Human Cancer Cell Line A549 Research Articles

Design, synthesis, crystal structure, molecular docking studies of some diorganotin(IV) complexes derived from the piperonylic hydrazide Schiff base ligands as cytotoxic agents

Inspired by the role of organotin(IV) complexes in inhibition of cancer cell growth and interaction with different target proteins, we have synthesized a series of new organotin(IV) complexes of Schiff base ligands of benzylidene benzohydrazide analogues. The structural elucidation of compounds was done by spectroscopic studies showing tridentate nature (NOO) of Schiff base ligands having pentacoordinated geometry around the central tin metal. The X-ray crystallographic study of complex 9 (Me2SnL2) revealed the distorted trigonal bipyramidal geometry (SnO2NC2). The cytotoxic activity of compounds was tested against human cancer cell lines A549, Hela, MCF7 and normal cell line L6 using MTT assay. Compound 12 (Ph2SnL2), 14 (Et2SnL3), 19 (Bu2SnL4) was found most active against tested cell lines having IC50 value 22.909–32.303 μM. Further, molecular docking study was performed for the active compounds 12, 14 and 20 at 3D space of enzymes Tyrosine kinase and EGFR kinase domain.

Microbial Transformation of Broussochalcones A and B by Aspergillus niger.

Broussochalcones A (BCA, 1) and B (BCB, 2) are major bioactive constituents isolated from Broussonetia papyrifera, a polyphenol-rich plant belonging to the family Moraceae. Due to their low yields from natural sources, BCA (1) and BCB (2) were prepared synthetically by employing Claisen-Schmidt condensation, and these were used as substrates for microbial transformation to obtain novel derivatives. Microbial transformation of BCA (1) and BCB (2) with the endophytic fungus Aspergillus niger KCCM 60332 yielded 10 previously undescribed chalcones (1a-1e and 2a-2e). Their structures were established based on the spectroscopic methods. The cytotoxicity of BCA (1), BCB (2), and their metabolites (1a-1e and 2a-2e) was determined by human cancer cell lines A375P, A549, HT-29, MCF-7, and HepG2, with 1e shown to be most cytotoxic.

Inonotus obliquus Polysaccharides Inhibited Cellular Growth of NCI-H23 and A549 Lung Cancer Cells Through G0/G1 Cell Cycle Arrest and ROS Mediated Cell Death

Chaga mushroom (Inonotus obliquus) has been used for a long time as a folk medicine for treating multiple diseases in several parts of the world without rendering any undesired toxicity. In this study, I. obliquus polysaccharides (IOP) were extracted and assessed to determine their anti-tumorigenic potential in human lung cancer cell lines NCI-H23 and A549 using cytotoxicity and apoptosis assays. MTT assay revealed a significant reduction in the cell viability (p < 0.05) for NCI-H23 and A549 cell lines exposed to IOP (5-200 µg/mL) in a concentration-dependent manner with IC50 of 100 µg/mL for both cell lines. Cell lines exposed to 50 and 100 µg/mL of IOP were further analyzed. IOP arrested the cancer cell growth at G0/G1 stage that can further implicate an anti-proliferative effect in cancer cells. Intracellular reactive oxygen species (ROS) generation was detected using DCFH-DA dye demonstrated increased levels of ROS generation (p < 0.05). Assessment of mitochondrial membrane potential using JC-1 dye exhibited decreased membrane potential, characterized by the low dye-intake, as shown by flow cytometry. In addition, Annexin-V/FITC analysis using flow cytometry demonstrated a significantly increased number of apoptotic cells (p < 0.05) in both cancer cell lines in a concentration-dependent manner. These results showed that IOP can induce apoptosis in both tumor cell lines, and therefore might be considered as an effective anti-tumor agent that could be further exploited in clinical settings.

