Treatment Of A549 Cells Research Articles

Analysis of Polyphyllin II alters microRNA expression profile in lung cancer A549 cells

Non-small cell lung cancer (NSCLC) is a malignant tumor that threatens the whole world, previous studies found that Polyphyllin II (PPII) can suppress NSCLC cells growth, exhibits significant antitumor activity. MicroRNAs (miRNAs) can regulate the expression of other genes and play a crucial role in the prevention and development of cancer. However, the miRNA portrait of ginsenoside PPII-treated NSCLC A549 cells has not yet been studied. In this work, miRNA-seq analysis was used to determine the changes in miRNA expression profile of NSCLC A549 cells PPII-treaded. In addition, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis were performed on differentially expressed miRNA target genes. Then, predicted the target genes of miRNA and performed functional enrichment analysis on them. We identified 58 up-regulated and 24 down-regulated miRNAs displaying changes their expression in PPII treated A549 cells were greater than 2-fold. In addition, the expression levels of miRNAs were validated by real-time PCR. Therefore, this work has a certain promoting effect on the study of the anti-cancer mechanism of PPII in lung cells.

LncRNA SNHG1 knockdown attenuates lipopolysaccharide-induced acute lung injury via regulating miR199a-3p/ROCK2 axis.

Acute lung injury (ALI) is a significant health condition with notable rates of morbidity and mortality globally. Long non-coding ribose nucleic acids (lncRNAs) play vital roles in mitigating various inflammation-related diseases, including ALI. The study aimed to investigate the functional role and molecular mechanisms of lncRNA SNHG1 on ALI in lipopolysaccharide (LPS)-treated A549 cells and in LPS-induced ALI mice. The expression of SNHG1 was initially examined in LPS-treated A549 cells. We further demonstrated the critical function of SNHG1 through various cellular assessments following SNHG1 knockdown, including cell counting kit (CCK)-8 assay, flow cytometry analysis, as well as enzyme-linked immunosorbent assay (ELISA). Reducing SNHG1 levels hindered the negative effects of LPS on cell viability, apoptosis, and inflammation. Moreover, SNHG1 acted as a negative regulator for miR-199a-3p, which targeted downstream ROCK2. Depletion of miR-199a-3p or enhanced expression of ROCK2 abolished the protective effects of SNHG1 knockdown on LPS-induced apoptosis and inflammation. Consistently, silencing SNHG1 alleviated LPS-induced lung injury in mice, demonstrating its potential therapeutic benefits in managing ALI. Overall, this study sheds light on the role of SNHG1 in modulating inflammation and apoptosis in ALI through the miR-199a-3p/ROCK2 pathway, offering new insights for the treatment of this condition.

Polyamine Catabolism Revisited: Acetylpolyamine Oxidase Plays a Minor Role due to Low Expression.

Biogenic polyamines are ubiquitous compounds. Dysregulation of their metabolism is associated with the development of various pathologies, including cancer, hyperproliferative diseases, and infections. The canonical pathway of polyamine catabolism includes acetylation of spermine and spermidine and subsequent acetylpolyamine oxidase (PAOX)-mediated oxidation of acetylpolyamines (back-conversion) or their direct efflux from the cell. PAOX is considered to catalyze a non-rate-limiting catabolic step. Here, we show that PAOX transcription levels are extremely low in various tumor- and non-tumor cell lines and, in most cases, do not change in response to altered polyamine metabolism. Its enzymatic activity is undetectable in the majority of cell lines except for neuroblastoma and low passage glioblastoma cell lines. Treatment of A549 cells with N1,N11-diethylnorspermine leads to PAOX induction, but its contribution to polyamine catabolism remains moderate. We also describe two alternative enzyme isoforms and show that isoform 4 has diminished oxidase activity and isoform 2 is inactive. PAOX overexpression correlates with the resistance of cancer cells to genotoxic antitumor drugs, indicating that PAOX may be a useful therapeutic target. Finally, PAOX is dispensable for the replication of various viruses. These data suggest that a decrease in polyamine levels is achieved predominantly by the secretion of acetylated spermine and spermidine rather than by back-conversion.

