NCI-H292 Cells Research Articles

Cigarette smoke and extracellular Hsp70 induce secretion of ATP and differential activation of NLRP3 inflammasome in monocytic and bronchial epithelial cells

BackgroundChronic obstructive pulmonary disease (COPD) is an inflammatory disease mainly caused by smoking. Cigarette smoke damages airway epithelium and activates lung macrophages, causing inflammatory responses. It was suggested that nucleotide-binding oligomerization domain-like receptor family pyrin domain-containing 3 (NLRP3) inflammasome might have an important role in COPD development.Study aimed to explore whether cigarette smoke extract (CSE), extracellular heat shock protein 70 (eHsp70) or their combinations induce adenosine triphosphate (ATP) release and NLRP3 inflammasome activation. MethodsWe detected NLRP3 and interleukin (IL)-1β mRNA expression, extracellular IL-1β and ATP concentrations as well as lactate dehydrogenase (LDH) activity. We used bronchial epithelial (NCI-H292, 16HBE and NHBE) and monocytic cells (monocyte-derived macrophages (MDMs) and THP-1) as representative of local airway and systemic compartments that could be affected in COPD. ResultsCSE and eHsp70 increased NLRP3 and IL-1β mRNA expression as well as IL-1β and ATP secretion in all cells compared to untreated cells. Lytic cell death was observed in cell lines, especially those of bronchial epithelium origin, but not in primary cells (NHBE, MDMs). Regarding LDH activity, eHsp70 did not modulate CSE effects, except in NCI-H292 cell line. However, eHsp70 significantly affected CSE-provoked NLRP3 inflammasome activation by causing mostly antagonistic effects in airway epithelial cells and synergistic effects in MDMs. ConclusionWe demonstrated that both CSE and eHsp70 induce ATP secretion and differential activation of NLRP3 inflammasome in bronchial epithelial and monocytic cells. We suggest that these mechanisms might be involved in pathophysiology of COPD by contributing to the propagation of inflammation.

English

Cassia grandis L. (Fabaceae), a native tree from Amazon Forest, has been used in folk medicine against worms and intestinal parasites, and to treat stomach and respiratory problems, blood diseases, among others uses. This study aimed to evaluate the antioxidant, cytotoxic, antimicrobial, and schistosomicidal activities of the methanolic extract from C. grandis stem bark (CgME) and to elucidate the chemical profile of its active fractions. The antioxidant activity of C. grandis stem bark methanolic extract (CgME) and its fractions were determined by DPPH radical scavenging assay and by the total phenolics and flavonoid contents. The antimicrobial activity was performed by microdilution. The cytotoxicity against MCF-7 (breast), NCI-H292 (lung), and HL-60 (leukemia) cancer cell lines was evaluated by the MTT method. The schistosomicidal activity was investigated in vitro against adult Schistosoma mansoni couple worms. The phytochemical profile of the active fractions was determined by GC-MS and UPLC-MS. The hexane fraction from the CgME was cytotoxic to NCI-H292 and HL-60 cancer cell lines and both major compounds clionasterol and lupeol acetate, determined by GC-MS, are well known for their cytotoxicity against cancer cells. The ethyl acetate fraction (CgEF) exhibited both antibacterial, against multidrug-resistant S. aureus, and schistosomicidal activities, which could be attributed to the presence of flavonoids, such as catechin derivatives, quercetin, and luteolin in the CgEF. These results agree with the popular uses of C. grandis and should stimulate future research on this species. Key words: Antimicrobial activity, antioxidant, Staphylococcus aureus, clinical isolates, Schistosoma mansoni, cytotoxicity. &nbsp

Oxysophocarpine protects airway epithelial cells against inflammation and apoptosis by inhibiting miR-155 expression.

