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Tyrosine kinase chromosomal translocations mediate distinct and overlapping gene regulation events.

Kim H, Gillis LC, Jarvis JD, Yang S, Huang K, Der S, Barber DL - BMC Cancer (2011)

Bottom Line: Our results suggest that BCR-ABL, TEL-PDGFRB and TEL-JAK2 regulate distinct and overlapping gene transcription profiles.Many of the genes identified are known to be involved in processes associated with leukemogenesis, including cell migration, proliferation and differentiation.This study offers the basis for further work that could lead to an understanding of the specificity of diseases caused by these three chromosomal translocations.

View Article: PubMed Central - HTML - PubMed

Affiliation: Campbell Family Cancer Research Institute, Ontario Cancer Institute, University Health Network, 610 University Avenue, Toronto, Ontario M5G 2M9, Canada.

ABSTRACT

Background: Leukemia is a heterogeneous disease commonly associated with recurrent chromosomal translocations that involve tyrosine kinases including BCR-ABL, TEL-PDGFRB and TEL-JAK2. Most studies on the activated tyrosine kinases have focused on proximal signaling events, but little is known about gene transcription regulated by these fusions.

Methods: Oligonucleotide microarray was performed to compare mRNA changes attributable to BCR-ABL, TEL-PDGFRB and TEL-JAK2 after 1 week of activation of each fusion in Ba/F3 cell lines. Imatinib was used to control the activation of BCR-ABL and TEL-PDGFRB, and TEL-JAK2-mediated gene expression was examined 1 week after Ba/F3-TEL-JAK2 cells were switched to factor-independent conditions.

Results: Microarray analysis revealed between 800 to 2000 genes induced or suppressed by two-fold or greater by each tyrosine kinase, with a subset of these genes commonly induced or suppressed among the three fusions. Validation by Quantitative PCR confirmed that eight genes (Dok2, Mrvi1, Isg20, Id1, gp49b, Cxcl10, Scinderin, and collagen Vα1(Col5a1)) displayed an overlapping regulation among the three tested fusion proteins. Stat1 and Gbp1 were induced uniquely by TEL-PDGFRB.

Conclusions: Our results suggest that BCR-ABL, TEL-PDGFRB and TEL-JAK2 regulate distinct and overlapping gene transcription profiles. Many of the genes identified are known to be involved in processes associated with leukemogenesis, including cell migration, proliferation and differentiation. This study offers the basis for further work that could lead to an understanding of the specificity of diseases caused by these three chromosomal translocations.

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Related in: MedlinePlus

A subset of genes is commonly induced by BCR-ABL, TEL-PDGFRB and TEL-JAK2. Ba/F3 BCR-ABL cells and Ba/F3 TEL-PDGFRB cells were washed and incubated in the media depleted of IL-3 and supplemented with Imatinib for 5 h. Cells were washed and incubated in the absence of IL-3 and Imatinib to activate the fusion kinases for 0 h and 1 week. Ba/F3 TEL-JAK2 cells were washed and incubated in IL-3-depleted media for 1 week, and Ba/F3 cells were washed and incubated in IL-3-depleted media for 5 h to be used as a reference for Ba/F3 TEL-JAK2 cells. Total RNA was collected from each cell line and microarray analysis was performed using a mouse oligonucleotide array. For cells expressing BCR-ABL or TEL-PDGFRB, changes in gene expression were calculated using the expression values at 1 week and 0 h. For Ba/F3 TEL-JAK2 cells, expression values obtained from Ba/F3 TEL-JAK2 cells at the 1-week time point were compared to the expression values obtained from Ba/F3 cells after 5 h of IL3-depletion. Fold-inductions of gene expression are indicated in the table (Bottom) and are displayed in a bar-graph (Top). Asterisks denotes genes that were validated by Q-PCR. Phlda1, Pleckstrin homology-like domain, family A, member 1; Pdxl, podocalyxin; Dusp6, dual specificity phosphatase 6; Ifi47, interferon gamma inducible protein, 47 kDa; Il4ra, interleukin 4 receptor, alpha; Tcrg-V4, T-cell receptor gamma, variable 4; Pim1, proviral integration site 1; Cish, cytokine inducible SH2-containing protein; Isg20, interferon stimulated protein 20; gp49b, C3H gp49b; Spred1, Sprouty protein with EVH-1 domain 1, related sequence; Id1, transcription factor Id1; Atf4, activating transcription factor 4; Siah2, Seven in absentia 2; Serpinb1a, serine(or cysteine) proteinase inhibitor, clade B (ovalbumin) 1a.
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Figure 3: A subset of genes is commonly induced by BCR-ABL, TEL-PDGFRB and TEL-JAK2. Ba/F3 BCR-ABL cells and Ba/F3 TEL-PDGFRB cells were washed and incubated in the media depleted of IL-3 and supplemented with Imatinib for 5 h. Cells were washed and incubated in the absence of IL-3 and Imatinib to activate the fusion kinases for 0 h and 1 week. Ba/F3 TEL-JAK2 cells were washed and incubated in IL-3-depleted media for 1 week, and Ba/F3 cells were washed and incubated in IL-3-depleted media for 5 h to be used as a reference for Ba/F3 TEL-JAK2 cells. Total RNA was collected from each cell line and microarray analysis was performed using a mouse oligonucleotide array. For cells expressing BCR-ABL or TEL-PDGFRB, changes in gene expression were calculated using the expression values at 1 week and 0 h. For Ba/F3 TEL-JAK2 cells, expression values obtained from Ba/F3 TEL-JAK2 cells at the 1-week time point were compared to the expression values obtained from Ba/F3 cells after 5 h of IL3-depletion. Fold-inductions of gene expression are indicated in the table (Bottom) and are displayed in a bar-graph (Top). Asterisks denotes genes that were validated by Q-PCR. Phlda1, Pleckstrin homology-like domain, family A, member 1; Pdxl, podocalyxin; Dusp6, dual specificity phosphatase 6; Ifi47, interferon gamma inducible protein, 47 kDa; Il4ra, interleukin 4 receptor, alpha; Tcrg-V4, T-cell receptor gamma, variable 4; Pim1, proviral integration site 1; Cish, cytokine inducible SH2-containing protein; Isg20, interferon stimulated protein 20; gp49b, C3H gp49b; Spred1, Sprouty protein with EVH-1 domain 1, related sequence; Id1, transcription factor Id1; Atf4, activating transcription factor 4; Siah2, Seven in absentia 2; Serpinb1a, serine(or cysteine) proteinase inhibitor, clade B (ovalbumin) 1a.