Green synthesis of copper oxide nanoparticles using genus Inula and evaluation of biological therapeutics and environmental applications

Abstract In this research, we produced copper oxide nanoparticles (CuO NPs) using extracts from the entire above-ground portion of plants of genus Inula (Inula graveolens). The synthesis of CuO NPs was verified through various physicochemical analytical methods, including UV–visible, Fourier transform infrared, and transmission electron microscopy. The CuO NPs were found to be around 20 nm in size and spherical in shape. Subsequently, the synthesized compounds were evaluated for their anticancer properties. After treating A549 cells with CuO NPs at concentrations of 15 and 30 μg, we examined their cytotoxicity, lipid peroxidation activity (malondialdehyde level), and antioxidant activity (catalase, superoxide dismutase, and glutathione levels). Additionally, we analyzed the expression of apoptotic marker genes (p53, caspase-3, and caspase-9), cytokine levels (IL-6 and TNF-α), and DNA fragmentation. Our findings demonstrated that CuO NPs enhanced the expression of apoptotic genes, suggesting that phytochemical-derived NPs from Inula extracts induce apoptosis by upregulating tumor suppressor genes and downregulating oncogenes in A549 cells. Furthermore, CuO NPs exhibited higher susceptibility toward B. subtilis and S. aureus compared to ampicillin. Using the response surface methodology, we determined that CuO NPs are effective adsorbents for removing Pb2+ ions from aqueous solutions, making them promising for environmental applications. Overall, our results indicate that CuO NPs have potential as antimicrobial, antioxidant, and anticancer agents and as efficient adsorbents.

Apoptotic efficiency of Dicoma anomala biosynthesized silver nanoparticles against A549 lung cancer cells

Lung cancer is one of the common forms of cancer that affects both men and women and is regarded as the leading cause of cancer related deaths. It is characterized by unregulated cell division of altered cells within the lung tissues. Green nanotechnology is a promising therapeutic option that is adopted in cancer research. Dicoma anomala (D. anomala) is one of the commonly used African medicinal plant in the treatment of different medical conditions including cancer. In the present study, silver nanoparticles (AgNPs) were synthesized using D. anomala MeOH root extract. We evaluated the anticancer efficacy of the synthesized AgNPs as an individual treatment as well as in combination with pheophorbide a (PPBa) mediated photodynamic therapy (PDT) in vitro. UV–VIS spectroscopy, high-resolution transmission electron microscopy (HR-TEM), Scanning electron microscopy (SEM) and energy dispersive X-ray spectroscopy (EDS) was used to confirm the formation of D.A AgNPs. Post 24 h treatment, A549 cells were evaluated for ATP proliferation, morphological changes supported by LIVE/DEAD assay, and caspase activities. All experiments were repeated four times (n=4), with findings being analysed using SPSS statistical software version 27 set at 0.95 confidence interval. The results from the present study revealed a dose-dependent decrease in cell proliferation in both individual and combination therapy of PPBa mediated PDT and D.A AgNPs on A549 lung cancer cells with significant morphological changes. Additionally, LIVE/DEAD assay displayed a significant increase in the number of dead cell population in individual treatments (i.e., IC50’s treated A549 cells) as well as in combination therapy. In conclusion, the findings from this study demonstrated the anticancer efficacy of green synthesized AgNPs as a mono-therapeutic drug as well as in combination with a chlorophyll derivative PPBa in PDT. Taken together, the findings highlight the therapeutic potential of green nanotechnology in medicine.

Effect of LRRC15 on autophagy in A549 cells.

Idiopathic pulmonary fibrosis (IPF) is a progressive, chronic, and irreversible interstitial lung disease with unknown cause. To explore the role and regulatory mechanism of leucine-rich repeat-containing protein 15 (LRRC15) in IPF, bleomycin (BLM)-induced pulmonary fibrosis in mouse and A549 cells were constructed, and the expression of LRRC15 were detected. Then, MTT, GFP-RFP-LC3 dual fluorescent labeling system and Western blotting were used to investigate the effects of LRRC15 on cell activity and autophagy after transfection of siLRRC15, respectively. The results indicated that the expression of LRRC15 was significantly increased after the BLM treatment in mouse lung tissue and A549 cells. The designed and synthesized siLRRC15 followed by transfection into A549 cells resulted in a dramatic reduction in LRRC15 expression and partially restored the cell damage induced by BLM. Moreover, the expression of LC3-II and P62 were up-regulated, the amount of autophagosome were increased by GFP-RFP-LC3 dual fluorescent labeling assay after BLM treatment. Meanwhile, this study also showed that the key autophagy proteins LC3-II, ATG5 and ATG7 were up-regulated, P62 was down-regulated and autophagic flux were enhanced after further treatment of A549 cells with siLRRC15. The above findings suggest that LRRC15 is an indicator of epithelial cell damage and may participate in the regulation of fibrosis through autophagy mechanism in IPF. This study provides necessary theoretical basis for further elucidating the mechanism of IPF.