Oxysophocarpine (OSC) has been documented for anti-inflammatory activity. However, the mechanisms of OSC in anti-inflammation are unclear. Aim: To investigate the protective effects of OSC on inflammation and apoptosis induced by lipopolysaccharide in NCI-H292 and human primary airway epithelial cells. Materials & methods: MTT and Annexin V-FITC/PI staining were used to detect cells viability. Inflammatory responses were determined by ELISA. The quantitative real-time PCR (qRT-PCR) and western blot were used to detect mRNA/miRNA and protein expressions respectively. Co-immunoprecipitation was investigated for protein interactions. Results & conclusion: miR-155 mimics significantly induced cell apoptosis, inflammatory responses and MAPK and NF-κB pathways. NDFIP1 was identified as the target of miR-155. OSC protected cells against apoptosis and inflammatory responses and compromised miR-155 activity by attenuating MAPK and NF-κB pathways.

Lindera obtusiloba Attenuates Oxidative Stress and Airway Inflammation in a Murine Model of Ovalbumin-Challenged Asthma.

Lindera obtusiloba is widespread in northeast Asia and used for treatment of improvement of blood circulation and anti-inflammation. In this study, we investigated anti-inflammatory and anti-oxidant effects of the methanolic extract of L. obtusiloba leaves (LOL) in an ovalbumin (OVA)-challenged allergic asthma model and tumor necrosis factor (TNF)-α-stimulated NCI-H292 cell. Female BALB/c mice were sensitized with OVA by intraperitoneal injection on days 0 and 14, and airway-challenged with OVA from days 21 to 23. Mice were administered 50 and 100 mg/kg of LOL by oral gavage 1 h before the challenge. LOL treatment effectively decreased airway hyper-responsiveness and inhibited inflammatory cell recruitment, Th2 cytokines, mucin 5AC (MUC5AC) in bronchoalveolar lavage fluid in OVA-challenged mice, which were accompanied by marked suppression of airway inflammation and mucus production in the lung tissue. LOL pretreatment inhibited the phosphorylation of mitogen-activated protein kinases (MAPKs) and nuclear factor-kappa B (NF-κB) with suppression of activator protein (AP)-1 and MUC5AC in the lung tissue. LOL also down-regulated expression of inflammatory cytokines, and inhibited the activation of NF-κB in TNF-α-stimulated NCI-H292 cells. LOL elevated the translocation of nuclear factor-erythroid 2-related factor (Nrf-2) into nucleus concurrent with increase of heme oxyngenase-1 (HO-1) and NAD(P)H quinine oxidoreductase 1 (NQO1). Moreover, LOL treatment exhibited a marked increase in the anti-oxidant enzymes activities, whereas effectively suppressed the production of reactive oxygen species and nitric oxide, as well as lipid peroxidation in lung tissue of OVA-challenged mice and TNF-α-stimulated NCI-H292 cells. These findings suggest that LOL might serve as a therapeutic agent for the treatment of allergic asthma.

RETRACTED ARTICLE: Shp2 positively regulates cigarette smoke-induced epithelial mesenchymal transition by mediating MMP-9 production

Cigarette smoke (CS) is a major risk factor for the development of lung cancer and chronic obstructive pulmonary disease (COPD). Epithelial-mesenchymal transition (EMT) commonly coexists in lung cancer and COPD. CS triggers many factors including matrix metalloproteinases (MMPs) production, contributing to EMT progression in the lungs. Here, how Shp2 signaling regulates the CS-induced MMP-9 production and EMT progression were investigated in mouse lungs and in pulmonary epithelial cell cultures (NCI-H292) found CS induced MMP-9 production, EMT progression (increased vimentin and α-SMA; decreased E-cadherin) and collagen deposition in lung tissues; cigarette smoke extract (CSE) induced MMP-9 production and EMT-related phenotypes in NCI-H292 cells, which were partially prevented by Shp2 KO/KD or Shp2 inhibition. The CSE exposure induced EMT phenotypes were suppressed by MMP-9 inhibition. Recombinant MMP-9 induced EMT, which was prevented by MMP-9 inhibition or Shp2 KD/inhibition. Mechanistically, CS and CSE exposure resulted in ERK1/2, JNK and Smad2/3 phosphorylation, which were suppressed by Shp2 KO/KD/inhibition. Consequentially, the CSE exposure-induced MMP-9 production and EMT progression were suppressed by ERK1/2, JNK and Smad2/3 inhibitors. Thus, CS induced MMP-9 production and EMT resulted from activation of Shp2/ERK1/2/JNK/Smad2/3 signaling pathways. Our study contributes to the underlying mechanisms of pulmonary epithelial structural changes in response to CS, which may provide novel therapeutic solutions for treating associated diseases, such as COPD and lung cancer.

