Lung Cancer Cell Line A549 Research Articles

Klebsiella pneumoniae Infection in Alveolar Type II Epithelial Cells; Antibiotic-Vitamin Combination Therapeutic Effect

Aim: In this study, it was aimed to establish an infection model with Klebsiella pneumoniae on A549 Lung cancer cell line and to evaluate the effect of additional vitamins on the antibacterial effect of ampicillin sulbactam in the treatment. Cell culture and pathology results were determined in parallel with microbial analysis. Methods: Minimal inhibitory concentration (MIC), fractional inhibitory concentration (FIC), biofilm optical density of ampicillin sulbactam, vitamins E, K1 and P on Klebicella pneumoniae ATCC 700603 strain were determined. Cytotoxic activity on A549 cancer cell line in parallel with microbial analysis and pathology results were determined. Results: Ampicillin Sulbactam MIC dose range used in the treatment of Klebsiella pneumoniae infection is 16 mg/L. Looking at the FIC dose ranges, Vitamin K1+Ampicillin sulbactam, Vitamin P+Ampicillin sulbactam 2.5mg/ml+0.5µg/ml, Vitamin E+Ampicillin sulbactam 5mg/ml+0.5µg/ml, K1+P+E+Ampicillin sulbactam 2,5mg/ml+2.5mg/ml+5mg/ml+0.5µg/ml it was determined that it was. It was determined that it has a synergistic effect in combination with vitamins K1 and P antibiotics. In addition, in the presence of biofilm below 570 nm, the OD values of K1, P, E and K1+P+E+ antibiotic combinations were found to be 1.006, 0.969, 1.096 and 1.015, respectively. As a result of cell culture studies and evaluation of pathology results, it was determined that increasing the dose of Ampicillin sulbactam to 8 µg/ml increased the antibacterial effect and the vitamin combination was more effective. Conclusions: Antibiotic and vitamin combination was found to be more effective in Klebsiella pneumoniae infection in our study. This situation contributes to the search for alternative antimicrobials.

Syntheses and characterization of novel antimony (III) and bismuth (III) derivatives containing β-enamino ester along with antimicrobial evaluation, DFT calculation, and cytotoxic study.

Four novel antimony (III) and bismuth(III) complexes of the kind Cl-Sb-O-C(OR)-CH(CH3 )C-NH-(CH2 )2 -NH-C(CH3 )CH:C(OR)-O [where R = -CH3 , M = Sb (1a); R = -C2 H5 , M = Sb (1b); R = -CH3, M = Bi (1c); R = -C2 H5 , M = Bi (1d)] were successfully prepared by reacting antimony(III)chloride and bismuth(III)chloride with sodium salt of β-enamino esters in 1:1 stoichiometry, which were further structurally characterized by physicochemical and IR, 1 H, 13 C NMR spectral and mass spectrometry. Structural analysis revealed that all four derivatives of both antimony and bismuth display octahedarl geometry which has been optimized through computational studies. These derivatives along with their parent ligands were subsequently assayed in vitro for antibacterial (Bacillus subtilis, Pseudomonas aeruginosa) and antifungal (Aspergillus niger and Candida albicans) activities. Synthesized complexes were more efficacious in terms of biological activities as compared to parent ligands Further synthesized compounds were evaluated for their in vitro cytotoxic activity against lung cancer cell line A549 using MTT method. IC50 value for all four complexes was determined and all of them are found active. Computational studies of the representative complexes have been done using B3LYP/631-G* basis sets to provide optimized geometry.

Preparation of Self-Assembled, Curcumin-Loaded Nano-Micelles Using Quarternized Chitosan-Vanillin Imine (QCS-Vani Imine) Conjugate and Evaluation of Synergistic Anticancer Effect with Cisplatin.