Structural Modification and Optimisation of Hyperoside Oriented to Inhibit TGF-β-Induced EMT Activity in Alveolar Epithelial Cells.

Pulmonary fibrosis (PF) is a disease characterised by diffuse nonspecific alveolar inflammation with interstitial fibrosis, which clinically manifests as dyspnoea and a significant decline in lung function. Many studies have shown that the epithelial-mesenchymal transition (EMT) plays a pivotal role in the pathogenesis of pulmonary fibrosis. Based on our previous findings, hypericin (Hyp) can effectively inhibit the process of the EMT to attenuate lung fibrosis. Therefore, a series of hyperoside derivatives were synthesised via modifying the structure of hyperoside, and subsequently evaluated for A549 cytotoxicity. Among these, the pre-screening of eight derivatives inhibits the EMT. In this study, we evaluated the efficacy of Z6, the most promising hyperoside derivative, in reversing TGF-β1-induced EMTs and inhibiting the EMT-associated migration of A549 cells. After the treatment of A549 cells with Z6 for 48 h, RT-qPCR and Western blot results showed that Z6 inhibited TGF-β1-induced EMTs in epithelial cells by supressing morphological changes in A549 cells, up-regulating E-cadherin (p < 0.01, p < 0.001), and down-regulating Vimentin (p < 0.01, p < 0.001). This treatment significantly reduced the mobility of transforming growth factor β1 (TGF-β1)-stimulated cells (p < 0.001) as assessed by wound closure, while increasing the adhesion rate of A549 cells (p < 0.001). In conclusion, our results suggest that hyperoside derivatives, especially compound Z6, are promising as potential lead compounds for treating pulmonary fibrosis, and therefore deserve further investigation.

Investigation of mallow oil-entrapped alginate/chitosan liposomes in human lung carcinoma cells

Nowadays, drug delivery systems are designed to increase therapeutic effectiveness and reduce cytotoxic effects. Thanks to the tannins, phenolic compounds, ascorbic acid and flavonoids found in their extracts, plants belonging to the Malvaceae family can be used as nutraceuticals in the treatment of many diseases. In this study, two different extractions of the Mallow plant were performed, alginate/chitosan encapsulations were synthesized and characterized. The potential antiproliferative activity of the extracts and suspended liposomes was examined in the A549 cell line. Characteristic peaks were detected in FT-IR analyzes proving that extract and liposome packaging took place. The entrapment efficiency and content of the extract in the capsules (liposome 1 and 2) are respectively; It was determined as 87.64%, 3.94%, 91.56% and 5,24%. It was determined that 90% of the entrapped extract was released after approximately 40 h in both liposome applications. IC50 values were calculated with the CCK-8 method. According to the results, IC50 values for extracts 1 and 2 were found to vary between 103 and 147 μg/mL in the first 24 h and the effects of the extract increased as a result of encapsulation. It was also determined that IC50 values decreased to concentrations of 41–39 μg/mL, respectively. Treatment of A549 cells with capsules resulted in dose- and time-dependent cytotoxicity. Over time, the capsule secreted more extract, and the extract released over time gradually increased in 48 h, causing more cell death.

Abstract 201: Morphological changes to the vulvar squamous carcinoma cell line, A431, resulting from clobetasol-induced upregulation of vimentin

Abstract Vimentin is recognized as an important marker in the epithelial-mesenchymal transition (EMT) of several cancer cell types. The overexpression of vimentin appears to coincide with increased tumor growth, migration, and poor prognosis. The vulvar cancer cell line, A431, is of squamous cell origin and displays hallmarks of a squamous cell including expression of cytokeratins 8 and 18 as well as E- and P-cadherin. We have found that treatment of A431 cells with the glucocorticoid, clobetasol, results in upregulation of vimentin which is accompanied by change to a more mesenchymal-like morphology. Despite this change in morphology, the cells continue to express cytokeratins 8 and 18. This suggests that the vimentin has a greater impact on cell morphology than does cytokeratins 8 and 18. However, we previously reported that the analogous glucocorticoid, dexamethasone, also downregulates E- and P-cadherin in these cells which may account for the observed change in morphology. Here we examine more closely the impact of vimentin on cellular morphology in the clobetasol treated A431 cells by both immunofluorescence microscopy and Raman imaging. Citation Format: Jani E. Lewis, Kia Haering, Kazushige Yokoyama. Morphological changes to the vulvar squamous carcinoma cell line, A431, resulting from clobetasol-induced upregulation of vimentin [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 201.