Synthesis and characterization of novel naphthalene-derivatized tridentate ligands and their net neutral rhenium tricarbonyl complexes and cytotoxic effects on non-small cell lung cancer cells of interest

Rhenium complexes have possible fluorescence imaging, radioimaging and therapeutic applications and can also serve as a model system for 99mTc. The quest for new pharmaceutical agents is thus an expanding area of research. In this study, we report two net neutral complexes, [Re(CO)3(N(SO2)(1-nap)dien)] and [Re(CO)3(N(SO2)(2-nap)dien)], synthesized by treating fac-[Re(CO)3(H2O)3]+ with two novel tridentate ligands, (N(SO2)(1-nap)dienH and N(SO2)(2-nap)dienH), derived from diethylenetriamine (dien) attached to a sulfonamide group. The compounds were characterized by UV–Visible, FTIR, 1H NMR and fluorescence spectroscopies, together with X-ray crystallography. It is evident from the 1H NMR spectroscopic data recorded in DMSO‑d6 as well as X-ray crystallography supported by FTIR data, that upon coordination to the metal centre, the sulfonamide nitrogen atom is deprotonated and the ligand binds with the metal in a tridentate fashion. Both the complexes have a distorted octahedral structure where the Re(I) metal is coordinated by three nitrogen atoms of the dien backbone and three CO ligands. Emission spectra were recorded in methanol and an enhanced fluorescence emission was observed in the 330–345 nm range for the ligands while the corresponding Re complexes showed no or quenched fluorescence intensities. The in vitro cytotoxic activity of the synthesized compounds was examined using NCI-H292 (non-small cell lung cancer cells) and MRC-5 (the human normal lung fibroblast cell line). The highest cytotoxic activity was observed for the [Re(CO)3(N(SO2)(1-nap)dien)] complex on NCI-H292 cells with an IC50 value of 9.91 μM at 48 h. However all four compounds show acute cytotoxicity for MRC-5 cells at 24 h. The promising cytotoxic activity of the novel ligands and metal complexes indicate that these compounds may be good candidates to be utilized as anticancer drug leads against lung cancer.

Three Tiaobu Feishen formulae reduces cigarette smoke-induced inflammation in human airway epithelial cells.

To investigate the therapeutic efficacy of Tiaobu Feishen formulae (TBFS) on cigarette smoke-induced inflammation in vitro using lipopolysaccharide (LPS)-induced and cigarette smoke extract (CSE)-induced NCI-H292 cells. We evaluated the inhibitory effects of Bufei Jianpi formula (BJF), Bufei Yishen formula (BYF), and Yiqi Zishen formula (YZF) on the expressions of inflammatory cytokines including tumor necrosis factor (TNF)-α and interleukin (IL)-8, matrix metalloproteinase (MMP)-9, tissue inhibitor of matrix metalloprotease (TIMP)-1, and superoxide dismutase (SOD) in H292 cells stimulated with LPS or CSE. Their related transcription factors and signaling pathways were also analyzed. BJF, BYF, and YZF significantly inhibited the LPS- or CSE-induced expressions of TNF-α, IL-8, MMP-9, TIMP-1, and SOD in H292 cells, and suppressed the activation of transcription factors including nuclear transcription factor (NF)-κB, activator protein (AP)-1, and signal transducers and activators of transcription (STAT) 3 and their corresponding pathways, including NF-κB, mitogen-activated protein kinase (MAPK), STAT3, and peroxisome proliferator-activated receptor (PPAR). BJF, BYF, and YZF effectively suppressed inflammatory responses, protease-antiprotease imbalance, and oxidative stress induced by LPS and CSE, an effect that was closely associated with the inhibition of the NF-κB, MAPK, STAT3, and PPAR pathways.