Nano-micelles are self-assembling colloidal dispersions applied to enhance the anticancer efficacy of chemotherapeutic agents. In this study, the conjugate of quarternized chitosan and vanillin imine (QCS-Vani imine) was synthesized using the reaction of a Schiff base characterized by proton-NMR (1HNMR), UV-Vis spectroscopy, and FT-IR. The critical micelle concentration (CMC), particle size, and zeta potential of the resulting product were determined. The QCS-Vani imine conjugate was used as a carrier for the development of curcumin-loaded nano-micelles, and their entrapment efficiency (%EE), drug-loading capacity (%LC) and in vitro release were investigated using HPLC analysis. Moreover, the nano-micelles containing curcumin were combined with various concentrations of cisplatin and evaluated for a possible anticancer synergistic effect. The anticancer activity was evaluated against lung cancer A549 and mouse fibroblast L929 cell lines. The percent yield (%) of the QCS-Vani imine conjugate was 93.18%. The curcumin-loaded QCS-Vani imine nano-micelles were characterized and found to have a spherical shape (by TEM) with size < 200 nm (by DLS) with high %EE up to 67.61% and %LC up to 6.15 ± 0.41%. The loaded lyophilized powder of the nano-micelles was more stable at 4 °C than at room temperature during 120 days of storage. pH-sensitive release properties were observed to have a higher curcumin release at pH 5.5 (cancer environment) than at pH 7.4 (systemic environment). Curcumin-loaded QCS-Vani imine nano-micelles showed higher cytotoxicity and selectivity toward lung cancer A549 cell lines and exhibited lower toxicity toward the normal cell (H9C2) than pure curcumin. Moreover, the curcumin-loaded QCS-Vani imine nano-micelles exhibited an enhanced property of inducing cell cycle arrest during the S-phase against A549 cells and showed prominently induced apoptosis in lung cancer cells compared to that with curcumin. The co-treatment of cisplatin with curcumin-loaded QCS-Vani imine nano-micelles presented an enhanced anticancer effect, showing 8.66 ± 0.88 μM as the IC50 value, in comparison to the treatment with cisplatin alone (14.22 ± 1.01 μM). These findings suggest that the developed QCS-Vani imine nano-micelle is a potential drug delivery system and could be a promising approach for treating lung cancer in combination with cisplatin.

Synthesis of 1,3-benzodioxole-based procaine-tagged ionic liquids and investigation of its physiochemical and anti-cancer properties against A549 cells

There is significant interest in ionic liquids based on 1,3-benzodioxole as potential leads for novel anticancer agents. Herein, we report the design and development of a series of 1,3-benzodioxole-based procaine-tagged ionic liquids as anticancer drugs in which we investigated the molecular docking studies with tubulin protein followed by synthesis of desired ionic liquids [Pro-pipBF4], [Pro-pipOTF], and [Pro-pipDCN] with binding energy of −355.26 kcal/mol, −335.76 kcal/mol, and −335.39 kcal/mol respectively. Also, in-silico pharmacological and physicochemical properties were assessed for synthesized ionic liquids using admetSAR tool to find out the best candidature for anticancer properties. Further in-vitro studies were performed for their anticancer activity against lung cancer cell line A549 using MTT assay (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide) which revealed the potential effectiveness of [Pro-pipBF4] as an anticancer agent with IC50:19.85 value. As a result, this study evaluated that [Pro-pipBF4] could be a promising therapeutic candidate that may be further developed for cancer treatment.

Blaps rynchopetera combined with cyclophosphamide affects proliferation and apoptosis of lung cancer cells via Wnt/β-catenin signaling pathway

This study aims to investigate the effects of Blaps rynchopetera Fairmaire and/or cyclophosphamide on the proliferation and apoptosis of lung cancer cells and decipher the underlying mechanism. B. rynchopetera and cyclophosphamide-containing serum and blank serum were prepared from SD rats. Cell counting kit-8(CCK-8) assay was employed to examine the proliferation of lung cancer cell lines A549 and Lewis treated with corresponding agents. The Jin's formula method was used to evaluate the combined effect of the two drugs. According to the evaluation results, appropriate drug concentrations and lung cancer cell line were selected for subsequent experiments, which included control, B. rynchopetera, cyclophosphamide, B. rynchopetera + cyclophosphamide, and B. rynchopetera + Wnt/β-catenin pathway agonist lithium chloride(LiCl) groups. Immunocytochemistry was employed to measure the expression of proliferation-related proteins in Lewis cells after drug interventions. Flow cytometry was employed to determine the cell cycle and apoptosis. The expression levels of proliferating cell nuclear antigen(PCNA), cyclinD1, B-cell lymphoma 2(Bcl-2), Bcl-2-assiocated X protein(Bax), Wnt1, and β-catenin were determined by Western blot. The results showed that B. rynchopetera and/or cyclophosphamide significantly inhibited the proliferation of A549 and Lewis cells. Compared with B. rynchopetera alone, the combination increased the inhibition rate on cell proliferation. The combination of B. rynchopetera and cyclophosphamide demonstrated a synergistic effect according to Jin's formula-based evaluation. Compared with the control group, the B. rynchopetera, cyclophosphamide, and B. rynchopetera + cyclophosphamide groups showed increased proportion of Lewis cells in G_0/G_1 phase, increased apoptosis rate, up-regulated expression of Bax, and down-regulated expression of PCNA, cyclinD1, Bcl-2, Wnt1, and β-catenin. Compared with the cyclophosphamide group, the combination group showed increased proportion of cells in G_0/G_1 phase, increased apoptosis rate, up-regulated expression of Bax, and down-regulated expression of PCNA, cyclinD1, Bcl-2, Wnt1, and β-catenin. Compared with the B. rynchopetera group, the B. rynchopetera + LiCl group had deceased proportion of cells in G_0/G_1 phase, decreased apoptosis rate, down-regulated expression of Bax, and up-regulated expression of PCNA, cyclinD1, Bcl-2, Wnt1, and β-catenin. The results indicated that B. rynchopetera could inhibit the proliferation, arrest the cell cycle, and induce the apoptosis of lung cancer cells by inhibiting the Wnt/β-catenin signaling pathway. Moreover, B. rynchopetera had a synergistic effect with cyclophosphamide.