Abstract 2589: Novel workflow to generate natural killer cell sensitivity prediction model based on the epithelial mesenchymal transition related reprogramming at a single cell resolution

Abstract Single cell technology allows us to examine cell heterogeneity and identify rare cell populations; however, the high cost of single cell RNAseq limits its adoption into clinical and research environments. As an inexpensive alternative, immunofluorescent assays analyzing cell morphologies have been used in high content screening to examine cell phenotype and response to external stresses. Natural Killer cells (NK) are the bodies first line of defense and gained traction as a potential target in treating cancer. A study showed that cancer cells undergoing the process epithelial mesenchymal transition (EMT) increase their vulnerability to NK mediated cytotoxicity. We hypothesize that the EM morphology change can be correlated with NK sensitivity. Our goal is to develop an image-based, cost-effective, and efficient workflow capable of extracting cancer cell morphological features to predict its NK sensitivity at the single-cell level. To optimize this workflow, we use A549 cell line cells treated with transforming growth factor- β (TGF-β) as a representative models of cells with various EM phenotypes as NK sensitivities. We utilized a Cell Painting inspired novel image based live cell cytotoxicity assay. To capture cell morphological features, we optimized an organelle specific and live cell compatible staining panel that targets the nucleus, nucleic acid, cell membrane, F-actin, and mitochondria. The cell viability is assessed by Propidium Iodide, which stains dead cell nuclei. Our image analysis pipeline can extract the morphological features and track the cells of interest during the experiment and assess their viability. Subsequently, the morphology and viability data will be processed by a classification algorithm that could discern morphological features of cancer cells that indicate high NK vulnerability. We started with treating A549 cells with 5ng/mL TGF-β for 0, 24, 48, and 72 hours, and successfully applied the organelle staining panel. We observed distinct cell morphologies at each timepoint of TGF-β treatment, with cells shifting from a cobblestone (no TGF-β) to a spindle-like (72 hours TGF-β) morphology. Notably, the F-actin signal was significantly overexpressed in the 72 hours TGF-β treated A549 cells. A CellProfiler based preliminary image analysis pipeline was established for cell segmentation, cell features extraction, and cell viability tracking with quality control filters. Further optimization for cell segmentation, viability tracking, and classification algorithms will be needed to enhance the robustness of the workflow. Our established workflow demonstrates the feasibility of live cell functional assay with organelle staining. This workflow could be adopted to correlate and predict single tumor cell susceptibility to cytotoxic immune cells, with the goal of developing a therapeutic index. Citation Format: Yin Zou, Harrison Ball, Cade Harris, Nithya Ramnath, Venkateshwar Keshamouni, Sunitha Nagrath. Novel workflow to generate natural killer cell sensitivity prediction model based on the epithelial mesenchymal transition related reprogramming at a single cell resolution [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2589.

PIEZO1 acts as a cancer suppressor by regulating the ROS/Wnt/β-catenin axis.