CDC20 and TRIB3 Upregulated by TFAP2C Facilitate Cell Proliferation in Non‐small Cell Lung Cancer Cells

In recent years, non‐small cell lung cancer (NSCLC) is the most serious cause of global cancer‐related mortality. To resolve this circumstance, many studies for identification of novel molecular targets have been conducted. In our previous investigation, a microarray analysis that associated with transcription factor activating enhancer‐binding protein 2C (TFAP2C) was carried out and we found out TFAP2C's oncogenic roles in NSCLC development. In the current study, we intended to reveal novel biomarkers for NSCLC, especially focused on several oncogenes transcriptionally regulated by TFAP2C. With the result of microarray analysis, cell division cycle 20 (CDC20) and tribbles pseudokinase 3 (TRIB3) were selected as promising oncogenes by transcriptionally upregulated by TFAP2C. In the treatment of TFAP2C‐specific siRNA in two NSCLC cell lines (NCI‐H292 and NCI‐H838 cells), we observed that mRNA and protein levels of CDC20 and TRIB3 were down‐regulated. In addition, when CDC20 or TRIB3 was overexpressed, cell viability was increased and reversely apoptosis of these cells was suppressed. Consequently, these results indicated that NSCLC tumorigenesis affected by two promising oncogenes, CDC20 and TRIB3, and they could act as potent markers to prognosis of NSCLC.Support or Funding InformationThis work was supported by the National Research Foundation of Korea (NRF) funded by the Ministry of Science and ICT (2019R1C1C1009423).

An efficient and high-yielding method for extraction and purification of linamarin from Cassava; in vitro biological evaluation

During the last three decades, studies of linamarin extracted from cassava have received increased attention due to the presence of high cyanogenic compounds in these extracts. The methods that are utilized to isolate linamarin are either tedious or use acidic conditions resulting in poor yields. In this study, a novel cryocooled method of extraction has been developed to isolate linamarin from Cassava root peel. Approximately 18 g of linamarin was isolated from 1 kg of fresh Cassava root peel, which is the highest amount reported to date. Linamarin was fully characterized using NMR, IR and LCMS. The anti-cancer properties of pure linamarin and Cassava crude extract were evaluated by a comprehensive cytotoxic assay, using MCF-7, HepG2, NCI H-292, AN3CA and MRC-5 cell lines. The crude extract showed higher cytotoxicity compared to pure linamarin. The results of the biological evaluation are comparable to other reported studies in the literature.

Cigarette smoke and electronic cigarettes differentially activate bronchial epithelial cells