Effects of the PARP Inhibitor Niraparib on the Radiosensitivity of Human Lung and Cervical Cancer Cells

DNA damage repair (DDR) initiated after DNA damage may lead to a decreased radiosensitivity of tumor cells or even radiation resistance. Previous studies indicate that PARP inhibitors block the repair of DNA single-strand breaks (SSBs), and unrepaired SSBs may continuously accumulate and form double-strand breaks (DSBs) that promotes tumor cell death. Studies have also shown that PARP inhibitors act synergistically with RT to inhibit DDR. In this study, we aim to investigate the effect of Niraparib, a clinically approved PARP inhibitor on the radiosensitivity of human lung and cervical cancer cells, and preliminarily explore the underlying mechanism. The human lung cancer cell line A549 and human cervical cancer cell line Siha were both treated with niraparib (for 1h), or X-ray (4Gy), or Niraparib (for 1h) combined with X-ray (4Gy). The effect was examined via measurements of cell proliferation by the cell counting kit-8 assay, and cell viability was detected by clone formation assays. Cell apoptosis and cell cycle distribution were investigated by flow cytometry. In human lung and cervical cancer cell lines, Niraparib combined with radiation therapy significantly inhibited cell proliferation. The proportion of apoptotic cells in cell lines treated with Niraparib plus radiation (the combination group) was significantly higher compared with control, radiation-only, and Niraparib-only groups. Additionally, the proportion of A549 cells in the G2/M phase was significantly increased in the combination group compared with the radiation-only group. The results support that PARP inhibitor Niraparib increases the radiosensitivity of tumor cells, promotes their apoptosis, and induces cell cycle redistribution. The possible mechanism is associated with the inhibition of radiation-induced DNA damage repair.

B-363 Solid Lipid Nanoparticles Mediated CBLC siRNA Delivery as a Platform to Monitor and Probe Biological Functions

