Expression of the cholecystokinin gene in pediatric tumors.

Friedman, Jeffrey M.; Vitale, María; Maimon, Jonathan; Israel, Mark A.; Horowitz, Marc E.; Schneider, B

doi:10.1073/pnas.89.13.5819

Cited by 16 publications

(9 citation statements)

References 34 publications

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“…Our data show that CCK is overexpressed in Ewing tumors, with levels similar to those detected in small intestine and fetal brain (the two normal tissues tested with the highest levels of CCK) and that CCK can be considered as a Ewing specific marker, at least when compared with other pediatric malignancies such as neuroblastoma and rhabdomyosarcoma. Our studies extend those of Friedman et al (29) and Schneider et al (41) …”

Section: Discussionsupporting

confidence: 79%

Cholecystokinin Down-Regulation by RNA Interference Impairs Ewing Tumor Growth

Carrillo

García-Aragoncillo

Azorín

et al. 2007

Clinical Cancer Research

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Purpose:Tumors of the Ewing family are characterized by chromosomal translocations that yield chimeric transcription factors, such as EWS/FLI1, which regulate the expression of specific genes that contribute to the malignant phenotype. In the present study, we show that cholecystokinin (CCK) is a new target of the EWS/FLI1oncoprotein and assess its functional role in Ewing tumor pathogenesis. Experimental Design: Relevant EWS/FLI1 targets were identified using a combination of cell systems with inducible EWS/FLI1expression, Ewing tumors and cell lines, microarrays, and RNA interference with doxycycline-inducible small hairpin RNA (shRNA) vectors. A doxycyclineinducible CCK-shRNA vector was stably transfected in A673 and SK-PN-DW Ewing cell lines to assess the role of CCK in cell proliferation and tumor growth. Results: Microarray analysis revealed that CCK was up-regulated by EWS/FLI1 in HeLa cells. CCK was overexpressed in Ewing tumors as compared with other pediatric malignancies such as rhabdomyosarcoma and neuroblastoma, with levels close to those detected in normal tissues expressing the highest levels of CCK. Furthermore, EWS/FLI1 knockdown in A673 and SK-PN-DW Ewing cells using two different doxycycline-inducible EWS/FLI1-specific shRNA vectors down-regulated CCK mRNA expression and diminished the levels of secreted CCK, showing that CCK is a EWS/FLI1 specific target gene in Ewing cells. A doxycycline-inducible CCK-specific shRNA vector successfully down-regulated CCK expression, reduced the levels of secreted CCK in Ewing cell lines, and inhibited cell growth and proliferation in vitro and in vivo. Finally, we show that Ewing cell lines and tumors express CCK receptors and that the growth inhibition produced by CCK silencing can be rescued by culturing the cells with medium containing CCK. Conclusions: Our data support the hypothesis that CCK acts as an autocrine growth factor stimulating the proliferation of Ewing cells and suggest that therapies targeting CCK could be promising in the treatment of Ewing tumors.Ewing family of tumors is a group of highly malignant tumors arising mainly in bone that most often affects children and young adults in the first two decades of life. Despite the use of multimodal therapy (chemotherapy, radiation therapy, and surgery), the long-term disease-free survival rate of Ewing tumor patients is still disappointingly low, particularly in the high-risk groups with metastasis at diagnosis or unfavorable localization (reviewed in ref. 1). The lack of new effective drugs in the treatment of Ewing tumors, together with the side effects that high-dose regimens have on young patients who have a long life expectancy, supports the need for innovative therapeutic strategies that should mainly include targeted therapies against molecules critical for the pathogenesis and progression of these tumors.The molecular hallmark of Ewing family of tumors is the presence of balanced chromosomal translocations leading to the formation of chimeric transcription factors. These aberrant...

