Synthesis of CEG-AgNPs as a Promising MMPs/COX-2/Bcl-2 Signaling Pathway Modulator in BaP-Induced Lung Carcinoma

Cucurbitacins are a class of highly oxidized tetracyclic triterpenoids. It’s hydrophobic properties and poor solubility in water, polymeric micellar systems exhibited improved antitumor efficacy because of a better solubilization and targeting after local and/or systemic administration. The aim of the present work was to evaluate the anticancer activity of CEG-AgNPs against benzo[a]pyren (BaP)-induced lung carcinoma. CEG-AgNPs was prepared, characterized and evaluated for its cytotoxic activity against A549 lung carcinoma cell line. Also, the anticancer activity of CEG-AgNPs (70.25 mg/kg) against BaP-induced lung carcinoma was evaluated in vivo, using 30 adult mice for 43 days. IC50 of CEG-AgNPs against A549 lung carcinoma cell line were approximately 94.47 μg/mL. Administration of BaP (50 mg/kg b.w.) to mice induced lung carcinoma with a significant increase in lung MMP-2, MMP-9, MMP-12, MDA, IL-6 and NF-κB as well as significant decreased in lung CAT, GPx and GSH level. Also, treatment with BaP produced significant increase in lung VEGF-C, COX-2 and Bcl-2 gene expression as compared to control group. Daily oral administration of CEG-AgNPs to mice treated with BaP showed a significant protection against-induced increase in lung MMP-2, MMP-9, MMP-12, MDA, IL-6 and NF-κB levels. The treatment also resulted in a significant increase in lung CAT, GPx and GSH level. In addition, the CEG-AgNPs could inhibit lung VEGF-C, COX-2 and Bcl-2 gene expression as compared to BaP treated mice. The histological and MRI examination showed that a significant normalization has been observed through in CEG-AgNPs treated mice. The biochemical, histological and MRI results showed that CEG-AgNPs have potent anticancer activity against BaP-induced lung carcinoma through modulating multiple cellular behaviours and signaling pathways leading to the suppression of adaptive immune responses.

Toxicity Evaluation of Choline Ionic Liquid-Based Nanocarriers of Pharmaceutical Agent For Lung Treatment With Extra Antitumor Activity

In vitro cytotoxicity evaluation of linear copolymer (LC) containing choline ionic liquid units and its conjugates with an antibacterial drug in anionic form, i.e. p-aminosalicylate (LC_PAS), clavulanate (LC_CLV), or piperacillin (LC_PIP) was carried out. These systems were tested against normal: human bronchial epithelial cells (BEAS-2B), and cancers: adenocarcinoma human alveolar basal epithelial cells (A549), and human non-small cell lung carcinoma cell line (H1299). Cells viability, after linear copolymer LC and their conjugates addition for 72 hours, was measured at concentrations range of 3.125‑100 μg/mL. The MTT test allowed the designation of IC50 index, which was higher for BEAS-2B, and significantly lower in the case of cancer cell lines. The cytometric analyzes, i.e. Annexin-V FITC apoptosis assay and cell cycle analysis as well as gene expression measurements for interleukins IL6 and IL8 were carried out, and showed pro-inflammatory activity of tested compounds towards cancer cells, while it was not observed against normal cell line. In summary, the anticancer property of linear copolymers with antibacterial drugs was shown.

ANTICANCER ACTIVITY OF NATURAL COMPOUND FROM LEMON GRASS LEAVES EXTRACT ON LUNG CARCINOMA CELL LINE A549: AN IN VITRO STUDY