BackgroundThe use of electronic cigarettes (ECIGs) is increasing, but the impact of ECIG-vapor on cellular processes like inflammation or host defense are less understood. The aim of the present study was to compare the acute effects of traditional cigarettes (TCIGs) and ECIG-exposure on host defense, inflammation, and cellular activation of cell lines and primary differentiated human airway epithelial cells (pHBE).MethodsWe exposed pHBEs and several cell lines to TCIG-smoke or ECIG-vapor. Epithelial host defense and barrier integrity were determined. The transcriptome of airway epithelial cells was compared by gene expression array analysis. Gene interaction networks were constructed and differential gene expression over all groups analyzed. The expression of several candidate genes was validated by qRT-PCR.ResultsBacterial killing, barrier integrity and the expression of antimicrobial peptides were not affected by ECIG-vapor compared to control samples. In contrast, TCIGs negatively affected host defense and reduced barrier integrity in a significant way. Furthermore ECIG-exposure significantly induced IL-8 secretion from Calu-3 cells but had no effect on NCI-H292 or primary cells. The gene expression based on array analysis distinguished TCIG-exposed cells from ECIG and room air-exposed samples.ConclusionThe transcriptome patterns of host defense and inflammatory genes are significantly distinct between ECIG-exposed and TCIG-treated cells. The overall effects of ECIGs on epithelial cells are less in comparison to TCIG, and ECIG-vapor does not affect host defense. Nevertheless, although acute exposure to ECIG-vapor induces inflammation, and the expression of S100 proteins, long term in vivo data is needed to evaluate the chronic effects of ECIG use.

Cu(II)-Coordination Polymer Nanostructures Induce Human Lung Cancer Cell Carcinoma Apoptotic Cell Death In Vitro and In Vivo

In the research, two novel coordination polymers based on Cu(II) ion as the metal source and their nanostructures have been prepared via utilizing two ligands of positional isomeric pyridine carboxylic acid 2-(5-carboxypyridin-3-yl)terephthalic acid (H3L2) and 4-(6-carboxypyridin-3-yl)phthalic acid (H3L1) under the conditions of ultrasonic and solvothermal reaction, and their chemical formulas are {[Cu3(L1)2(H2O)6](H2O)7}n (1) along with {[Cu3(L1)2(H2O)2](H2O)2}n (2). In addition, the treatment effect of 1 and 2 on the human cancer in vitro and in vivo was detected. The Cell Counting Kit-8 (CCK8) method was used to determine the proliferation of human lung cancer cell line (NCI-H292) after nanoparticles 1–2 treatment, the results exhibited that nanoparticle 1 had excellent inhibitory function on NCI-H292 cell viability. Besides, ROS accumulation in the NCI-H292 cells was detected by reactive oxygen species (ROS) assay and annexin V-FITC/PI assay was conducted to detect the apoptotic cells numbers of NCI-H292 cells induced by nanoparticles 1–2 treatment. In addition, the tumor model was established in vivo and the treatment ability of the nanoparticle on NCI-H292 tumor was evaluated by measuring the mice weight and tumor volume.

Spiraea prunifolia var. simpliciflora Attenuates Oxidative Stress and Inflammatory Responses in a Murine Model of Lipopolysaccharide-Induced Acute Lung Injury and TNF-α-Stimulated NCI-H292 Cells.

Spiraea prunifolia var. simpliciflora (SP) is traditionally used as an herbal remedy to treat fever, malaria, and emesis. This study aimed to evaluate the anti-oxidative and anti-inflammatory properties of the methanol extract of SP leaves in tumor necrosis factor (TNF)-α-stimulated NCI-H292 cells and in a lipopolysaccharide (LPS)-induced acute lung injury (ALI) mouse model. SP decreased the number of inflammatory cells and the levels of TNF-α, interleukin (IL)-1β, and IL-6 in the bronchoalveolar lavage fluid, and inflammatory cell infiltration in the lung tissues of SP-treated mice. In addition, SP significantly suppressed the mRNA and protein levels of TNF-α, IL-1β, and IL-6 in TNF-α-stimulated NCI-H292 cells. SP significantly suppressed the phosphorylation of the mitogen-activated protein kinases (MAPKs) and p65-nuclear factor-kappa B (NF-κB) in LPS-induced ALI mice and TNF-α-stimulated NCI-H292 cells. SP treatment enhanced the nuclear translocation of nuclear factor erythroid 2-related factor (Nrf2) with upregulated antioxidant enzymes and suppressed reactive oxygen species (ROS)-mediated oxidative stress in the lung tissues of LPS-induced ALI model and TNF-α-stimulated NCI-H292 cells. Collectively, SP effectively inhibited airway inflammation and ROS-mediated oxidative stress, which was closely related to its ability to induce activation of Nrf2 and inhibit the phosphorylation of MAPKs and NF-κB. These findings suggest that SP has therapeutic potential for the treatment of ALI.