Mentions: Finally, we identified a subset of genes that were regulated by all three fusion kinases. Id1, gp49b, Col5a1, Scinderin and Isg20 were selected for validation by Q-PCR (Figures 3 &4). Among the five transcripts, only Scinderin was commonly suppressed by all three fusions in the Q-PCR analysis, while the other four transcripts displayed overlapping regulation between two of the three fusion kinases (Table 5).


Tyrosine kinase chromosomal translocations mediate distinct and overlapping gene regulation events.

Kim H, Gillis LC, Jarvis JD, Yang S, Huang K, Der S, Barber DL - BMC Cancer (2011)

A subset of genes is commonly induced by BCR-ABL, TEL-PDGFRB and TEL-JAK2. Ba/F3 BCR-ABL cells and Ba/F3 TEL-PDGFRB cells were washed and incubated in the media depleted of IL-3 and supplemented with Imatinib for 5 h. Cells were washed and incubated in the absence of IL-3 and Imatinib to activate the fusion kinases for 0 h and 1 week. Ba/F3 TEL-JAK2 cells were washed and incubated in IL-3-depleted media for 1 week, and Ba/F3 cells were washed and incubated in IL-3-depleted media for 5 h to be used as a reference for Ba/F3 TEL-JAK2 cells. Total RNA was collected from each cell line and microarray analysis was performed using a mouse oligonucleotide array. For cells expressing BCR-ABL or TEL-PDGFRB, changes in gene expression were calculated using the expression values at 1 week and 0 h. For Ba/F3 TEL-JAK2 cells, expression values obtained from Ba/F3 TEL-JAK2 cells at the 1-week time point were compared to the expression values obtained from Ba/F3 cells after 5 h of IL3-depletion. Fold-inductions of gene expression are indicated in the table (Bottom) and are displayed in a bar-graph (Top). Asterisks denotes genes that were validated by Q-PCR. Phlda1, Pleckstrin homology-like domain, family A, member 1; Pdxl, podocalyxin; Dusp6, dual specificity phosphatase 6; Ifi47, interferon gamma inducible protein, 47 kDa; Il4ra, interleukin 4 receptor, alpha; Tcrg-V4, T-cell receptor gamma, variable 4; Pim1, proviral integration site 1; Cish, cytokine inducible SH2-containing protein; Isg20, interferon stimulated protein 20; gp49b, C3H gp49b; Spred1, Sprouty protein with EVH-1 domain 1, related sequence; Id1, transcription factor Id1; Atf4, activating transcription factor 4; Siah2, Seven in absentia 2; Serpinb1a, serine(or cysteine) proteinase inhibitor, clade B (ovalbumin) 1a.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC3295743&req=5