Network pharmacology and molecular docking reveal the mechanism of Scopoletin against non-small cell lung cancer

AimsScopoletin is a natural anticarcinogenic and antiviral coumarin component. Many studies have proved its anti-cancer effect, and after the preliminary screening of this study, Scopoletin had the best inhibitory effect on Non-small cell lung cancer (NSCLC). But its mechanism for treating NSCLC is still unclear. Therefore, network pharmacology and molecular docking technology were used to explore the potential anti-NSCLC targets and pathways of Scopoletin. The results were verified in vitro. Main methodsFirst, Scopoletin was isolated from Fennel and screened to conduct cell proliferation assay on Human lung cancer cell line A549, Human colon cancer cell line HCT-116 and Human hepatoma cell line HepG2 respectively, through the MTT test. Then, the key targets and related pathways were screened through Protein-protein Interaction (PPI) network and “component-target-pathway” (C-TP) network constructed by network pharmacology. And the key targets were selected to dock with Scopoletin via molecular docking. A549 and Human normal lung epithelial cell BEAS-2B were used to verify the results, finally. Key findingsThrough MTT, A549 was chosen as the test cancer cell. From network pharmacology, 16 targets, 27 signaling pathways and 16 GO items were obtained (P < 0.05). The results of PPI network and molecular docking showed that EGFR, BRAF and AKT1 were the key targets of Scopoletin against NSCLC, which were consistent with the western-blot results. SignificanceThrough network pharmacology, molecular docking and experiments in vitro, Scopoletin was verified to against NSCLC through RAS-RAF-MEK-ERK pathway and PI3K/AKT pathway.

Discovery of facile amides-functionalized rhodanine-3-acetic acid derivatives as potential anticancer agents by disrupting microtubule dynamics

Microtubule dynamics are crucial for multiple cell functions, and cancer cells are particularly sensitive to microtubule-modulating agents. Here, we describe the design and synthesis of a series of (Z)-2-(5-benzylidene-4-oxo-2-thioxothiazolidin-3-yl)-N-phenylacetamide derivatives and evaluation of their microtubule-modulating and anticancer activities in vitro. Proliferation assays identified I20 as the most potent of the antiproliferative compounds, with 50% inhibitory concentrations ranging from 7.0 to 20.3 µM with A549, PC-3, and HepG2 human cancer cell lines. Compound I20 also disrupted cancer A549 cell migration in a concentration-dependent manner. Immunofluorescence microscopy, transmission electron microscopy, and tubulin polymerisation assays suggested that compound I20 promoted protofilament assembly. In support of this possibility, computational docking studies revealed a strong interaction between compound I20 and tubulin Arg β369, which is also the binding site for the anticancer drug Taxol. Our results suggest that (Z)-2-(5-benzylidene-4-oxo-2-thioxothiazolidin-3-yl)-N-phenylacetamide derivatives could have utility for the development of microtubule-stabilising therapeutic agents.

Correlation between Plasma Cellular Retinoic Acid-Binding Protein 2 and Proliferation, Migration, and Invasion of Non-Small-Cell Lung Cancer Cells.

Our primary aim of the current study was to explore the correlation between plasma CRABP2 and migration, proliferation and invasion of non-small cell lung cancer (NSCLC) cells. Human lung cancer cell line A549 was used in the present study, which was cultured in 6-well plates (1 × 106 cells/well) and then transfected with pcDNA-CRABP2 and pcDNA, siRNA with the use of Lipofectamine 2000 based on the manufacturer's protocol. The expression of CRABP2 mRNA was detected through real-time PCR. Proliferation was further detected using MTT assays, and apoptosis was monitored and recorded with the application of flow cytometry. The expression of E-cadherin, MMP9, vimentin and related pathway proteins was detected by Western blotting assays. Transwell assays and cell scratch assays were utilized for the detection of migration and invasion ability of A549 cells. RT-PCR results showed The CRABP2 mRNA transcript levels in the CRABP2 overexpression group were higher when comparing those of the empty vector group (P < 0.05). By MTT assays, CRABP2 overexpression promoted cellular proliferation, while CRABP2 downregulation inhibited cellular proliferation. CRABP2 overexpression inhibited cell apoptosis and promoted cellular proliferation. The number of TUNEL staining positive cells was the lowest in the CRABP2 overexpression group, and the siRNA transfection group had increased apoptosis. CRABP2 downregulation reduced EMT in cells and cell migration and invasion reflected from western blotting results and cell migration and invasion assay results, respectively. Inhibition of plasma CRABP2 expression offers the potential in terms of reducing the expression of MAPKs and proteins in the NF-κB pathway and inhibiting the proliferation and migration of NSCLC cells, which is ideally suited for further treatment for NSCLC.