Evaluation of anticancer effects of a pharmaceutically viable extract of a traditional polyherbal mixture against non-small-cell lung cancer cells

ObjectiveThe present work tested organic solvents to prepare an extract with anticancer properties from a polyherbal mixture containing Nigella sativa (seeds), Hemidesmus indicus (roots) and Smilax glabra (rhizomes). We evaluate anticancer effects in non-small-cell lung cancer cells (NCI-H292), and discuss optimization for pharmaceutical use in the context of efficacy, yield and toxicity. MethodsUsing different organic solvents, six extracts were prepared from the polyherbal mixture. Based on the cytotoxic effects of these extracts on NCI-H292 cells and normal lung cells (MRC-5), as evaluated by the sulphorhodamine B assay, the total ethyl acetate (T-EA) extract was selected for further analysis. The possible anticancer mechanisms were assessed by evaluating the extract’s effects on apoptosis (through fluorescent microscopic analysis, DNA fragmentation analysis, caspase 3/7 assay and analysis of expression levels of apoptosis-related genes p53, Bax, survivin, Hsp70 and Hsp90), colony formation and antioxidant activity. ResultsThe extract had cytotoxic effects against NCI-H292 cells in a time- and dose-dependent manner. Significant antioxidant activity and inhibition of colony formation were also observed. The expression level of caspase 3/7 significantly (P < 0.001) increased in NCI-H292 cells treated with 50 μg/mL of the extract. The same dosage led to a significant increase in expression levels of Bax and p53 (P < 0.05 and P < 0.01 respectively), accompanied by a significant decrease (P < 0.0001) in survivin, Hsp70 and Hsp90. ConclusionT-EA extract of the above polyherbal mixture has cytotoxicity against NCI-H292 cells via induction of apoptosis, antioxidant effects and inhibition of colony formation.

Silica dioxide nanoparticles aggravate airway inflammation in an asthmatic mouse model via NLRP3 inflammasome activation

Silica dioxide nanoparticles (SiONPs) are mainly used in the rubber industry; however, they are a major air pollutant in Asia. Thus, extensive research on this issue is required. In this study, we investigated the effects of SiONPs on asthma aggravation and elucidated the underlying mechanism using ovalbumin (OVA)-induced asthmatic mice model and in NCI–H292 cells. Mice exposed to SiONPs showed markedly increased Penh values, inflammatory cell counts, and inflammatory cytokine levels compared to OVA-induced asthmatic mice. Exposure to SiONPs also induced additional airway inflammation and mucus secretion with increases in protein expression levels of thioredoxin-interacting protein (TXNIP), NOD-like receptor pyrin domain-containing 3 (NLRP3) inflammasome, and interleukin (IL)-1β compared to those in OVA-induced asthmatic mice. Treatment of SiONPs in NCI–H292 cells also significantly increased mRNA expression levels of inflammatory cytokines accompanied with elevation in the levels of TXNIP, NLRP3 inflammasome, and IL-1β proteins in a concentration-dependent manner. Taken together, exposure to SiONPs aggravated asthma development, which is closely related to inflammasome activation. Our results provide useful information about the toxicological effects of SiONPs on asthma exacerbation and suggest the need to avoid SiONP exposure especially in individuals with respiratory diseases.