Abstract Background Nanoparticles have been used as a tool for delivering therapeutics since few decades. Researchers has been focusing on advances and modifications to improve the properties of these nanoparticles such as sustained drug delivery and bioavailability. This study focuses on modifying the conventional solid lipid nanoparticles (SLN) with Arginine-Glycine-Aspartic acid tripeptide (RGD). The peptidyl link in these SLN has an ability to bind to the integrin of the target cells, allowing solid adhesion to the cells for sustained drug delivery. The use of small interfering RNA (siRNA) allow better invasion through the cell membranes, silencing the intracellular targets. This study uses Casistas B-lymphoma lineage (CBLC) siRNA as the therapeutics for the treatment of non-small cell lung cancer (NSCLC). CBL family consisting of different proteins such as CBL, CBLB, and CBLC origins from a RING finger class. They are E3 ubiquitin ligases responsible for transferring ubiquitin from E2 to E3 substrate. They function as the negative regulator of Tyrosine Kinase activity in cell cycle signaling. CBLC is abundantly present in epithelial cells and so its expression is noteworthy in lung adenocarcinoma. CBL family also possess Tyrosine Kinase Binding domain (TKB) responsible to decrease the activated Epidermal Growth Factor Receptor (aEGFR). Although, CBLC was initially known to perform the same function as other CBL family members, reports suggest that it is a demethylated target. The expression of CBLC upregulates with the treatment of NSCLC by inhibiting agents for DNA methylation. This process increases the demethylation of the CBLC promoter upregulating CBLC. This causes CBLC to compete with CBL for binding to the aEGFR leading to polyubiquitination of aEGFR. Trafficking of aEGFR in the nucleus makes it more stable progressing lung cancer. CBLC silencing allows tumor cells to be sensitized to the therapy with tyrosine kinase inhibiting agents. Methods SLN were modified with RGDP followed by the loading of CBLC siRNA. To characterize the SLN within the diameter range of 20–100 nm, Scanning Electron Microscopy (SEM) was performed. The Energy-dispersive X-ray Spectroscopy was performed to confirm the presence of Phosphorous and Nitrogen in SLN coming from lipids and RNA. Dynamic Light Scattering (DLS) was done to verify the SEM results. To measure the efficacy of the SLN, the cells were treated with control SLN vs siRNA loaded SLN. The Sybr gold assay and invitro release study was done to confirm the release of siRNA from SLN. The human lung normal cell line NL-20 as control and lung cancer cell line A549 were cultured and treated with synthesized nanoparticles for knocking down CBLC gene. The assays such as MTT, western blot, flow cytometry and mass spectrometry-based proteomics were performed to confirm the knockdown of the CBLC gene. Results The SLN characterization confirmed the diameter in the expected range. The efficacy assays confirmed the encapsulation and release of CBLC siRNA. The results from cell studies showed reduction in tumor growth and viability confirming the knockdown of CBLC gene. Conclusion The data suggests significant effects for the use of synthesized nanoparticles as a therapeutic for the treatment of NSCLC.

Metformin induces ferroptosis through the Nrf2/HO-1 signaling in lung cancer

BackgroundMetformin is the most frequently prescribed medication for the treatment of type II diabetes mellitus and has played an anti-tumor potential in a variety of cancer types. Metformin can inhibit the growth of many cancer cells through various mechanisms, including ferroptosis. However, it is still unclear whether metformin can induce ferroptosis in lung cancer.MethodsThis study evaluated the anti-tumor effect of metformin by detecting the levels of oxidative stress factors, the levels of ferrous ions, and the expression of ferroptosis-related genes in A549 and H1299 lung cancer cell lines treated with or without metformin.ResultsThe results showed that metformin treatment increased the levels of MDA, ROS and iron ions, while decreased the levels of GSH, T-SOD and CAT. Meanwhile, metformin treatment reduced the protein expression levels of Gpx4 and SLC7A11, Nrf2 and HO-1, while the addition of ferroptosis inhibitor ferrostatin-1 reversed the reduction.ConclusionsThese results demonstrated that metformin exerts anti-tumor effects by inducing ferroptosis through the Nrf2/HO-1 signaling pathway in lung cancer cells, providing a theoretical basis for drug therapy of lung cancer patients.

Antidiabetic, antioxidant and cytotoxicity activities of ortho- and para-substituted Schiff bases derived from metformin hydrochloride: Validation by molecular docking and in silico ADME studies

Abstract This work evaluates the in vitro antioxidant and antidiabetic activities of two metformin hydrochloride-based Schiff bases. Moreover, the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay was used to examine the in vitro cytotoxic effects of HL1 and HL2 on the A549 lung cancer cell line. The two Schiff bases that have been previously synthesized by using two effective, green techniques, namely stirring and microwave-assisted, are N,N-dimethyl-N′-[(Z)-(2-nitrophenyl) methylidene] imidodicarbonimidic diamide and N,N-dimethyl-N′-[(Z)-(4-nitrophenyl) methylidene] imidodicarbonimidic diamide, indicated by HL1 and HL2, respectively. Studies of antidiabetic efficacy using alpha-amylase revealed that HL2 has a higher inhibition than HL1, but the results on sucrase enzyme showed that HL1 had the highest inhibitory action, whereas the outcome of the antioxidant test with the 2,2-diphenyl-1-picrylhydrazyl assay demonstrated that HL2 was the most effective antioxidant, followed by ascorbic acid and HL1. In the MTT assay, HL1 had the best result, with an IC50 value of 57.13 µg/mL compared to HL2 with an IC50 value of 76.83 µg/mL. It was observed that HL1 was the most effective against the human lung cancer cell line A459. The findings were supported by computational and pharmacokinetic studies (SwissADME). Based on empirical and computational studies, we suggest that HL1 and HL2 are promising candidates as antioxidants and antidiabetics after being examined in vivo.