Figure 3: A subset of genes is commonly induced by BCR-ABL, TEL-PDGFRB and TEL-JAK2. Ba/F3 BCR-ABL cells and Ba/F3 TEL-PDGFRB cells were washed and incubated in the media depleted of IL-3 and supplemented with Imatinib for 5 h. Cells were washed and incubated in the absence of IL-3 and Imatinib to activate the fusion kinases for 0 h and 1 week. Ba/F3 TEL-JAK2 cells were washed and incubated in IL-3-depleted media for 1 week, and Ba/F3 cells were washed and incubated in IL-3-depleted media for 5 h to be used as a reference for Ba/F3 TEL-JAK2 cells. Total RNA was collected from each cell line and microarray analysis was performed using a mouse oligonucleotide array. For cells expressing BCR-ABL or TEL-PDGFRB, changes in gene expression were calculated using the expression values at 1 week and 0 h. For Ba/F3 TEL-JAK2 cells, expression values obtained from Ba/F3 TEL-JAK2 cells at the 1-week time point were compared to the expression values obtained from Ba/F3 cells after 5 h of IL3-depletion. Fold-inductions of gene expression are indicated in the table (Bottom) and are displayed in a bar-graph (Top). Asterisks denotes genes that were validated by Q-PCR. Phlda1, Pleckstrin homology-like domain, family A, member 1; Pdxl, podocalyxin; Dusp6, dual specificity phosphatase 6; Ifi47, interferon gamma inducible protein, 47 kDa; Il4ra, interleukin 4 receptor, alpha; Tcrg-V4, T-cell receptor gamma, variable 4; Pim1, proviral integration site 1; Cish, cytokine inducible SH2-containing protein; Isg20, interferon stimulated protein 20; gp49b, C3H gp49b; Spred1, Sprouty protein with EVH-1 domain 1, related sequence; Id1, transcription factor Id1; Atf4, activating transcription factor 4; Siah2, Seven in absentia 2; Serpinb1a, serine(or cysteine) proteinase inhibitor, clade B (ovalbumin) 1a.
Mentions: Finally, we identified a subset of genes that were regulated by all three fusion kinases. Id1, gp49b, Col5a1, Scinderin and Isg20 were selected for validation by Q-PCR (Figures 3 &4). Among the five transcripts, only Scinderin was commonly suppressed by all three fusions in the Q-PCR analysis, while the other four transcripts displayed overlapping regulation between two of the three fusion kinases (Table 5).

Bottom Line: Our results suggest that BCR-ABL, TEL-PDGFRB and TEL-JAK2 regulate distinct and overlapping gene transcription profiles.Many of the genes identified are known to be involved in processes associated with leukemogenesis, including cell migration, proliferation and differentiation.This study offers the basis for further work that could lead to an understanding of the specificity of diseases caused by these three chromosomal translocations.

View Article: PubMed Central - HTML - PubMed

Affiliation: Campbell Family Cancer Research Institute, Ontario Cancer Institute, University Health Network, 610 University Avenue, Toronto, Ontario M5G 2M9, Canada.

ABSTRACT

Background: Leukemia is a heterogeneous disease commonly associated with recurrent chromosomal translocations that involve tyrosine kinases including BCR-ABL, TEL-PDGFRB and TEL-JAK2. Most studies on the activated tyrosine kinases have focused on proximal signaling events, but little is known about gene transcription regulated by these fusions.

Methods: Oligonucleotide microarray was performed to compare mRNA changes attributable to BCR-ABL, TEL-PDGFRB and TEL-JAK2 after 1 week of activation of each fusion in Ba/F3 cell lines. Imatinib was used to control the activation of BCR-ABL and TEL-PDGFRB, and TEL-JAK2-mediated gene expression was examined 1 week after Ba/F3-TEL-JAK2 cells were switched to factor-independent conditions.

Results: Microarray analysis revealed between 800 to 2000 genes induced or suppressed by two-fold or greater by each tyrosine kinase, with a subset of these genes commonly induced or suppressed among the three fusions. Validation by Quantitative PCR confirmed that eight genes (Dok2, Mrvi1, Isg20, Id1, gp49b, Cxcl10, Scinderin, and collagen Vα1(Col5a1)) displayed an overlapping regulation among the three tested fusion proteins. Stat1 and Gbp1 were induced uniquely by TEL-PDGFRB.

Conclusions: Our results suggest that BCR-ABL, TEL-PDGFRB and TEL-JAK2 regulate distinct and overlapping gene transcription profiles. Many of the genes identified are known to be involved in processes associated with leukemogenesis, including cell migration, proliferation and differentiation. This study offers the basis for further work that could lead to an understanding of the specificity of diseases caused by these three chromosomal translocations.

Show MeSH
Related in: MedlinePlus