Synthesis, characterization of novel Sesamol substituted with thiazolidin-4-one derivatives and their evaluation for anti-oxidant and anti-cancer activities

In recent years for the development of novel anticancer hybrids with two or more pharmacophores of bioactive scaffolds to produce a single molecule by blending or linking has emerged as a simple strategy by providing an effective control on malignant process with improved biological potential. In this view, Sesamol being an active scaffold with important biological activities was modified and linked with various substitutions to afford thiazolidin-4-one derivatives. Herein, we report the synthesis, characterization of around twenty one compounds and their evaluation for antioxidant and anticancer potentials. The structures of these compounds were established by IR, 1H NMR, 13C NMR, and Mass. The compounds 1–15 were evaluated for in vitro cytotoxicity against three human lung cancer cell lines (A549, MCF-7 and HeLa), respectively. In preliminary MTT [3-(4,5- dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] cytotoxicity studies compounds 9 and 12 were found most effective. In A549 cancer cells the CTC50 (50% of cytotoxicity inhibition) were observed at 84.27 ± 2.7 & 91.98 ± 4.2 µg/ml, respectively. Moreover, in MCF-7 and HeLa cells the CTC50 values were observed at 174.3 ± 4 4.1, 69.15 ± 3.4 and 42.90 ± 3.5, 180.77 ± 4.6 µg/ml, respectively. Compounds 9 and 12 also exhibited best DNA damage activity via cell apoptosis when screened against Human breast cancer cell lines A549 and MCF-7 when compared to standard doxorubicin. In this study, Compounds 1, 9, 11 and 12 (at concentration 100 mg/kg/body wt.) were selected for in vivo anticancer activity against Dalton’s Lymphoma Ascites bearing mice. Among the tested compounds all has shown modest antitumor activity and merited anti-cancer activity due to its cytotoxic properties when compared with standard methotrexate.

Effect of Menadione and Combination of Gemcitabine and Cisplatinon Cancer Stem Cells in Human Non-small Cell Lung Cancer (NSCLC) Cell Line A549.

Non-small cell lung cancer (NSCLC) is the most common type of lung cancer. Chemotherapy-induced adverse effects and resistance of NSCLC to conventional drugs reduce the efficacy of current therapies. Tumors contain a small population of cancer stem cells (CSCs) that play a critical role in tumor initiation, maintenance, and drug resistance that finally lead to cancer recurrence. Therefore, CSC-targeting therapies can offer the best hope for developing curative cancer therapies. Vitamins have a high potential for cancer prevention and treatment. Vitamins also ameliorate the side effects which occur in chemo-radio therapy. Menadione (2-methyl-1,4-naphthoquinone/vitamin-K3) is a synthetic form of vitamin K that indicated antitumor activities. The purpose of this study was to evaluate the anti-CSCs effect of menadione and combination of cisplatin and gemcitabine as a first-line treatment in patients with NSCLC on the NSCLC cell line A549. MTT results displayed decreased cell survival after treatment with cisplatin/gemcitabine for 48 h treatment (IC50 values 0.25 µM for cisplatin and 5 µM for gemcitabine). Menadione also inhibited the cell growth in A549 cells (IC50: 16 µM). Quantitative RT-PCR showed significant downregulation of CSC markers (Oct4, Nanog, Sox2, Aldh1, Abcb1, CD44, and CD133) and Snail, epithelial-mesenchymal transition marker, after treatment with menadione and cisplatin/gemcitabine. Flow cytometry showed CD44-positive cells that constitute a high percentage (70%) of A549 cells reduced significantly after treatment with cisplatin/gemcitabine or menadione. However, A549 cells did not show a significant population positive for CD133 and ABCB1 (less than 0.05%), and these fractions did not change after treatment with two agents.