Luteolin inhibits A549 cells proliferation and migration by down-regulating androgen receptors

BackgroundYi Fei Qing Hua Granules (YQG) is a traditional Chinese herbal medicine with the effects of inhibiting the proliferation of lung cancer cells. Luteolin is one of the active compounds of YQG. Luteolin is a common flavonoid extracted from natural herbs and it can promote cancer cells apoptosis has been reported. However, the underlying molecular mechanism and effects of luteolin on human lung cancer needs to be validated.MethodsMolecular docking, network pharmacology methods and quantitative structure–activity relationship (QSAR) model were used to identify the active components of YQG and their possible mechanisms of action. Western blot analysis was used to measure AR expression in A549 cells. Cell migration assays were used to detect A549 cells proliferation transfected by AR plasmid and AR mutation plasmid, respectively.ResultsTCMSP search results revealed that there are 182 active compounds in YQG, which correspond to 232 target genes. Sixty-one genes were overlapping genes in the 2 datasets of TCMSP and GeneCards. Through bioinformatics tagging of these overlapping genes, a total of 1,951 GO functional tagging analysis and 133 KEGG pathways were obtained. Through molecular docking technology and QSAR model verification, the multi-target active compound luteolin was screened out as one of the active components of YQG for in vitro verification. Androgen receptor (AR) was the hub protein with the highest docking score of luteolin. Western blot showed that luteolin could inhibit AR protein expression in lung cancer cell line A549. After the phosphorylation site of AR protein 877 was inactivated, the ability of luteolin to inhibit the proliferation of lung cancer cells was weakened. Luteolin significantly inhibited the growth of A549 xenogeneic tumors at day 25 and 28 and inhibited the expression of AR.ConclusionIn this study, we have explored luteolin as one of the active components of YQG, and may inhibit the proliferation and migration of A549 cells by decreasing the expression of AR and the regulation of phosphorylation at AR-binding sites.

Nrf2 depletion in the context of loss-of-function Keap1 leads to mitolysosome accumulation

Transcription factor nuclear factor erythroid 2 p45-related factor 2 (Nrf2) is the principal determinant of the cellular redox homeostasis, contributing to mitochondrial function, integrity and bioenergetics. The main negative regulator of Nrf2 is Kelch-like ECH associated protein 1 (Keap1), a substrate adaptor for Cul3/Rbx1 ubiquitin ligase, which continuously targets Nrf2 for ubiquitination and proteasomal degradation. Loss-of-function mutations in Keap1 occur frequently in lung cancer, leading to constitutive Nrf2 activation. We used the human lung cancer cell line A549 and its CRISPR/Cas9-generated homozygous Nrf2-knockout (Nrf2-KO) counterpart to assess the role of Nrf2 on mitochondrial health. To confirm that the observed effects of Nrf2 deficiency are not due to clonal selection or long-term adaptation to the absence of Nrf2, we also depleted Nrf2 by siRNA (siNFE2L2), thus creating populations of Nrf2-knockdown (Nrf2-KD) A549 cells. Nrf2 deficiency decreased mitochondrial respiration, but increased the mitochondrial membrane potential, mass, DNA content, and the number of mitolysosomes. The proportion of ATG7 and ATG3 within their respective LC3B conjugates was increased in Nrf2-deficient cells with mutant Keap1, whereas the formation of new autophagosomes was not affected. Thus, in lung cancer cells with loss-of-function Keap1, Nrf2 facilitates mitolysosome degradation thereby ensuring timely clearance of damaged mitochondria.

Hydroxychloroquine-Loaded Chitosan Nanoparticles Induce Anticancer Activity in A549 Lung Cancer Cells: Design, BSA Binding, Molecular Docking, Mechanistic, and Biological Evaluation.