Bruceine D induces lung cancer cell apoptosis and autophagy via the ROS/MAPK signaling pathway in vitro and in vivo

Worldwide, lung cancer remains a leading cause of cancer mortality. Bruceine D (BD) has been shown to induce pancreatic cancer cell death via several different mechanisms. In this study, we demonstrated that BD inhibited lung cancer cell proliferation. Apoptosis and autophagy were the most important mechanisms involved in BD-induced lung cancer cell death, and complete autophagic flux was observed in A549 and NCI-H292 cells. In addition, BD significantly improved intracellular reactive oxygen species (ROS) levels. BD-mediated cell apoptosis and autophagy were almost inhibited in cells pretreated with N-acetylcysteine (NAC), an ROS scavenger. Furthermore, MAPK signaling pathway activation contributed to BD-induced cell proliferation inhibition and NAC could eliminate p-ERK and p-JNK upregulation. Finally, an in vivo study indicated that BD inhibited the growth of lung cancer xenografts. Overall, BD is a promising candidate for the treatment of lung cancer owing to its multiple mechanisms and low toxicity.

Two new compounds with Nrf2 inducing activity from Glycyrrhiza uralensis

Two new compounds including a new 3-arylcoumarin liquiritcoumarin (1), and a new flavonoid glycoside crotoliquiritin (2), together with twelve known compounds (3–14) were isolated from the radix and rhizome of Glycyrrhiza uralensis. Their structures were elucidated on the basis of extensive spectroscopic data analyses. All of the compounds were evaluated for the cytotoxic activities. Only compound 5 showed moderate effects with IC50 of 11.46 μM for A549 cells and IC50 of 7.38 μM for NCI-H292 cells. Compounds 1 and 2 were demonstrated to be Nrf2 pathway activators by using a stable antioxidant response element (ARE)-dependent reporter gene assay together with immunoblot and immunofluorescence analysis.

Ultrasound-assisted synthesis of a new nanostructured Ca(II)-MOF as 5-FU delivery system to inhibit human lung cancer cell proliferation, migration, invasion and induce cell apoptosis

A new porous metal-organic framework material, {Ca3(TATAB)2 (H2O)(MeOH)}(DMF)3}n (1, TATAB3- = 4,4′,4′′-s-triazine-1,3,5-triyltri-p-aminobenzoate), with considerable biocompatibility was synthesized by reaction of the Ca(NO3)2 and the H3TATAB ligand in DMF/MeOH solution. Nanostructure 1 can also be prepared by sonochemical process at ambient temperature. Nitrogen adsorption measurements revealed the presence of micropores as well as moderately high BET surface areas in the activated nanostructured 1 (1a). The incorporation of the drug 5-fluorouracil (5-FU) into 1a was 38.6 wt% per gram of 1a. 5-FU is released in a highly controlled and progressive fashion with 88.7% of the drug released after 72 h. The CCK8 assay was performed to evaluate the inhibitory effect of 5-FU@1a on NCI-H292 and NCI-H460 human lung cancer cells, and the transwell assay was conducted to observe the migration and invasion ability of cancer cells after 5-FU@1a treatment. The Annexin V-FITC/PI assay was carried out to evaluate the level of cancer cell apoptosis. The results above indicate 5-FU@1a has excellent anticancer activity in vitro. The in vivo xenograft model was applied, and the results suggested 5-FU@1a could reduce the volume and the weight of the tumor in vivo.

LncRNA Metastasis-Associated Lung Adenocarcinoma Transcript 1 Promotes Proliferation and Invasion of Non-Small Cell Lung Cancer Cells via Down-Regulating miR-202 Expression.