Inhibition of metastasis and suppression of pERK1/2 and pFAK expression by Solanum xanthocarpum crude extracts in human lung cancer cell line A549 in vitro

Solanum xanthocarpum is an important part of natural remedies for lung in Ayurveda. Lung cancer metastasis is a major challenge in the present time. Expressions and activation of focal adhesion kinase (FAK) and extracellular receptor kinases 1 and 2 (ERK1/2) are key checkpoints in cancer which lead to an invasion, epithelial-mesenchymal transition (EMT) and metastatic progression. In this study, the efficacy of the whole plant and root crude extracts prepared in ethyl acetate and chloroform on highly metastatic lung cancer cell line A549 was evaluated. The effect of crude extracts on cell migration was scrutinized using transwell migration assessment. The phosphorylated forms of proteins, pFAK and pERK1/2 levels, were estimated using ELISA. Zymography technique was utilized to evaluate the activities of MMP-2 and MMP-9 proteins. The EMT markers, vimentin and N-cadherin gene expressions were analyzed using semi-quantitative RT-PCR. The extracts significantly reduced single-cell migration. The expression levels of pFAK and pERK1/2 notably decreased and FAK and ERK1/2 remained unchanged. Moreover, gelatinase activity, vimentin and N-cadherin expression were also significantly reduced in treated cells compared to untreated cells. The crude extracts of S. xanthocarpum impaired the metastatic progression of A549 cells in vitro.

Structure and absolute configuration assignments of ochracines F-L, chamigrane and cadinane sesquiterpenes from the basidiomycete Steccherinum ochraceum HFG119.

Ochracines F–L (1–7), seven previously undescribed chamigrane and cadinane sesquiterpenoids, together with four known chamigranes were isolated from cultures of the wood-decaying fungus Steccherinum ochraceum HFG119. Ochracines F–L were structurally characterized by extensive analysis of HRMS and NMR spectroscopic data. The relative configurations were assigned through a combination of NOE correlations and J-based configuration analysis (JBCA), while the absolute configurations were determined by X-ray single-crystal diffraction, and calculated methods (ECD, [α], 13C NMR). All the new isolates were evaluated for their cytotoxicity against five human cancer cell lines HL-60, SMMC-7721, A549, MCF-7, and SW-480, and inhibitory activity on NO production in RAW 264.7 macrophages.

Sulfated magnesium zirconate catalyzed synthesis, antimicrobial, antioxidant, anti-inflammatory, and anticancer activity of benzo[d]thiazole-hydrazone analogues and its molecular docking

A comprehensive demonstration of novel catalytic synthetic pathway using sulfated zirconia (solid acid catalyst) to generate a library of bioactive compounds precisely, the benzo[d]thiazole-hydrazone derivatives (4a-n). The compounds are characterized using ubiquitous spectroscopic techniques such as, IR, 1H and 13C NMR, HR-Mass. The set of molecules are screened in vitro biological activities namely, antimicrobial, antioxidant, anti-inflammatory and anticancer which are aptly supported by the molecular docking studies. Among the compounds, 4a and 4e are observed to be the most potent antibacterial agents where as 4a and 4c displayed significant antifungal activity. On the contrary, compounds 4e and 4j showed good antioxidant properties and 4b and 4j exhibited excellent anti-inflammatory activity. The compound 4d found to be a potent anti-cancer agent against tested human cancer cell lines MIAPaca2, HeLa, A549, and HCT116 (IC50 values of 19.09 ± 0.50, 19.93 ± 0.25, 7.76 ± 0.50, and 5.05 ± 0.25 μM respectively). Molecular docking studies were performed using DNA Gyrase, N-myristoyltransferase, COX-2, EGFR, HER2, and VEGFR2 proteins. Notably, compound 4d strongly binds to receptors EGFR and HER2 as inferred by their binding energies compared to standard inhibitors Gefitinib (EFGR), Lapatinib (EGFR), Afatinib (HER2), and Canertinib (HER2).

