HO-1 modulates the expression of genes involved in inflammation, angiogenesis, proliferation, apoptosis and cell adhesion in human lung cancer cells.
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ABSTRACT: The enzymatic activity of HO-1 results in decreased oxidative stress, attenuated inflammatory response, and very often in a lower rate of apoptosis. This is due to removal of heme, a potent prooxidant and proinflammatory agent, but mainly because of generation of biologically active products such as CO and bilirubin. In order to find the correlation between the HO-1 level and the expression of different genes of interest we have utilized human lung cancer cell line NCI-H292 stably overexpressing HO-1. Additionally we have checked if HO-1 can modulate genes expression in NCI-H292 cells treated with TNF. The effect of HO-1 overexpression on transcriptome was assessed by microarray analysis. We have observed that the increase in HO-1 level may affect the expression of different genes involved in cytokine synthesis, angiogenesis, apoptosis, proliferation and cell adhesion. Additionally, HO-1 may interact with the TNF treatment and influence the expression of some genes like IL-1, IL-6, IL-8, FAS and NF-kB.
Project description:The enzymatic activity of HO-1 results in decreased oxidative stress, attenuated inflammatory response, and very often in a lower rate of apoptosis. This is due to removal of heme, a potent prooxidant and proinflammatory agent, but mainly because of generation of biologically active products such as CO and bilirubin. In order to find the correlation between the HO-1 level and the expression of different genes of interest we have utilized human lung cancer cell line NCI-H292 stably overexpressing HO-1. Additionally we have checked if HO-1 can modulate genes expression in NCI-H292 cells treated with TNF. The effect of HO-1 overexpression on transcriptome was assessed by microarray analysis. We have observed that the increase in HO-1 level may affect the expression of different genes involved in cytokine synthesis, angiogenesis, apoptosis, proliferation and cell adhesion. Additionally, HO-1 may interact with the TNF treatment and influence the expression of some genes like IL-1, IL-6, IL-8, FAS and NF-kB. NCI-H292 cells, control (pcDNA) and stably overexpressig HO-1 (pcHO1) were treated for 24 h with TNF (30 ng/ul), then the cells were lysed and RNA was collected. The whole human gene expression profile was assessed with Agilent microarray analysis. Three independent experiments were performed at each time using different batch of 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.
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.
Project description:Nrf2 antioxidant signaling is involved in liver protection, but this generalization overlooks conflicting studies indicating that Nrf2 effects are not necessarily hepatoprotective. The role of Nrf2/HO-1 in cholestatic liver injury (CLI) remains poorly defined. Here, we report that Nrf2/HO-1 activation exacerbates liver injury rather than exerts a protective effect in CLI. Inhibiting HO-1 or ameliorating bilirubin transport alleviates liver injury in CLI models. Nrf2 knockout confers hepatoprotection in CLI mice, whereas in non-CLI mice, Nrf2 knockout aggravates liver damage. In the CLI setting, oxidative stress activates Nrf2/HO-1, leads to bilirubin accumulation, and impairs mitochondrial function. High levels of bilirubin reciprocally upregulate the activation of Nrf2 and HO-1, while antioxidant and mitochondria-targeted SOD2 overexpression attenuate the toxicity of bilirubin. Additionally, the expression of Nrf2 and HO-1 is significantly elevated in serum of patients with CLI. These results reveal an unrecognized function of Nrf2 signaling in exacerbating liver injury in cholestatic disease.
Project description:Sepsis-associated encephalopathy (SAE) affects up to 70% ofSepsis-associated encephalopathy (SAE) affects up to 70% of patients in intensive care units with severe systemic infection. However, the exact pathological mechanisms behind SAE remain unclear. In this study, we aimed to investigate the protective effects of flavonoid components extracted from CCL seeds on SAE animals and evaluate the transcriptomic alterations in the hippocampus using RNA sequencing. Our results showed that CCL seed extract improved learning and memory abilities and structural integrity of the blood-brain barrier in CLP-induced SAE animal models. RNA sequencing revealed that CCL treatment reversed SAE-induced transcriptomic alterations in the hippocampus. Additionally, CCL significantly reduced inflammation (TNF-α, IL-2, and IL-6) and oxidative stress (MDA and SOD activity), as well as inhibited neuron apoptosis in brain tissues. Furthermore, CCL restored the PI3K/AKT signaling pathway, leading to Nrf2 nuclear translocation and HO-1 expression both in vitro and in vivo. The PI3K inhibitor LY294002 blocked CCL's anti-apoptotic, anti-inflammatory, and anti-oxidative effects, demonstrating that CCL's bioactivities are dependent on the PI3K/AKT signaling pathway. In conclusion, CCL exhibits significant neuroprotective properties and may be a promising candidate for further clinical trials in SAE treatment.