Objective Accumulating evidences indicate that long non-coding RNAs (lncRNAs) play key roles in cancer. This study aims to clarify role of the metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) in non-small cell lung cancer (NSCLC) and uncover the underlying mechanisms. Materials and Methods In this experimental study, MALAT1 and miR-202 expression in tissues and cell lines were detected using quantitative real time polymerase chain reaction (qRT-PCR) assay. Cell transfection was conducted using Lipofectamine 3000. Cell proliferation was determined with CCK-8 assay. MMP2 and MMP9 expressions were measured with Western blot. Cell invasive ability was evaluated by Transwell assay. Starbase 2.0 tool was used to predict targets of MALAT1. Dual luciferase reporter assay, RNA-binding protein immunoprecipitation assay and RNA pull-down assay were conducted to confirm the potential direct interaction between MALAT1 and miR-202. Results MALAT1 was overexpressed in NSCLC samples and cell lines. High expression of MALAT1 was related to large tumor size (>3 cm), poor histological grade, advanced cancer and tumor metastasis in NSCLC. In vitro assays exhibited that knockdown of MALAT1 remarkably decreased A549 cell growth and invasion capacity, while overexpression of MALAT1 significantly enhanced NCI-H292 cell proliferation and invasion ability. Next, we verified that MALAT1 could act as a competing endogenous RNA (ceRNA) by sponging miR-202 in NSCLC and there is a negative correlation between MALAT1 and miR-202. Besides, overexpression of miR-202 inhibited cell proliferation and invasive ability in MALAT1-overexpressed cells. ConclusionThis study demonstrated that lncRNA-MALAT1 gets involved in NSCLC progression by targeting miR- 202, indicating that MALAT1 may serve as a novel therapeutic target for NSCLC treatment.

Biological Evaluation of Arylsemicarbazone Derivatives as Potential Anticancer Agents.

Fourteen arylsemicarbazone derivatives were synthesized and evaluated in order to find agents with potential anticancer activity. Cytotoxic screening was performed against K562, HL-60, MOLT-4, HEp-2, NCI-H292, HT-29 and MCF-7 tumor cell lines. Compounds 3c and 4a were active against the tested cancer cell lines, being more cytotoxic for the HL-60 cell line with IC50 values of 13.08 μM and 11.38 μM, respectively. Regarding the protein kinase inhibition assay, 3c inhibited seven different kinases and 4a strongly inhibited the CK1δ/ε kinase. The studied kinases are involved in several cellular functions such as proliferation, migration, cell death and cell cycle progression. Additional analysis by flow cytometry revealed that 3c and 4a caused depolarization of the mitochondrial membrane, suggesting apoptosis mediated by the intrinsic pathway. Compound 3c induced arrest in G1 phase of the cell cycle on HL-60 cells, and in the annexin V assay approximately 50% of cells were in apoptosis at the highest concentration tested (26 μM). Compound 4a inhibited cell cycle by accumulation of abnormal postmitotic cells at G1 phase and induced DNA fragmentation at the highest concentration (22 μM).

Pro-inflammatory effects of extracellular Hsp70 on NCI-H292 human bronchial epithelial cell line.

Extracellular Hsp70 (eHsp70) exerts its biological actions via Toll-like receptors 2 and 4, and is increased in sera of chronic obstructive pulmonary disease (COPD) patients. The aim of this study was to explore the pro-inflammatory effects and cytotoxicity of eHsp70 alone and in combination with bacterial components lipoteichoic acid (LTA) and lipopolysaccharide (LPS) on NCI-H292 airway epithelial cells. NCI-H292 cells were treated with recombinant human Hsp70 protein (rhHsp70), LPS, LTA and their combinations for 4, 12, 24 and 48hours. IL-6, IL-8 and TNF-α levels were measured by an ELISA method. Cell viability was determined by the MTS method, and caspase-3/7, caspase-8 and caspase-9 assays. rhHsp70 induced secretion of IL-6 and IL-8 in a concentration- and time-dependent manner, with the highest secretion at 24hours. rhHsp70 combined with LTA had antagonistic and with LPS synergistic effect on IL-6 secretion, while the interactions between rhHsp70 and LPS or LTA on IL-8 were synergistic. TNF-α was not detected in the applied conditions. rhHsp70, LPS or LTA did not affect cell viability, and rhHsp70 even suppressed caspase-3/7 activities. We suggest that pro-inflammatory effects of eHsp70, together with other damaging molecules and/or COPD risk factors, might contribute to the aggravation of chronic inflammation in human bronchial epithelium.