Preclinical in vitro screening of newly synthesised amidino substituted benzimidazoles and benzothiazoles

Newly synthesised benzimidazole/benzotiazole derivatives bearing amidino, namely 3,4,5,6-tetrahydropyrimidin-1-ium chloride, substituents have been evaluated for their potential antitumor activity in vitro. Compounds and standard drugs (doxorubicin, staurosporine and vandetanib) were tested on three human lung cancer cell lines A549, HCC827 and NCI-H358. We tested compounds in MTS citotoxicity assay and in BrdU proliferative assay performed on 2 D and 3 D assay format. Because benzmidazole scaffold is similar to natural purines, we tested the most active compounds for ability to induce cell apoptosis of A549 by binding to DNA in comparison with doxorubicin and saturosporine. Additionally, the ADME properties of the most active benzothiazole/benzimidazole and non-active compounds were determined to see if the different ADME properties are the cause of different activity in 2 D and 3 D assays, as well as to see if the tested active compounds have drug like properties and potency for further profilation. ADME characterisation included solubility, lipophilicity, permeability, metabolic stability and binding to plasma proteins. In general, the benzothiazole derivatives were more active in comparison to their benzimidazole analogues. The exception was 2-phenyl substituted benzimidazole 6a being active with very pronounced activity especially towards HCC827 cells. All active compounds have similar mode of action on A549 cell line as standard compound doxorubicin, which binds to nucleic acids with the DNA double helix. Tested active benzothiazole compounds were characterised by moderate to good solubility, good metabolic stability, low permeability and high binding to plasma proteins. One tested active benzimidazole derivative showed ADME properties, but lower lipophilicity resulted in low PPB and higher metabolic instability. In addition, no significant difference was observed in ADME profile between active and non-active compounds.

Cytotoxic Effects of Flubendazole in Human Lung Cancer Cell Line A549

Background: The development of effective anticancer drugs is a significant health issue. Previous studies showed that members of the benzimidazole family have anticancer effects on several cancers Objectives: The present study investigated the cytotoxic effect of flubendazole on A549 human lung cancer cells. Methods: The A549 cells were treated with flubendazole at 1, 2, 5, and 10 µM concentrations for three days. Cell viability was measured by the MTT assay and Acridine orange staining. Also, the expressions of P62 and Beclin -1 were analyzed by qRT-PCR analysis. Results: Cell viability of A549 cells, in a concentration-dependent manner, showed significant differences between the treatment and control groups, and the IC50 value was determined to be 2 µM. Also, flubendazole reduced the expression of P62 and increased the expression of Beclin 1 in treated cells. Conclusions: Flubendazole induces cell death in A549 cells in a dose and time-dependent manner and can offer new factors in lung cancer therapeutic strategies.

Self-Assembled Disulfide Bond Bearing Paclitaxel-Camptothecin Prodrug Nanoparticle for Lung Cancer Therapy.

Self-assembled prodrugs (SAPDs), which combine prodrug strategy and the merits of self-assembly, not only represent an appealing type of therapeutics, enabling the spontaneous organization of supramolecular nanocomposites with defined structures in aqueous environments, but also provide a new method to formulate existing drugs for more favorable outcomes. To increase drug loading and combination therapy, we covalently conjugated paclitaxel (PTX) and camptothecin (CPT) through a disulfide linker into a prodrug, designated PTX-S-S-CPT. The successful production of PTX-S-S-CPT prodrug was confirmed by nuclear magnetic resonance (NMR) and high-resolution mass spectrometry (HRMS). This prodrug spontaneously undergoes precipitation in aqueous surroundings. Taking advantage of a flow-focusing microfluidics platform, the prodrug nanoparticles (NPs) have good monodispersity, with good reproducibility and high yield. The as-prepared prodrug NPs were characterized with dynamic light scattering (DLS) and transmission electron microscopy (TEM), demonstrating spherical morphology of around 200 nm in size. In the end, the self-assembled NPs were added to mouse embryonic fibroblast (MEF), mouse lung adenocarcinoma and Lewis lung carcinoma (LLC) cell lines, and human non-small cell lung cancer cell line A549 to evaluate cell viability and toxicity. Due to the redox response with a disulfide bond, the PTX-S-S-CPT prodrug NPs significantly inhibited cancer cell growth, but had no obvious toxicity to healthy cells. This prodrug strategy is promising for co-delivery of PTX and CPT for lung cancer treatment, with reduced side effects on healthy cells.