PIEZO1 works differently in different cancers and at different stages of development. The objective of the current study was to explore the function and underlying mechanism of PIEZO1 in lung adenocarcinoma (LUAD) cells. Different LUAD cell lines were treated with PIEZO1 inhibitor (GsMTx4) and agonist (Yoda1), and the expression of PIEZO1 in LUAD cells was detected using real-time quantitative PCR (RT-qPCR) and western blotting. The effects of PIEZO1 on invasion, migration and epithelial-mesenchymal transition (EMT) markers protein expression of LUAD cells were detected using the MTT assay, flow cytometry, transwell assay, wound-healing assay, and western blotting. Reactive oxygen species (ROS) agonists (BAY 87-2243) and inhibitors (NAC) and Wnt/β-catenin pathway inhibitors (iCRT3) were selected to treat A549 cells to investigate the mechanism of PIEZO1 on ROS production and Wnt/β-catenin expression in A549 cells. In A549, NCI-H1395, and NCI-H1975 cells, GsMTx4 promoted cell proliferation, invasion, migration, upregulated EMT-related marker protein expression, and inhibited cell apoptosis, while Yoda1 exerted effects opposite to those of GsMTx4. In A549 cells, GsMTx4 can reduce ROS production, it also inhibited ROS production, apoptosis, and downregulated proapoptotic markers induced by BAY 87-2243. Importantly, BAY 87-2243 blocked the effect of GSMTX4-induced Wnt/β-catenin overexpression. Similarly, Yoda1 can reduce the effect of NAC. In addition, iCRT3 can block the upregulation of EMT-related marker proteins by GsMTx4, and increase apoptosis and decrease cell invasion and migration. In summary, PIEZO1 acts as a cancer suppressor by regulating the ROS/Wnt/β-catenin axis, providing a new perspective on the role of mechanosensitive channel proteins in cancer.

Mikania micrantha silver nanoparticles exhibit anticancer activities against human lung adenocarcinoma via caspase-mediated apoptotic cell death

Green-mediated synthesis of nanoparticles has earned a promising role in the area of nanotechnology due to their biomedical applications. This study describes the synthesis of silver nanoparticles (AgNPs) using Mikania micrantha leaf extract and its functional activities against cancer. The synthesis of AgNPs was confirmed using Ultraviolet-Visible (UV-Vis) spectrum that exhibited an absorption band at 459 nm. The bioactive compounds of M. micrantha leaf extract that functioned as reducing and capping agents were confirmed by a shift in the absorption bands in Fourier Transform Infra-red Spectroscopy (FT-IR). Scanning Electron Microscopy (SEM) and Transmission Electron Microscopy (TEM) studies validated the spherical shape and size of AgNPs, respectively. Energy Dispersive Spectroscopy (EDS) analysis revealed the presence of elemental silver. The crystalline nature of AgNPs was confirmed by the X-ray Diffraction Analysis (XRD). AgNPs effectively induced cytotoxicity and prevented A549 cell colony formation in a dose-dependent manner. Treatment of A549 cells with AgNPs also increased DNA damage, which was coupled with elevated lipid peroxidation and decreased antioxidant enzymes such as glutathione (GSH), glutathione-s-transferase (GST), and superoxide dismutase (SOD). Following AgNPs treatment, the mRNA expression levels of the pro-apoptotic genes as well as the activities of caspases were significantly elevated in A549 cells while the expression levels of anti-apoptotic genes were downregulated. Our study demonstrates the potential of the synthesised AgNPs for cancer therapy possibly targeting the apoptotic pathway.

Fufang Luohanguo Qingfei granules reduces influenza virus susceptibility via MAVS-dependent type I interferon antiviral signaling

Ethnopharmacological relevanceFufang Luohanguo Qingfei granules (LQG) is a Chinese patent medicine, clinically used to treat flu-like symptoms including cough with yellow phlegm, impeded phlegm, dry throat and tongue. However, the protective activity of LQG against influenza infection is indeterminate. Aim of the studyThis study is to investigate the therapeutic effect of LQG on influenza infection and elucidate its underlying mechanism. Materials and methodsIn vivo: A viral susceptible mouse model induced by restraint stress was established to investigate LQG's beneficial effects on influenza susceptibility. MAVS knockout (Mavs−/−) mice were used to verify the potential mechanism of LQG. In vitro: Corticosteroid (CORT)-treated A549 cells were employed to identify the active ingredients in LQG. Mice morbidity and mortality were monitored daily for 21 days. Histopathologic changes and inflammatory cytokines in lung tissues were examined by H&E staining and ELISA. RNA-seq was used to explore the signaling pathway influenced by LQG and further confirmed by qPCR. Immunoblotting and immunohistochemistry (IHC) were used to determine the protein levels. CO-IP and DARTS were applied to detect protein-protein interaction and compound-protein interaction, respectively. ResultsLQG effectively attenuated the susceptibility of restrained mice to H1N1 infection. LQG significantly boosted the production of IFN-β transduced by mitochondrial antiviral-signaling protein (MAVS), while MAVS deficiency abrogated its protective effects on restrained mice infected with H1N1. Moreover, in vitro studies further revealed that mogroside Ⅱ B, amygdalin, and luteolin are potentially active components of LQG. ConclusionThese results suggested that LQG inhibited the mitofusin 2 (Mfn2)-mediated ubiquitination of MAVS by impeding the E3 ligase synoviolin 1 (SYVN1) recruitment, thereby enhancing IFN-β antiviral response. Overall, our work elaborates a potential regimen for influenza treatment through reduction of stress-induced susceptibility.