Lemon grass is a widely cultivated plant, whose extracts are known to possess anti-cancer properties. However, studies related to the effect of different solvent extracts of lemon grass on lung cancer cell lines are scarce. This study was conducted to study the effect of various extracts on the viability of the A549 lung carcinoma cell line. Four solvents (hexane, chloroform, ethyl acetate and ethanol) were used for extraction of lemon grass. The effect of these solvent extracts on A549 cell line was studied. Cell viability was studied by 3-(4,5-Dimethylthiazol-2-Yl)-2,5-Diphenyltetrazolium Bromide (MTT) method. The effect of apoptosis on exposure to aforementioned extracts was observed with help of nuclear staining assays and western blotting techniques. Apoptosis was confirmed by nuclear staining using acridine orange ethidium bromide (Ao/EtBr), while pro- and anti-apoptotic cells expressions analysis was conducted using western blotting. All extracts showed cytotoxicity on lung carcinoma cell line, wherein, ethyl acetate and hexane extracts showed up to 48.6±3.8% and 51.7±1.7% viability at 250µg/mL concentration. Up regulation of pro-apoptotic gene expressions like Caspase-3 and inhibition of anti-apoptotic gene expression like Bcl-2 was observed after 24h. An inhibitory concentration (IC50) of 220.01 µg/mL was obtained for the ethyl acetate extract. When observed under fluorescence microscope, stained cells showed an orange red color in the nuclei, indicating the ethyl acetate extract had induced apoptosis potentially for 24 h exposure cells expression, when compared with control cells. The results show that the ethyl acetate extracts were efficient in inhibiting the A549 cell line under in vitro condition, suggesting this extract can be further used as a potential chemotherapy drug.

Adsorption of cisplatin on oxidized graphene nanoribbons for improving the uptake in non-small cell lung carcinoma cell line A549

Background : Graphene nanoribbons are nanosized strips of graphene with unique physicochemical properties like higher drug loading capacity and affinity for tumor cells. Objective: The principal objective of this research was to develop oxidized graphene nanoribbons (O-GNRs)-a based delivery system for cisplatin against non-small cell lung carcinoma cell line A549 by selective endocytosis. Method: The O-GNRs prepared using various synthetic steps like oxidative unzipping were evaluated for various parameters like morphology, Fourier Transform Infrared (FTIR) study, % adsorption efficacy, Differential scanning colometric (DSC) study, and in-vitro efficacy studies. Results: Graphene nanoribbons with the length of 200-250 nm and width of 20-40 nm were obtained. The FTIR spectrum of drug-loaded O-GNRs exhibited a characteristic peak at 1550 cm-1 (-N-H group) of cisplatin. The DSC indicated the presence of sharp endothermic peaks at 59 ºC (PEG), 254 ºC (-C-NH3) and 308.6 ºC (-C-Pt). The % adsorption efficiency was found to be 74.56 ± 0.798% with in-vitro release in controlled manner (63.36 % ± 0.489 %) for 24 h. Conclusion: The nanoformulation showed an average inhibition of 22.72% at a lower dose of cisplatin (> 25%) by passive targeting cell line A549 by DNA alkylation. In the near future, graphene-based systems will establish potential nanosystems in cancer treatment due to the additive effect of graphene with various therapeutic agents.

Cytosolic 5′-Nucleotidase II Is a Sensor of Energy Charge and Oxidative Stress: A Possible Function as Metabolic Regulator

Cytosolic 5′-nucleotidase II (NT5C2) is a highly regulated enzyme involved in the maintenance of intracellular purine and the pyrimidine compound pool. It dephosphorylates mainly IMP and GMP but is also active on AMP. This enzyme is highly expressed in tumors, and its activity correlates with a high rate of proliferation. In this paper, we show that the recombinant purified NT5C2, in the presence of a physiological concentration of the inhibitor inorganic phosphate, is very sensitive to changes in the adenylate energy charge, especially from 0.4 to 0.9. The enzyme appears to be very sensitive to pro-oxidant conditions; in this regard, the possible involvement of a disulphide bridge (C175-C547) was investigated by using a C547A mutant NT5C2. Two cultured cell models were used to further assess the sensitivity of the enzyme to oxidative stress conditions. NT5C2, differently from other enzyme activities, was inactivated and not rescued by dithiothreitol in a astrocytoma cell line (ADF) incubated with hydrogen peroxide. The incubation of a human lung carcinoma cell line (A549) with 2-deoxyglucose lowered the cell energy charge and impaired the interaction of NT5C2 with the ice protease-activating factor (IPAF), a protein involved in innate immunity and inflammation.