The current study describes the encapsulation of hydroxychloroquine, widely used in traditional medicine due to its diverse pharmacological and medicinal uses, in chitosan nanoparticles (CNPs). This work aims to combine the HCQ drug with CS NPs to generate a novel nanocomposite with improved characteristics and bioavailability. HCQ@CS NPs are roughly shaped like roadways and have a smooth surface with an average size of 159.3 ± 7.1 nm, a PDI of 0.224 ± 0.101, and a zeta potential of +46.6 ± 0.8 mV. To aid in the development of pharmaceutical systems for use in cancer therapy, the binding mechanism and affinity of the interaction between HCQ and HCQ@CS NPs and BSA were examined using stopped-flow and other spectroscopic approaches, supplemented by molecular docking analysis. HCQ and HCQ@CS NPs binding with BSA is driven by a ground-state complex formation that may be accompanied by a non-radiative energy transfer process, and binding constants indicate that HCQ@CS NPs-BSA was more stable than HCQ-BSA. The stopped-flow analysis demonstrated that, in addition to increasing BSA affinity, the nanoformulation HCQ@CS NPS changes the binding process and may open new routes for interaction. Docking experiments verified the development of the HCQ-BSA complex, with HCQ binding to site I on the BSA structure, primarily with the amino acids, Thr 578, Gln 579, Gln 525, Tyr 400, and Asn 404. Furthermore, the nanoformulation HCQ@CS NPS not only increased cytotoxicity against the A549 lung cancer cell line (IC50 = 28.57 ± 1.72 μg/mL) compared to HCQ (102.21 ± 0.67 μg/mL), but also exhibited higher antibacterial activity against both Gram-positive and Gram-negative bacteria when compared to HCQ and chloramphenicol, which is in agreement with the binding constants. The nanoformulation developed in this study may offer a viable therapy option for A549 lung cancer.

Flexibanone, the first cembranoid possessing an α-methylene-δ-lactone linking with a tetrahydronfuran ring by C-3/C-4 single bond, from the soft coral Sinularia flexibilis

A new cembranoid, flexibanone (1), and a known related diterpene sinulariolone (2) were isolated from a Taiwanese soft coral Sinularia flexibilis. Compound 1 is the first cembranoid possessing an α-methylene-δ-lactone linking with a tetrahydronfuran ring by C-3/C-4 single bond. The structures of 1 and 2 were established by using extensive spectroscopic analysis, including 2D NMR methods. Both compounds were suggested to be biosynthesized from the same cembranoid precursor. Further, the cytotoxicity against human oral cancer Ca9-22, breast adenocarcinoma MDA-MB-231 and non-small cell lung cancer A549 cell lines of 1 and 2 were also assayed.

Half-Sandwich Iridium(III), Rhodium(III), and Ruthenium(II) Complexes Chelating Hybrid sp2-N/sp3-N Donor Ligands to Achieve Improved Anticancer Selectivity.

The biological efficacy of half-sandwich platinum group organometallic complexes of the formula [(η5-Cpx)/(η6-arene)M(XY)Cl]0/+ (XY = bidentate ligands; Cpx = functionalized cyclopentadienyl; M = Ir, Rh, Ru, Os) has received considerable attention due to the significance of the metal center, chelating ligand, and Cpx/arene moieties in defining their anticancer potency and selectivity. With a facile access to the BIAN-derived imine-amine ligands using alkylaluminum as the reductant, we herein described the preparation and characterization of 16 half-sandwich Ir(III), Rh(III), and Ru(II) complexes chelating the hybrid sp2-N/sp3-N donor ligand. A nonplanar five-member metallacycle was confirmed by X-ray single-crystal structures of Ir1-Ir3, Ir7, Rh1, Ru1, and Ru4. The attempt to prepare imine-amido complexes using a base as the deprotonating agent led to the mixture of imine-amine complexes, within which the leaving group Cl- was displaced, and 16-electron imine-amido complexes without Cl-. The half-sandwich imine-amine complexes in this system underwent rapid hydrolysis in aqueous solution, exhibited weak photoluminescence, and showed the ability of binding to CT-DNA and BSA. The cytotoxicity of all imine-amine complexes against A549 lung cancer cell lines, HeLa cervical cancer cell lines, and 4T1 mouse breast cancer cells was determined by an MTT assay. The IC50 values of these complexes were in a range of 5.71-67.28 μM. Notably, most of these complexes displayed improved selectivity toward A549 cancer cells versus noncancerous BEAS-2B cells in comparison with the corresponding α-diimine complexes chelating the sp2-N/sp2-N donor ligand, which have been shown no selectivity in our previous report. The anticancer selectivity of these complexes appeared to be related to the redox-based mechanism including the catalytic oxidation of NADH to NAD+, reactive oxygen species (ROS) generation, and depolarization of the mitochondrial membrane. Further, inducing apoptosis of these complexes in A549 cancer cells and BEAS-2B normal cells also correlated with their anticancer selectivity, indicating the apoptosis mode of cell death in this system. In addition, these complexes could enter A549 cells via energy-dependent pathway and were able to impede the in vitro migration of A549 cells.