Probenecid Inhibits Influenza A(H5N1) and A(H7N9) Viruses In Vitro and in Mice.

Avian influenza (AI) viruses cause infection in birds and humans. Several H5N1 and H7N9 variants are highly pathogenic avian influenza (HPAI) viruses. H5N1 is a highly infectious bird virus infecting primarily poultry, but unlike other AIs, H5N1 also infects mammals and transmits to humans with a case fatality rate above 40%. Similarly, H7N9 can infect humans, with a case fatality rate of over 40%. Since 1996, there have been several HPAI outbreaks affecting humans, emphasizing the need for safe and effective antivirals. We show that probenecid potently inhibits H5N1 and H7N9 replication in prophylactically or therapeutically treated A549 cells and normal human broncho-epithelial (NHBE) cells, and H5N1 replication in VeroE6 cells and mice.

Combination of the cuproptosis inducer disulfiram and anti‑PD‑L1 abolishes NSCLC resistance byATP7B to regulate the HIF‑1 signaling pathway.

Disulfiram (DSF) is used to treat non‑small cell lung cancer (NSCLC). DSF significantly increases expression of programmed death‑ligand 1 (PD‑L1), which may enhance immunosuppression and immune escape of tumors. Therefore, the present study aimed to investigate the role of combined treatment of DSF and anti‑PD‑L1 in NSCLC resistance. The viability and apoptosis of A549 cells were detected by the Cell Counting Kit‑8 assay and flow cytometry, respectively. The expression levels of ATPase copper‑transportingβ (ATP7B) and PD‑L1 in A549 cells were detected by reverse transcription‑quantitative PCR and western blot analysis. The levels of reactive oxygen species (ROS), malondialdehyde (MDA) and superoxide dismutase (SOD) in A549 cells were detected by respective assay kits. The expression levels of cuproptosis‑associated proteins ferredoxin‑1 (FDX1), ATP7B, solute carrier family 31 member 1 (SLC31A1), succinate dehydrogenase B (SDHB), PD‑L1 and hypoxia inducible factor (HIF)‑1A were analyzed by western blotting in A549 cells. DSF inhibited the viability of A549 cells and promoted expression levels of ATP7B and PD‑L1 at both mRNA and protein levels in A549 cells. The viability of cisplatin (DPP)‑treated A549 cells was increased following DSF treatment. JQ‑1 (a PD‑L1 inhibitor) suppressed the viability of DPP‑treated A549 cells pretreated with DSF. DSF increased expression levels of ATP7B and PD‑L1. The combination treatment of DSF and JQ‑1 in A549 cells increased levels of ROS and MDA, as well as expression levels of FDX1 and SLC31A1; however, combination treatment decreased levels of SOD, as well as expression levels of ATP7B, SDHB, PD‑L1, and HIF‑1A. PX478 (an HIF‑1 inhibitor) acted with DSF to enhance the inhibitory effects on the viability and on the induction of apoptosis of A549 cells. PX478 upregulated the levels of ROS and MDA, while it downregulated levels of SOD in DSF‑treated A549 cells. PX478 promoted expression levels of FDX1 and SLC31A1, while it suppressed expression levels of ATP7B, PD‑L1, and HIF‑1A in DSF‑treated A549 cells. In conclusion, the combined treatment of A549 cells with anti‑PD‑L1 and DSF enhanced the effect of cuproptosis on the inhibition of NSCLC cell viability.

Luteolin Alleviates Oxidative Stress in Chronic Obstructive Pulmonary Disease Induced by Cigarette Smoke via Modulation of the TRPV1 and CYP2A13/NRF2 Signaling Pathways.