Synthesis, Biological Profile, and Molecular Docking of Some New BisImidazole Fused Templates and Investigation of Their Cytotoxic Potential as Anti-tubercular and/or Anticancer Prototypes

Background: There is a great need to discover more drugs with antimycobacterial activities to fight lung cancer and tuberculosis (two of the deadliest diseases world-wide). To our knowledge, the present study is the first to report antimycobacterial activity of imidazole-fused heterocycles. Objective: Construction of some bis-imidazole fused heterocycles with potential anti-tubercular and/or potent antitumor activities. Method: A series of bis-imidazole fused derivatives 6-8 and 13-16 was constructed using bis-phenacyl bromide derivative 2 as a synthetic platform. Compound 2 was also used to access bis-quinoxaline 20, bis-benzothiazine derivatives 23, and bisthiazolopyrimidine derivatives 26. The new bis-imidazole derivatives were evaluated for their anticancer activity against lung carcinoma cell line (A-549) using Cisplatin as a reference drug. The new compounds were also screened for their antitubercular activity against M. tuberculosis (ATCC 25177) using Isoniazid as a reference drug. Result: Among the new bis-imidazole derivatives, three examples showed remarkable antitumor activities while five other compounds showed high antimycobacterial activity. Conclusion: A novel series of bis-imidazole fused heterocycles was developed. Multiple prototypes of this new series showed remarkable anti-tubercular and/or potent antitumor activities.

Epigenetic Modifications, and Alterations in Cell Cycle and Apoptosis Pathway in A549 Lung Carcinoma Cell Line upon Exposure to Perfluoroalkyl Substances

Per- and polyfluoroalkyl substances (PFAS) are a group of human-made compounds with strong C-F bonds, and have been used in various manufacturing industries for decades. PFAS have been reported to deleterious effect on human health, which has led to studies identifying the possible toxicity and toxicity routes of these compounds. We report that these compounds have the potential to cause epigenetic modifications, and to induce dysregulation in the cell proliferation cycle as well as apoptosis in A549 lung cancer cells when exposed to 10-, 200- and 400 μM concentrations of each compound. Our studies show that exposure to perfluorooctanoic acid (PFOA) and perfluorooctane sulfonic acid (PFOS) may cause hypomethylation in the epigenome, but changes in the epigenetic makeup are not evident upon exposure to GenX. We establish that exposure to lower doses of these compounds causes the cells’ balance to shift to cell proliferation, whereas exposure to higher concentrations shifts the balance more towards apoptosis. Furthermore, the apoptosis pathway upon exposure to GenX, PFOA, and PFOS has also been identified. Our findings suggest that exposure to any of these compounds may have profound effects in patients with pre-existing lung conditions or could trigger lung cancinogenesis.

Short-Chain Hydroxyl CoA Dehydrogenase Probably is the Central Player in DNA Replication and Glucose Consumption in Human Non-Small Lung Carcinoma Cell Line

: Hydroxyl CoA Dehydrogenase (HADH) is one of the key enzymes in fatty acid β-oxidation. Recently, Hydroxyl CoA Dehydrogenase gene mutation and knockdown were found to be correlated with hyperinsulinemia and central nervous system diseases. As the HADH is one of the critical enzymes in the β-oxidation pathway, the interconnection between HADH and tumorigenicity still is unclear. So, we used Short hairpin RNA (ShRNA) to knock down short-chain hydroxyl CoA dehydrogenase (HADHSC) in human non-small lung carcinoma cell line, H1299, followed by checking cell proliferation, DNA replication, and mRNA level of some the most essential enzymes in glycolysis cycle and Krebs. Cell proliferation was checked by comparing the cell numbers in knockdown and control cells. DNA replication in the H1299 cell line was studied after applying 5-ethynyl 2’-deoxyuridine (EDU) and 4’-6 diamidino-2-phenylindol (DAPI) DNA synthesis Assay. The data revealed a significant decrease in cell proliferation and DNA replication in the cells that the HADHSC was knocked down compared to the control cells. Besides, mRNA levels of the enzymes that needed adenosine triphosphate (ATP) for their activity were decreased abruptly. Furthermore, lactate dehydrogenase (LDHA) mRNA level decreased, and glucose uptake assay showed a tremendous decrease in glucose consumption by H1299 cells with HADHSC knockdown.