Unknown,Transcriptomics,Genomics,Proteomics

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RMECs: hypoxia-induced group vs. paired untreated group

ABSTRACT: Objective To explore the molecular mechanisms of retinopathy of prematurity (ROP) at whole genome scale using cultivated human fetal retinal microvascular endothelial cells (RMECs) under hypoxic condition. Study design Aborted fetuses with mean gestational age of 20-28 weeks at Guangdong Women and ChildrenM-bM-^@M-^Y Hospital were enrolled, 3 males and 2 females, with mean weight 610-2205 g. Primary RMECs were isolated by mechanical morcel and trypsin-collagenase digestion of eyeballs. Cultivated cells were treated with or without 150 M-NM-2.0 and q-value <0.05. Gene Ontology was employed for functional enrichment analysis. Results There were 326 DEGS between the hypoxia-induced group and untreated group. Of these genes, 198 were up-regulated in hypoxic RMECs, while other 128 hits were down-regulated. More importantly, altered expression genes in iron ion homeostasis pathway were highly enriched under hypoxic condition. Quantitative RT-PCR (qRT-PCR) was performed to validate the results obtained from microarray analysis. Conclusions Dysregulation of genes involved in iron homeostasis mediating oxidative damage may contribute to the ROP pathogenesis. Primary RMECs isolated from five subjects (3 males, 2 females) with mean weight 610-2205 g. CoCl2 induced hypoxia in primary RMECs vs. cultured cells without CoCl2 treatment. Biological replicates: five treated RMECs and paired untreated control. Dye-swap assays were performed for each sample.

ORGANISM(S): Homo sapiens

SUBMITTER: Liang Zhang

PROVIDER: E-GEOD-59462 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Using the orthotopic breast cancer xenograft model of basal-like breast cancer MDA-MB-231 line, we have found that the expression of miRNAs encoded by MIR17HG was significantly decreased in cells isolated from spontaneous lung metastases compared to cells from primary tumors grown in orthotopic sites. We investigated the role of a MIR17HG family member, miR-18a, in primary tumor growth and pulmonary metastasis from the orthotopic site. We demonstrated that enforced expression of exogenous miR-18a, significantly limited continuous growth of primary tumors in mammary gland fat pads and reduced spontaneous lung metastasis. Further investigation on the mechanism of miR-18a action led to the finding that the expression of HIF1A, a key regulator of tumor metastasis, was regulated by miR-18a. Enforced miR-18a expression significantly decreased HIF1A expression at both mRNA and protein levels, resulting in altered transcriptional response and decreased survival of cells in response to Cobalt(II) chloride (CoCl2), a hypoxia-mimicking agent. Conversely, miR-18a knockdown significantly increased HIF1A expression levels and enhanced cell survival in response to CoCl2. Analysis of expression data of human breast tumor tissues showed that miR-18a expression is inversely correlated with HIF1A expression in basal-like breast tumors, supporting a role of miR-18a in restricting HIF1A expression in this subtype of breast cancer. In addition, we demonstrated that hypoxia inhibits miR-18a expression, likely through MYC inactivation. Furthermore, gene expression and functional analysis revealed that miR-18a also plays a role in regulating cell adhesion, migration and invasion. Taken together, this study provides evidence for a novel role of miR-18a to inhibit breast cancer metastasis. Our results suggest that miR-18a downregulation might provide tumor cells survival/growth advantage under hypoxic pressure in basal-like breast cancer. A lung metastatic subline, designated as MB231RN-LM, was derived from MDA-MB-231 breast cancer cells through in vivo selection. The MB231RN-LM cells were stably transfected with has-miR-18a or control vector and treated with 200uM Cobalt(II) chloride (CoCl2, a hypoxia-mimicking agent) for 4 hr. A total of 8 samples were subjected to microarray analysis, with two biological repeats for each experiment condition.

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RMECs: hypoxia-induced group vs. paired untreated group

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