IN VITRO ANTICANCER POTENTIAL OF PHYTOGENIC AG-AU BIMETALLIC NANOPARTICLES USING CLITORIA TERNATEA FLOWER EXTRACT

Bimetallic nanoparticles (BNPs) have gained significant attention in the field of biomedical and pharmaceutical because of its tunable size, shape, high surface to volume and enhanced biological properties. In this study, we report the ecofriendly route to produce the noble (Ag-Au) bimetallic nanoparticles using aqueous flower extract of Clitoria ternatea as a reducing and capping agent. The synthesized C.T-Ag-Au BNPs were characterized by using physicochemical techniques such as UV-visible, FTIR, XRD and SEM-EDX analysis. The UV-visible spectra reveal the formation of homogeneous bimetallic nanoparticles by the single blue-shifted peak at 541 nm of C.T-Ag-Au BNPs. The phyto fabrication of synthesized C.T-Ag-Au BNPs was analyzed using FTIR spectroscopy. XRD confirms the formation of phase pure cubic Ag-Au alloy bimetallic nanoparticles with crystallite size is 14.5 nm. The surface morphology and elemental analysis of C.T-Ag-Au BNPs were examined by using SEM-EDX analysis. The synthesized C.TAg- Au BNPs were evaluated for their cytotoxicity against A549 human lung cancer cells through standard MTT assay. From this assay, green synthesized bimetallic nanoparticles induced cell apoptosis, suggesting that the synthesized C.T-Ag-Au BNPs gave best anticancer properties against lung cancer A549 cell lines.

Anlotinib affects systemic lipid metabolism and induces lipid accumulation in human lung cancer cells

BackgroundAnlotinib has demonstrated encouraging clinical outcomes in the treatment of lung cancer, soft tissue sarcoma and thyroid carcinoma. Several clinical studies have shown a relationship between anlotinib treatment and the occurrence of hyperlipidemia. The fundamental mechanisms, however, are still largely unclear. Here, the effect of anlotinib on lipid metabolism in an animal model and human cancer cells was evaluated and the role of lipid metabolism in the antitumor efficacy of anlotinib was investigated.MethodsThe C57BL/6 J mouse model as well as A549 and H460 human lung cancer cell lines were used to examine the impact of anlotinib on lipid metabolism both in vivo and in vitro. Levels of triglycerides, high-density lipoprotein, low-density lipoprotein (LDL), and total cholesterol in serum or cell samples were determined using assay kits. The expression levels of crucial genes and proteins involved in lipid metabolism were measured by quantitative RT-PCR and Western blotting. Furthermore, exogenous LDL and knockdown of low-density lipoprotein receptor (LDLR) were used in H460 cells to investigate the relevance of lipid metabolism in the anticancer efficacy of anlotinib.ResultsAnlotinib caused hyperlipidemia in C57BL/6 J mice, possibly by downregulating hepatic LDLR-mediated uptake of LDL cholesterol. AMP-activated protein kinase and mammalian target of rapamycin inhibition may also be involved. Additionally, anlotinib enhanced sterol response element binding protein 1/2 nuclear accumulation as well as upregulated LDLR expression in A549 and H460 cells, which may be attributable to intracellular lipid accumulation. Knockdown of LDLR reduced intracellular cholesterol content, but interestingly, anlotinib significantly improved intracellular cholesterol accumulation in LDLR-knockdown cells. Both exogenous LDL and LDLR knockdown decreased the sensitivity of cells to anlotinib.ConclusionsAnlotinib modulates host lipid metabolism through multiple pathways. Anlotinib also exerts a significant impact on lipid metabolism in cancer cells by regulating key transcription factors and metabolic enzymes. In addition, these findings suggest lipid metabolism is implicated in anlotinib sensitivity.

