 mRNA and protein expression, respectively, in JAK inhibitor-treated cells and in vehicle-treated control cells.
Results
JAK-STAT pathway was inhibited by treatment with 1 µM of JAK Inhibitor I for 24h, confirmed by non-detection of phosphorylated STAT-5. ABCB1 and SLC22A1 mRNA expressions were increased after treatment (median of 1.58 and 1.64 fold change, respectively), however, no difference was observed between ABCB1, ABCG2 and SLC22A1 protein expression in cells treated with 1 µM of JAK Inhibitor I for 24h and controls (P>0.05 for all).
Summary
Our data suggest that JAK-STAT pathway inhibition could modulate mRNA expression of drug transporters ABCB1 and SLC22A1 in MPN cells. Further studies with longer periods of treatment may confirm this proposition and help to elucidate the effect of JAK-STAT pathway in ABCB1, ABCG2 and SLC22A1 protein expression.
Keyword(s): Cell line, Flow cytometry, Gene expression, Myeloproliferative disorder
Type: Publication Only
Background
Pharmacokinetics of many anticancer drugs is affected by drug transporters as ABCB1, ABCG2 and SLC22A1. These molecules are responsible for cell uptake (SLC22A1) or efflux (ABCB1 and ABCG2) of several xenobiotics and are widely distributed in many tissues. Several studies have shown their role in resistance and toxicity to chemotherapeutic agents. Dysregulation of JAK-STAT pathway is considered a hallmark of myeloproliferative neoplasms (MPN), and disruption of this pathway may result from mutations such as JAK2V617F. Therefore, therapies targeting the inhibition of JAK proteins have been developed. However, the impact of JAK-STAT pathway inhibition over ABCB1, ABCG2 and SLC22A1 gene expression in MPN cells has not been investigated yet.
Aims
To evaluate the effect of JAK-STAT pathway inhibition in the expression of drug transporters ABCB1, ABCG2 and SLC22A1 in JAK2V617F positive cell line HEL92.1.7.
Methods
Erythroleukemia homozygous JAK2V617F cell line HEL92.1.7 was treated with 1 µM of JAK Inhibitor I (Merck/Calbiochem, Darmstadt, Germany), an ATP-competitive inhibitor of JAK proteins, for 24h. Western blot was used for verifying JAK-STAT pathway inhibition by inactivation of STAT-5. Quantitative real time-PCR and flow cytometry analysis were used to assess ABCB1, ABCG2 and SLC22A1 mRNA and protein expression, respectively, in JAK inhibitor-treated cells and in vehicle-treated control cells.
Results
JAK-STAT pathway was inhibited by treatment with 1 µM of JAK Inhibitor I for 24h, confirmed by non-detection of phosphorylated STAT-5. ABCB1 and SLC22A1 mRNA expressions were increased after treatment (median of 1.58 and 1.64 fold change, respectively), however, no difference was observed between ABCB1, ABCG2 and SLC22A1 protein expression in cells treated with 1 µM of JAK Inhibitor I for 24h and controls (P>0.05 for all).
Summary
Our data suggest that JAK-STAT pathway inhibition could modulate mRNA expression of drug transporters ABCB1 and SLC22A1 in MPN cells. Further studies with longer periods of treatment may confirm this proposition and help to elucidate the effect of JAK-STAT pathway in ABCB1, ABCG2 and SLC22A1 protein expression.
Keyword(s): Cell line, Flow cytometry, Gene expression, Myeloproliferative disorder
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