The current study aims to investigate the therapeutic potential of luteolin (Lut), a naturally occurring flavonoid found in various medicinal plants, for treating chronic obstructive pulmonary disease (COPD) through both in vitro and in vivo studies. The results demonstrated that Lut increased body weight, reduced lung tissue swelling and lung damage indices, mitigated systemic oxidative stress levels, and decreased alveolar fusion in cigarette smoke (CS)- and lipopolysaccharide (LPS)-induced COPD mice. Additionally, Lut was observed to downregulate the expression of the TRPV1 and CYP2A13 proteins while upregulating SIRT6 and NRF2 protein expression in CS + LPS-induced COPD mice and cigarette smoke extract (CSE)-treated A549 cells. The concentrations of total reactive oxygen species (ROS) and mitochondrial ROS in A549 cells induced by CSE significantly increased. Moreover, CSE caused a notable elevation of intracellular Ca2+ levels in A549 cells. Importantly, Lut exhibited inhibitory effects on the inward flow of Ca2+ and attenuated the overproduction of mitochondrial and intracellular ROS in A549 cells treated with CSE. In conclusion, Lut demonstrated a protective role in alleviating oxidative stress and inflammation in CS + LPS-induced COPD mice and CSE-treated A549 cells by regulating TRPV1/SIRT6 and CYP2A13/NRF2 signaling pathways.

Role of the cAMP-PKA-NF-κB pathway in Mucin1 over-expression in A549 cells during Respiratory syncytial virus infection

Respiratory syncytial virus (RSV) is the most common pathogen associated with acute lower respiratory tract infections in infants and young children worldwide. RSV commonly presents as bronchiolitis in young children; however, it can sometimes progress to pneumonia, respiratory failure, apnoea and even death. Although mucin1 (MUC1), a type of transmembrane glycoprotein present on airway epithelial surfaces, plays a crucial anti-inflammatory role in airway infections; however, its roles in RSV-associated acute lower respiratory tract infections have rarely been explored. In this study, we first revealed very high MUC1 protein levels in the exacerbation phase in sputum samples from children with RSV bronchiolitis. Because MUC1 is the downstream target of tumour necrosis factor-alpha (TNF-α) in RSV-infected A549 cells, we observed the inhibition of NF-κB activity, main downstream signalling of TNF-α and remarkably reduced levels of MUC1 in RSV-infected and TNF-α treated A549 cells. Furthermore, the cyclic adenosine monophosphate (cAMP) analogue (dbcAMP) downregulated the protein levels of p-IκBα and MUC1 in TNF-α-treated A549 cells. By contrast, a protein kinase A inhibitor (KT5720) up-regulated the levels of those proteins. dbcAMP and KT5720 had the same effects on MUC1 protein levels in RSV-infected A549 cells. In conclusion, we found that the cAMP-PKA-NF-κB pathway may play a role in the regulation of MUC-1 over-expression during RSV infection.

Near-infrared absorption and emission probes with optimal connection bridges for live monitoring of NAD(P)H dynamics in living systems

Two NAD(P)H-biosensing probes consisting of 1,3,3-trimethyl-3H-indolium and 3-quinolinium acceptors, linked by thiophene, A, and 3,4-ethylenedioxythiophene, B, bridges are detailed. We synthesized probes C and D, replacing the thiophene connection in probe A with phenyl and 2,1,3-benzothiadiazole units, respectively. Probe E was prepared by substituting probe A's 3-quinolinium unit with a 1-methylquinoxalin-1-ium unit. Probe solutions are non-fluorescent but in the presence of NADH, exhibit near-infrared fluorescence at 742.1 nm and 727.2 nm for probes A and B, respectively, and generate absorbance signals at 690.6 nm and 685.9 nm. In contrast, probes C and D displayed pronounced interference from NADH fluorescence at 450 nm, whereas probe E exhibited minimal fluorescence alterations in response to NAD(P)H. Pre-treatment of A549 cells with glucose in the presence of probe A led to a significant increase in fluorescence intensity. Additionally, subjecting probe A to lactate and pyruvate molecules resulted in opposite changes in NAD(P)H levels, with lactate causing a substantial increase in fluorescence intensity, conversely, pyruvate resulted in a sharp decrease. Treatment of A549 cells with varying concentrations of the drugs cisplatin, gemcitabine, and camptothecin (5, 10, and 20 µM) led to a concentration-dependent increase in intracellular fluorescence intensity, signifying a rise in NAD(P)H levels. Finally, fruit fly larvae were treated with different concentrations of NADH and cisplatin illustrating applicability to live organisms. The results demonstrated a direct correlation between fluorescence intensity and the concentration of NADH and cisplatin, respectively, further confirming the efficacy of probe A in sensing changes in NAD(P)H levels within a whole organism.