A novel quinoline derivative, DFIQ, sensitizes NSCLC cells to ferroptosis by promoting oxidative stress accompanied by autophagic dysfunction and mitochondrial damage

BackgroundThe development of nonapoptotic programmed cell death inducers as anticancer agents has emerged as a cancer therapy field. Ferroptosis, ferrous ion-driven programmed cell death that is induced by redox imbalance and dysfunctional reactive oxygen species (ROS) clearance, is triggered during sorafenib and PD-1/PD-L1 immunotherapy. DFIQ, a quinoline derivative, promotes apoptosis by disrupting autophagic flux and promoting ROS accumulation. Our pilot experiments suggest that DFIQ participates in ferroptosis sensitization. Thus, in this study, we aimed to reveal the mechanisms of DFIQ in ferroptosis sensitization and evaluate the clinical potential of DFIQ.MethodsWe treated the non-small cell lung cancer (NSCLC) cell lines H1299, A549, and H460 with the ferroptosis inducer (FI) DFIQ and analyzed viability, protein expression, ROS generation, and fluorescence staining at different time points. Colocalization analysis was performed with ImageJ.ResultsDFIQ sensitized cells to FIs such as erastin and RSL3, resulting in a decrease in IC50 of at least 0.5-fold. Measurement of ROS accumulation to explore the underlying mechanism indicated that DFIQ and FIs treatment promoted ROS accumulation and SOD1/SOD2 switching. Mitochondria, known ROS sources, produced high ROS levels during DFIQ/FI treatment. RSL3 treatment promoted mitochondrial damage and mitophagy, an autophagy-associated mitochondrial recycling system, and cotreatment with DFIQ induced accumulation of mitochondrial proteins, which indicated disruption of mitophagic flux. Thus, autophagic flux was measured in cells cotreated with DFIQ. DFIQ treatment was found to disrupt autophagic flux, leading to accumulation of damaged mitochondria and eventually inducing ferroptosis. Furthermore, the influence of DFIQ on the effects of clinical FIs, such as sorafenib, was evaluated, and DFIQ was discovered to sensitize NSCLC cells to sorafenib and promote ferroptosis.ConclusionsThis study indicates that DFIQ not only promotes NSCLC apoptosis but also sensitizes cells to ferroptosis by disrupting autophagic flux, leading to accumulation of dysfunctional mitochondria and thus to ferroptosis. Ferroptosis is a novel therapeutic target in cancer therapy. DFIQ shows the potential to enhance the effects of FIs in NSCLC and act as a potential therapeutic adjuvant in ferroptosis-mediated therapy.

Triterpenoids from the Stem Bark of &lt;i&gt;Aglaia cucullata&lt;/i&gt; (Meliaceae) and Their Cytotoxic Activity against A549 Lung Cancer Cell Line

The Aglaia species, which contains triterpenoids, is the most numerous in the Meliaceae family. The A. cucullata species, of which there are only a few known examples, has received scant research attention. This investigation aims to identify triterpenoids in an n-hexane preparation of A. cucullata stem bark and evaluate their effects against the A549 lung cancer cell line. Five dammarane-type triterpenoids were isolated from the A. cucullata trunk bark, which is (1) (20S)-20-hydroxydammar-24-en-3-one, (2) cabraleone, (3) cabralealactone, (4) eichlerianic acid, and (5) (+)-fouquierol. Their chemical structures were determined using infrared, high-resolution mass spectrometry, and nuclear magnetic resonance, as well as through data comparison of the reported compounds. Compound 1 was priorly separated from the Aglaia genus, compounds 2–4 were first isolated from the A. cucullata species, and compound 5 has been reportedly isolated from the Meliaceae family and the Aglaia genus. All substances were tested for their lethal potential against the A549 lung cancer cell type. A seco structure in the A ring of dammarane-type triterpenoid might play an important part in the lethal activity of component 4, which showed the greatest activity with an IC50 value of 32.17 µM against the A549 lung cancer cell line.

In vitro anticancer screening of herbal syrup formulation of turmeric extract

In continuation of previous research, the anticancer screening of formulation of Herbal syrup of Turmeric extract was performed. The formulation of herbal syrup was performed as per reported literature method. The anticancer cancer screening was performed on two Human cancer cell lines. These cell lines were Human Lung Cancer cell line A549 and Human Breast Cancer Cell line MCF7. The formulation was found to be devoid of anticancer properties.

In vitro anticancer screening of herbal syrup formulation of vinca extract

In continuation of previous research, the anticancer screening of formulation of herbal syrup of Vinca extract was performed. The formulation of herbal syrup was performed as per reported literature method. The anticancer cancer screening was performed on two Human cancer cell lines. These cell lines were Human Lung Cancer cell line A549 and Human Breast Cancer Cell line MCF7. The formulation was found to be devoid of anticancer properties.