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Section: Discussionsupporting

confidence: 79%

Cholecystokinin Down-Regulation by RNA Interference Impairs Ewing Tumor Growth

Carrillo

García-Aragoncillo

Azorín

et al. 2007

Clinical Cancer Research

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“…Expression of neural structural genes, such as neuronal pentraxin receptor, synaptotagmin I, and MAPT, is also highly induced. CCK is a neuropeptide and is highly expressed in EFT (46). This is almost unique among tumor cells and served as evidence that EFT originated from parasympathetic progenitors.…”

Section: Ews-fl11 Targets Neural Crest Development Genesmentioning

confidence: 99%

EWS-FLI1 Fusion Protein Up-regulates Critical Genes in Neural Crest Development and Is Responsible for the Observed Phenotype of Ewing's Family of Tumors

et al. 2005

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Tumor-specific translocations are common in tumors of mesenchymal origin. Whether the translocation determines the phenotype, or vice versa, is debatable. Ewing's family tumors (EFT) are consistently associated with an EWS-FLI1 translocation and a primitive neural phenotype. Histogenesis and classification are therefore uncertain. To test whether EWS-FLI1 fusion gene expression is responsible for the primitive neuroectodermal phenotype of EFT, we established a tetracycline-inducible EWS-FLI1 expression system in a rhabdomyosarcoma cell line RD. Cell morphology changed after EWS-FLI1 expression, resembling cultured EFT cells. Xenografts showed typical EFT features, distinct from tumors formed by parental RD. Neuron-specific microtubule gene MAPT, parasympathetic marker cholecystokinin, and epithelial marker keratin 18 were up-regulated. Conversely, myogenesis was diminished. Comparison of the up-regulated genes in RD-EF with the Ewing's signature genes identified important EWS-FLI1 downstream genes, many involved in neural crest differentiation. These results were validated by real-time reverse transcription-PCR analysis and RNA interference technology using small interfering RNA against EWS-FLI1 breakpoint. The present study shows that the neural phenotype of Ewing's tumors is attributable to the EWS-FLI1 expression and the resultant phenotype resembles developing neural crest. Such tumors have a limited neural phenotype regardless of tissue of origin. These findings challenge traditional views of histogenesis and tumor origin. (Cancer Res 2005; 65(11): 4633-44)

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“…In addition to newly identified genes with high expression in EFTs, we found several genes known to be highly expressed in EFTs, e.g., cholecystokinin (CCK; ref. 25 (29). In addition, many genes were not detected in EFTs or were underrepresented in EFTs compared with BM.…”

Section: Comparison Of Gene Expression Profiles Of Efts and Normalmentioning

confidence: 99%

DNA Microarrays Reveal Relationship of Ewing Family Tumors to Both Endothelial and Fetal Neural Crest-Derived Cells and Define Novel Targets

et al. 2004

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Ewing family tumors (EFTs) are small round blue cell tumors that show features of neuroectodermal differentiation. However, the histogenetic origin of EFTs is still a matter of debate. We used high-density DNA microarrays for the identification of EFT-specific gene expression profiles in comparison with normal tissues of diverse origin. We identified 37 genes that are up-regulated in EFTs compared with normal tissues and validated expression of these genes in EFTs by both conventional and quantitative reverse transcription-polymerase chain reaction. The expression pattern of EFT-associated genes in normal tissues indicated a high similarity between EFTs and fetal and neuronal as well as endothelial tissues and supports the concept that a primitive neural crest-derived progenitor at the transition to mesenchymal and endothelial differentiation is transformed in EFTs. EFT-associated genes could be used for molecular discrimination between EFTs and other small round blue cell tumors and clearly identified a cell line (SK-N-MC) that was initially established as neuroblastoma as being an EFT. Ectopic expression of the EFT-specific EWS-FLI1 fusion protein in human embryonic kidney (HEK293) cells was not sufficient to induce the complete EFT-specific gene expression signature, suggesting that the EFT-specific gene expression profile is not just a consequence of EWS-FLI1 expression but depends on the histogenetic background of the EFT stem cell.

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Expression of the cholecystokinin gene in pediatric tumors.

Cited by 16 publications

References 34 publications

Cholecystokinin Down-Regulation by RNA Interference Impairs Ewing Tumor Growth

Cholecystokinin Down-Regulation by RNA Interference Impairs Ewing Tumor Growth

EWS-FLI1 Fusion Protein Up-regulates Critical Genes in Neural Crest Development and Is Responsible for the Observed Phenotype of Ewing's Family of Tumors

DNA Microarrays Reveal Relationship of Ewing Family Tumors to Both Endothelial and Fetal Neural Crest-Derived Cells and Define Novel Targets

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