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The effect of HO-1 overexpression on microRNA transcriptome in NCI-H292 lung cancer cells

ABSTRACT: The aim was to determine the effect of heme oxygenase-1 (HO-1) overexpression on microRNA transcriptome in human non-small cell lung carcinoma cell line (NCI-H292). Since the cells of different HO-1 genotypes were used (cells are after retroviral transduction with empty vector with normal level of HO-1 or retroviral transduction with vector harboring HO-1), it is possible get the comprehensive answer which microRNAs are regulated by HO-1. Confluent NCI-H292 cells that contain EV-ctrl or overexpressing HO-1. Pool of reference cells are cells untreated and treated with 10 ng/ml TNF-α for 6 hours before RNA isolation. The samples are biological triplicates - three independent experiments were performed at different time points for all cell lines. Total number of the presented samples is 6.

ORGANISM(S): Homo sapiens

SUBMITTER: Skrzypek K

PROVIDER: S-ECPF-GEOD-37779 | biostudies-other | 2013 Sep

REPOSITORIES: biostudies-other

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The effect of HO-1 overexpression on microRNA transcriptome in NCI-H292 lung cancer cells

Project description:The aim was to determine the effect of heme oxygenase-1 (HO-1) overexpression on microRNA transcriptome in human non-small cell lung carcinoma cell line (NCI-H292). Since the cells of different HO-1 genotypes were used (cells are after retroviral transduction with empty vector with normal level of HO-1 or retroviral transduction with vector harboring HO-1), it is possible get the comprehensive answer which microRNAs are regulated by HO-1. Confluent NCI-H292 cells that contain EV-ctrl or overexpressing HO-1. Pool of reference cells are cells untreated and treated with 10 ng/ml TNF-M-NM-1 for 6 hours before RNA isolation. The samples are biological triplicates - three independent experiments were performed at different time points for all cell lines. Total number of the presented samples is 6.

2013-04-27 | E-GEOD-37779 | biostudies-arrayexpress

The effect of HO-1 overexpression on microRNA transcriptome in NCI-H292 lung cancer cells

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mRNA and microRNA expression profiles of radioresistant NCI-H520 non-small cell lung cancer cells.

Project description:To elucidate the mechanism of radioresistance in non-small cell lung cancer (NSCLC) cells and to identify key molecules conferring radioresistance, the radioresistant subclone NCI-H520/R, derived from the NCI-H520 NSCLC cell line, was established with eight rounds of sublethal irradiation. The radioresistant features were subsequently assessed using a clonogenic assay, analysis of apoptosis and an MTT assay, the gene expression levels were examined using an Agilent Whole Human Genome 4 x 44 k Oligo microarray and Agilent Human miRCURY(™) LNA array, and confirmed by reverse transcription-quantitative polymerase chain reaction. Pathway analysis and Gene Ontology (GO) analysis were performed to determine the biological functions of the subset of differentially expressed genes. miRNA-mRNA correlation analysis between the expression levels of each miRNA and all its predicted target genes was performed to further understand the radioresistance in the NCI-H520 cells. Following eight rounds of sublethal irradiation, a total of 2,862 mRNAs were significantly differentially expressed in the NCI-H520/R cells, including 893 upregulated genes and 1,969 downregulated genes. A total of 162 upregulated miRNAs and 274 downregulated miRNAs were significantly deregulated in the NCI-H520/R cells. Multiple core regulatory processes and signaling pathways were identified as being of likely relevance to radioresistance in NCI-H520/R cells, including the mitogen-activated protein kinase signaling pathway and neurotrophin signaling pathway. The expression of genes associated with radioresistance reflects the complex biological processes involved in clinical cancer cell eradication and requires further investigation for future enhancement of therapy.

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MicroRNA response to Snail overexpression in lung cancer cell lines

Project description:Analysis of four lung cancer cell lines transfected with a vector expressing the transcriptional repressor Snail versus a vector control. Aberrant Snail expression is known to induce an EMT program in lung cancers. Four lung cancer cell lines (H292, H358, H441, H1437) with stable overexpression of the human SNAI1 gene and vector expressing controls were collected at equal confluency with the miRNeasy Mini kit (Qiagen). One microgram of total RNA was labeled using miRCURY LNA™ microRNA Array Power Labeling kit by the UCLA Clinical Microarray Core. The labeled miRNAs were hybridized to Exiqon miRCURY LNA microRNA Array-6th Generation according to the manufacturer’s instructions. This array includes 927/648/351 human/mouse/rat miRNAs as well as 438 miRPlus miRNAs. The miRNA array slides were scanned using Axon GenePix 4000B scanner (Axon Instruments, Foster City, CA) and processed by using the GenePix Pro 6.0 software (Axon Instruments). The raw data were normalized by using a combination of housekeeping miRNA, spike-in miRNA and invariant endogenous miRNAs.