Dinuclear gold(I) complexes based on carbene and diphosphane ligands: bis[2-(dicyclohexylphosphano)ethyl]amine complex inhibits the proteasome activity, decreases stem cell markers and spheroid viability in lung cancer cells.

Three new dinuclear gold(I) complexes (1-3) containing a carbene (1,3-Bis(2,6-di-isopropylphenyl)imidazol-2-ylidene (IPr)) and diphosphane ligands [bis(1,2-diphenylphosphano)ethane (Dppe), bis(1,3-diphenylphosphano)propane (Dppp) and bis[2-(dicyclohexylphosphano)ethyl]amine (DCyPA)], were synthesized and characterized by elemental analysis and, ESI-MS, mid FT-IR and NMR spectroscopic methods. The structures of complexes 2 and 3 were determined by X-ray crystallography, which revealed that the complexes are dinuclear having gold(I) ions linearly coordinated. The anticancer activities of the complexes (1-3) were evaluated in lung (A549), breast (MC-F7), prostate (PC-3), osteosarcoma (MG-63) and ovarian (A2780 and A2780cis) cancer models. Growth inhibition by the new complexes was higher than cisplatin in all cell lines tested. The mechanism of action of complex 3 was investigated in A549 cells using 2-dimensional (2D) models and 3D-multicellular tumor spheroids. Treatment of A549 cells with complex 3 caused: the induction of apoptosis and the generation of reactive oxygen species; the cell cycle arrest in the G0/G1 phase; the inhibition of both the proteasome and the NF-kB activity; the down-regulation of lung cancer stem cell markers (NOTCH1, CD133, ALDH1 and CD44). Complex 3 was more active than cisplatin also in 3D models of A549 lung cancer cells.

SUSTAINED ANTICANCER EFFECT BY NARINGIN-LOADED ZINC OXIDE NANOPARTICLES IN HUMAN LUNG ADENOCARCINOMA A549 CELLS

Objective: This study aimed to design naringin-loaded zinc oxide nanoparticles (Nar-ZnO NPs) and evaluate the formulated nanoparticles (NPs) for their antioxidant and anticancer potential. Methods: Naringin-loaded zinc oxide nanoparticles (Nar-ZnO NPs) were prepared using a modified sol-gel method with ethylenediaminetetraacetic acid (EDTA) as a capping agent. Subsequently, were characterized using dynamic light scattering (DLS), powder X-ray diffraction (PW-XRD), Fourier transform infrared spectroscopy (FT-IR), high-resolution scanning electron microscopy (HR-SEM), and Energy Dispersive X-ray analysis (EDX). Furthermore, the naringin-loaded zinc oxide nanoparticles (Nar-ZnO NPs) were evaluated for their in vitro free radical scavenging activity using antioxidant assays and inhibition of lipid peroxidation potential using the altered thiobarbituric acid-reactive species (TBARS) test. The cytotoxic effect of naringin-loaded zinc oxide nanoparticles (Nar-ZnO NPs) on the non-transformed Vero cell line and lung cancer A549 cell line was investigated using the (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) MTT assay. Apoptosis study was conducted using the Acridine orange/Ethidium bromide (AO/EB) double staining assay, while propidium iodide (PI) stain was utilized to observe apoptotic morphological changes. Results: The prepared naringin-loaded zinc oxide nanoparticles (Nar-ZnO NPs) were smooth and hexagonal, with an average particle size of 500 nm. The antioxidant assays demonstrated that the naringin-loaded zinc oxide nanoparticles (Nar-ZnO NPs) and ascorbic acid exhibited comparable free radical scavenging and inhibition of lipid peroxidation activity. In MTT assay, the naringin-loaded zinc oxide nanoparticles (Nar-ZnO NPs) displayed IC₅₀ values of 1014.05 µg/ml for Vero cell lines and 317.51 µg/ml for A549 cells, highlighting their influence on cell viability. Remarkably, treatment of A549 cells with the Nar-ZnO NPs resulted in dose-dependent apoptotic morphological changes, as observed through (AO/EB) double staining assay and propidium iodide (PI) stain. Conclusion: The study findings revealed that the naringin-loaded zinc oxide nanoparticles (Nar-ZnO NPs) displayed dose-dependent free radical scavenging activity, significant inhibition of lipid peroxidation, and notable anticancer properties against A549 cells.