Antimetastatic effects of VOflavonoid complexes on A549 cell line

BackgroundNon-small-cell lung cancer (NSCLC) is the most frequent type of lung cancer and more than 90 % of mortality is due to metastasis-related deaths. Flavonoids are considered nutraceuticals due to the variety of pharmacological properties. In this paper, we studied the effects of baicalin, silibinin, apigenin, luteolin, and its oxidovanadium(IV) cation complexes on the viability, adhesion to fibronectin, invasion, and migration on human lung cancer cell line A549. In addition, in order to complete the study of the interaction of VOflavonoids and bovine serum albumin (BSA), the binding ability of silibinin and VOsil to the protein was evaluated. MethodTo establish the non-cytotoxic concentration range of the tested compounds, the cancer cell viability was evaluated using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay. Cell migration and invasion assays were performed using Boyden chambers and adhesion assay using MTT method. The interaction of compounds with BSA were investigated in physiological buffer (pH = 7.4) by fluorescence spectroscopy. ResultsAll complexes inhibited the metastatic cascade steps to a greater extent than their respective ligands. Likewise, based on binding constant values (Kb) for BSA-silibinin and BSA-VOsil, we can suggest that both compounds can interact with the protein. ConclusionAlthough all the complexes suppressed cell adhesion, invasion and migration, VOlut can be considered as a good candidate to continue the trials because it presented encouraging results as a potential antitumor and antimetastatic agent, and can be transported by BSA.

Free Radical Scavenging and In vitro Cytotoxic Activity of Bugnay (Antidesma bunius) Leaves Extract against A549 Human Lung Adenocarcinoma and HCT-116 Human Colorectal Cancer Cell Lines

Cancer is one of the significant causes of mortality worldwide. Studies on antineoplastic drugs focused on natural products have revealed several mechanisms to inhibit cancer cells. Bugnay (Antidesma bunius) leaves showed potentials due to its activity observed against brine shrimp and breast cancer cells. However, there is still limited knowledge about its activity against other human cancer cells. This study focused on determining the phytochemical compounds in A. bunius leaves extract, the free radical scavenging activity of the extract using the Diphenylpicrylhydrazyl (DPPH) method, and in vitro cytotoxic activity against two cancer cell lines, namely HCT-116 human colorectal and A549 human lung adenocarcinoma cancer cell lines by using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The phytochemicals identified were unsaturated lactones, flavonoids, phenolics, diterpenes, saponins, tannins, carbohydrates, and reducing sugars. The extract showed significant free radical scavenging activity and a direct correlation of activity with concentration levels. It also exhibited cytotoxic activity against HCT-116 human colorectal and A549 human lung adenocarcinoma. Hence, A. bunius leaves have the potential to be a source of antioxidant and antineoplastic compounds. This warrant further isolation of the compounds for chemotherapeutic purposes.Keywords: Antidesma bunius, Bugnay, Cancer, Cytotoxicity, Radical

Upper rim modified calix[4]arene towards selective turn-on fluorescence sensor for spectroscopically silent metal ions

This work describes the synthesis of a novel sensor calix[4]arene thiosemicarbazone (Lig), consisting of a calix[4]arene thiosemicarbazone moiety capable of detecting Cu2+, Hg2+, Zn2+ colorimetrically and for the selective detection of Zn2+ by “Turn On” fluorescence change in DMSO-H2O (9:1, v/v, pH = 7.2) solution amongst various toxic heavy metal ions. The sensor shows about 240-fold fluorescence enhancements at 474 nm (Stokes shift-140 nm) when bound to a Zn2+ ion with a detection limit of about 15 µM for the Zn2+ ion. Such fluorescence could be visualized by normal unaided eyes offering a simple, visual detection process. DFT theoretical study has been used to further support the high efficiency of the sensor. The binding ability of the Lig with Zn2+ ions has been used to evaluate the biofunctional properties of the synthesized ligand. The synthesized Lig is found to possess the cytotoxic activity and a better penetrating ability towards the A549 human lung cancer cell line.