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Effect of cigarette smoke on microRNA expression in H292 lung cancer cells

Project description:Cigarette smoke has been associated with the development of various lung diseases including cancer. Dysregulation of miRNAs is known to affect protein expression which leads to diverse functional consequences. Investigating miRNA and protein expression in response to cigarette smoke exposure can lead to the identification of potential therapeutic and chemopreventive targets. We employed a SILAC-based quantitative proteomic analysis to identify proteins differentially expressed in response to cigarette smoke in H292 lung cancer cells. LC-MS/MS analysis led to the identification of 3,959 proteins, of which, 303 proteins were overexpressed and 112 proteins were found to be downregulated in cigarette smoke-treated H292 cells.

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Effect of pioglitazone treatment on gene expression in NCI-H2347 lung cancer cells: time course experiment

Project description:Microarray studies was performed to analyze gene expression changes in NCI-H2347 cells after treatment with 50 µM pioglitazone for 12hr, 24hr and 48hrs. RNA samples, containing 3 replicates for each sample, were extracted from NCI-H2347 cells after treatment with pioglitazone or vhicle (DMSO) for 12hr, 24hr and 48hrs. After biotin labeling, samples were hybridized on the Illumina Human HT12v4.0 Expression Beadchip.

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Transcriptome sequencing of human lung cancer cell line NCI-H838 treated with LINC00472 overexpression

Project description:To evaluate the changes in the expression of genes in lung cancer cells due to LINC00472, we performed transcriptome sequencing of LINC00472 overexpressing cells and control cells. First, raw reads were obtained through sequencing of control cells not treated with LINC00472 and cells treated with overexpression of LINC00472. Thereafter, genes differentially expressed in the two groups were selected through pretreatment, mapping to the reference genome, trascripts assaembly, and transcript quantification. The sequencing result provides quantitative and qualitative information of total RNAs whose expression is changed by LINC00472. This information will facilitate analysis of the signaling network of LINC00472 in lung cancer cells.

2021-06-30 | GSE164230 | GEO

SOX2 overexpression effect on human lung squamous cells

Project description:We have identified SOX2 as a new oncogene and a likely driver of recurrent 3q26.3 amplifications in lung Squamous Cell Carcinoma. SOX2 is a crucial transcription factor implicated in Embryonic and Neural Stem Cells, that we found widely activatd in human lung SCC. This part of the study aimed at analyzing the transcriptomic consequences of SOX2 overexpression in a simple in vitro model (human lung squamous immortalized cells). Human lung squamous BEAS-2B cells were transduced for Control or SOX2 expression, and their transcriptomes compared using Affymetrix arrays.

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Co-overexpression of Met and hepatocyte growth factor promotes systemic metastasis in NCI-H460 non-small cell lung carcinoma cells.

Project description:Complete resection of early-stage non-small cell lung cancer (NSCLC) is potentially curative, yet approximately 50% of patients are at risk for developing metastatic recurrence. Met, the receptor for hepatocyte growth factor (HGF) is a receptor tyrosine kinase with demonstrated roles in regulating cellular proliferation, motility, morphogenesis, and apoptosis. Met receptor and its ligand, HGF, are commonly overexpressed in NSCLC, and their overexpression has been associated with poor prognosis, which could potentially involve a paracrine and/or autocrine activation loop. However, there is as yet no direct evidence that HGF-Met signaling directly promotes metastasis in NSCLC cells. Using retroviral transduction, we overexpressed the human c-met and hgf complementary DNA, alone or in combination in the NCI-H460 human large cell carcinoma cell line. The HGF/Met co-overexpressing (H460-HGF/Met) cells demonstrated enhanced tumorigenicity in xenograft SCID mice. When these cells are implanted orthotopically into the lungs of nude rats, only the H460-HGF/Met cells showed higher spontaneous metastases to distant organs including bone, brain, and kidney. These results provide evidence that autocrine overactivation of the Met- HGF loop enhances systemic metastases in NSCLC. Targeted interference of this loop may potentially be an effective adjuvant therapy to improve survival of early-stage NSCLC patients.

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Luteolin exerts an anticancer effect on NCI-H460 human non-small cell lung cancer cells through the induction of Sirt1-mediated apoptosis.

Project description:Luteolin is a falconoid compound, which exhibits anticancer properties, however, its contribution to Sirt1-mediated apoptosis in human non-small cell lung cancer remains to be elucidated. The present study confirmed that the anticancer effect of luteolin on NCI‑H460 cells was through Sirt1‑mediated apoptosis. The NCI‑H460 cells were treated with different concentrations of luteolin, and a 3‑(4,5‑dimethyl‑2‑thiazolyl)‑2,5‑diphnyl‑2H‑tetrazolium bromide assay, cell cycle analysis and annexin‑V/fluorescein isothiocyanate and propidium double staining were performed to assess the apoptotic effect of luteolin. Wound healing and Transwell assays were performed to confirm the inhibition of NCI‑H460 cell migration. The protein levels of Sirt1 were knocked down in the NCI‑H460 cells using a lentivirus to further investigate the role of this protein, and the expression levels of the apoptotic associated proteins, Bad, Bcl‑2, Bax, caspase‑3 and Sirt1, were measured using western blotting. The results of the present study demonstrated that luteolin exerted an anticancer effect against NCI‑H460 cells through Sirt1‑mediated apoptosis and the inhibition of cell migration.

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