Unknown,Transcriptomics,Genomics,Proteomics

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Supernatants derived from chemotherapy-treated tumours can modify angiogenesis.

ABSTRACT: The human lung cancer cell line A549 was cultured in media alone, with cyclophosphamide (CPM) or oxaliplatin (OXP) for 72 hours. The exhausted culture media were aspirated and the microvesicular fraction was harvested by sequential ultracentrifugation. RNA was then isolated from microvesicles by Trizol. Total RNA was labelled and hybridised to Illumina Human HT-12v3 arrays.

ORGANISM(S): Homo sapiens

SUBMITTER: Wai Liu

PROVIDER: E-TABM-1201 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Supernatants derived from chemotherapy-treated cancer cell lines can modify angiogenesis.

Liu W M WM Dennis J L JL Gravett A M AM Chanthirakumar C C Kaminska E E Coulton G G Fowler D W DW Bodman-Smith M M Dalgleish A G AG

British journal of cancer 20120131 5

<h4>Background</h4>There is evidence that tumours produce substances such as cytokines and microvesicular bodies bearing bioactive molecules, which support the carcinogenic process. Furthermore, chemotherapy has also been shown to modify these exudates and in doing so, neutralise their tumourigenic influence.<h4>Methods</h4>In the current study, we have investigated the effect of chemotherapy agents on modifying the cytokine profile and microvesicular cargo of supernatants derived from cancer ce ...[more]

PMID: 22294186

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Project description:We previously characterized zinc finger protein gene HZF1 (ZNF16) and demonstrated its important roles in erythroid and megakaryocytic differentiation of K562 cells by loss-function assay. However its effect in erythroid and megakaryocytic differentiation of hematopoietic stem/progenitor cells (HSPCs) and the mechanisms by which it functions have not been understood. In this study, we detected up-regulation of ZNF16 during erythroid and megakaryocytic differentiation of K562 cells and normal CD34+ HSPCs, and demonstrated that ZNF16 promotes erythroid and megakaryocytic differentiation by gain-of-function and loss-of-function experiments. Gene expression profiling by mRNA array and PCR validation in the K562 transforments with ZNF16 over-expression suggested that cell division cycle-associated 7-like gene (JPO2) and v-kit Hardy-Zuckerman 4 feline sarcoma viral oncogene homolog gene (c-KIT) were among the genes regulated possibly by ZNF16. Luciferase reporter assay and Chromatin Immunoprecipitation demonstrated that ZNF16 binds to JPO2 and c-KIT promoters and inhibits their expression in K562 cells. A significant decrease of JPO2 and c-KIT levels was observed during erythroid and megakaryocytic differentiation of K562 and CD34+ cells. The knockdown of either JPO2 or c-KIT partially rescued the differentiation inhibition caused by ZNF16 knockdown. We also found that ZNF16 inhibits c-KIT/c-Raf/MEK/ERK/c-Jun/HEY1 signal pathways, which finally up-regulated expression of GATA1, a central regulator of erthroid and megakaryocyte differentiation. By lentivirus-mediated gene transfer, we demonstrated that enforced expression and knockdown of ZNF16 in HSPCs down-regulated and up-regulated expression of its targets respectively. Our data collectively demonstrate that ZNF16 promotes erythropoiesis and megakaryocytopoiesis via its regulation on JPO2 and c-KIT. 4 samples are analyzed, H4-1 and H4-2 are two stable K562 transductants with ZNF16 over-expression, PC1 and PC2 are stable control K562 transductants. Stable K562 transductants were obtained for RNA extraction and hybridization on an Illumina HumanHT-12 V4.0 expression beadchipe xpression Array platform.

Dataset Information

Supernatants derived from chemotherapy-treated tumours can modify angiogenesis.

Publications

Supernatants derived from chemotherapy-treated cancer cell lines can modify angiogenesis.

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