Chromenopyridin A, a new N-methoxy-1-pyridone alkaloid from the endophytic fungus Penicillium nothofagi P-6 isolated from the critically endangered conifer Abies beshanzuensis

A new N-methoxy-1-pyridone alkaloid [chromenopyridin A (1)] and four known compounds (2–5) were isolated and identified from the endophytic fungus Penicillium nothofagi P-6, which was derived from the bark of the critically endangered conifer Abies beshanzuensis. Their structures were elucidated by extensive spectroscopic analyses and single-crystal X-ray diffraction. Among the isolates, compound 1 showed considerable cytotoxicities against the A549 and Hela human cancer cell lines, with IC50 values of 14.7 and 11.3 μM. In addition, compounds 1 and 4 exhibited potent antibacterial activity against Staphylococcus aureus with MIC values of 62.5 and 15.6 μg/mL, respectively.

Formulation and evaluation of self-nanoemulsifying drug delivery system of brigatinib: Improvement of solubility, in vitro release, ex-vivo permeation and anticancer activity

The aim of this study was to develop a novel self-nanoemulsifying drug delivery system (SNEDDS) of brigatinib, a poorly soluble anticancer drug, so as to improve its solubility, in vitro release, ex-vivo permeation and anticancer activity. Components of SNEDDS were screened on the basis of maximum solubility of brigatinib. Oleic acid, Tween 20 & diethylene glycol monoethylether were selected as oil phase, emulsifier and co-solvent respectively. Various brigatinib-SNEDDS were developed by spontaneous, low energy, simple mixing method. Developed SNEDDS were optimized on the basis of self-emulsifying capacity, globule size, size distribution, thermodynamic stability and capacity to solubilize maximum amount of brigatinib. The optimized SNEDDS was characterized for physicochemical evaluations such as viscosity, conductivity, pH and refractive index in addition to morphological evaluation of globule size and shape by transmission electron microscopy. The optimized SNEDDS exhibited approximately 205-fold enhancement in the saturation solubility of the brigatinib that resulted in improved in vitro release of drug from SNEDDS as compared to pure drug (P < 0.01). The ex-vivo permeation of brigatinib-SNEDDS across rat intestinal sacs was also improved as compared to pure drug (P < 0.01). The cytotoxic activity of brigatinib-SNEDDS against A549 human lung cancer cell lines during MTT assay were significantly improved as compared to pure brigatinib probably due to improved permeability of the SNEDD loaded brigatinib.

Biomarker discovery in highly invasive lung cancer cell through proteomics approaches.

Lung cancer is one of the leading causes of cancer-related death worldwide. The most common type of lung cancer is non-small cell lung cancer (NSCLC). When NSCLC is detected, patients are typically already in a metastatic stage. Metastasized cancer is a major obstacle of effective treatment and understanding the mechanisms underlying metastasis is critical to treat cancer. Herein, we selected an invasive subpopulation from the human lung cancer cell line A549 using the transwell system and named it as A549-I5. Invasive and migratory activities of this cell line were analysed using wound healing, invasion, and migration assays. In addition, epithelial-mesenchymal transition (EMT) markers, such as Snail 1, Twist, Vimentin, N-cadherin and E-cadherin, were assessed through immunoblotting. In comparison to A549 cells, the invasive A549-I5 lung cancer cells had enhanced invasiveness, motility and EMT marker expression. Proteomic analysis identified 83 significantly differentially expressed proteins in A549-I5 cells. These identified proteins were classified according to their cellular functions and most were involved in cytoskeleton, redox regulation, protein degradation and protein folding. In summary, our results provide potential diagnostic markers and therapeutic candidates for the treatment of NSCLC metastasis. SIGNIFICANCE OF THE STUDY: When NSCLC is detected, most patients are already in a metastatic stage. Herein, we selected an invasive subpopulation from a human lung cancer cell line which had increased EMT markers as well as high wound healing, invasion and migration abilities. Proteomic analysis identified numerous proteins associated with functions in cytoskeleton, redox regulation, protein degradation and protein folding that were differentially expressed in these cells. These results may provide potential diagnostic markers and therapeutic candidates for the treatment of NSCLC metastasis.