Project description:To investigate the gene expression of lung epithelial cells effected by Trichomonas tenax, we chose NCI-H292 lung epithelial cells and cocultured with Trichomonas tenax.
Project description:Heme Oxygenase-1 (HO-1) is expressed in many cancers and influences the growth, survivall and metastasis of tumors, however, the molecular mechanisms remains largely unknown. To identify a common mechanism of action of HO-1 in cancer, we studied the global effect of HO-1 on the transcriptome of multiple tumors. Genome-wide expression profiling of HO-1 expressing versus HO-1 silenced cancer cells and a further data mining analysis of the preexisting expression database of 190 human tumors of 14 cancer types led us to identify 14 genes, the expression of which correlated firmly and universally with that of HO-1 (P < 0.001). These genes included regulators of cell plasticity and extracellular matrix remodeling (MMP2, ADAM8, TGFβ1, BGN, COL21A1, PXDN), signaling (CRIP2, MICB), amino acid transport and glycosylation (SLC7A1 and ST3GAL2), estrogen and phospholipid biosynthesis (AGPAT2 and HSD17B1), protein stabilization (IFI30) and phosphorylation (ALPPL2). PXDN, one of the genes being co-expressed with HO-1, was selected for further analysis. Immunofluorescence and western blotting confirmed positive correlation of PXDN with HO-1 levels in BeWo cancer cells as well as co-localization in invasive extravillous trophoblast cells of first trimester placenta. Loss of HO-1 in BeWo cells correlated with reduced cell adhesion to Collagen type I, Fibronectin and Laminin. The adhesion-promoting effects of HO-1 were dependent on PXDN expression, as loss of PXDN in HO-1 expressing BeWo cells led to reduced cell attachment to Laminin and Fibronectin coated wells.
Project description:Heme Oxygenase-1 (HO-1) is expressed in many cancers and influences the growth, survivall and metastasis of tumors, however, the molecular mechanisms remains largely unknown. To identify a common mechanism of action of HO-1 in cancer, we studied the global effect of HO-1 on the transcriptome of multiple tumors. Genome-wide expression profiling of HO-1 expressing versus HO-1 silenced cancer cells and a further data mining analysis of the preexisting expression database of 190 human tumors of 14 cancer types led us to identify 14 genes, the expression of which correlated firmly and universally with that of HO-1 (P < 0.001). These genes included regulators of cell plasticity and extracellular matrix remodeling (MMP2, ADAM8, TGFβ1, BGN, COL21A1, PXDN), signaling (CRIP2, MICB), amino acid transport and glycosylation (SLC7A1 and ST3GAL2), estrogen and phospholipid biosynthesis (AGPAT2 and HSD17B1), protein stabilization (IFI30) and phosphorylation (ALPPL2). PXDN, one of the genes being co-expressed with HO-1, was selected for further analysis. Immunofluorescence and western blotting confirmed positive correlation of PXDN with HO-1 levels in BeWo cancer cells as well as co-localization in invasive extravillous trophoblast cells of first trimester placenta. Loss of HO-1 in BeWo cells correlated with reduced cell adhesion to Collagen type I, Fibronectin and Laminin. The adhesion-promoting effects of HO-1 were dependent on PXDN expression, as loss of PXDN in HO-1 expressing BeWo cells led to reduced cell attachment to Laminin and Fibronectin coated wells. We used gene expression profiling to determine the genome-wide effect of HO-1 on the transcriptome of BeWo trophoblast cells. We specifically selected BeWo cells for our studies because these cells express HO-1 naturally. We knocked down endogenous HO-1 in BeWo cells using retroviral transduction with a micro-RNA adapted retroviral vector targeting human HO-1 sequence. RNA isolated from control (LMP) or miHO1 infected (miHO-1) cells was labeled and hybridized to human genome-wide gene level 1.0 ST arrays
Project description:To clarify the role of Caspase-4 in non-small cell lung cancer cells, Caspase-4 overexpressing and knockout cells in NCI-H292 cells were established and each was analyzed by RNAseq.