Human Breast Adenocarcinoma Cell Line Research Articles

Synthesis, Characterization, and Biological Investigation of Antimony(III) Halide Complexes With Different Coordination Architectures Constructed From Thiophene Thiosemicarbazones

ABSTRACTIn this study, we report the synthesis, characterization, and biological investigation of antimony(III) halide complexes with various coordination architectures constructed from thiophene thiosemicarbazones. Antimony(III) thiophene‐2‐carbaldehyde thiosemicarbazone complexes (1, 2, 4, and 5) exhibit a square pyramidal geometry, with ligands coordinated to the central antimony atom in two distinct binding modes. In contrast, antimony(III) 2‐acetylthiophene thiosemicarbazone complexes (3, 6, and 7) adopt a seesaw geometry. These complexes (1–7) represent the first reported examples of antimony(III) halide thiosemicarbazone compounds. The unique coordination environments observed in these complexes are of significant importance within the realm of antimony chemistry. These synthesized complexes exhibit different coordination geometries as well as potential biological activities. The antiproliferative activity against the human breast adenocarcinoma (MCF‐7) cell line and antimicrobial activity against Gram‐positive and Gram‐negative bacteria were assessed. Antimony(III) thiophene‐2‐carbaldehyde thiosemicarbazone complexes (1, 2, 4, and 5) showed significant antiproliferative activity with IC50 values ranging from 8.5 to 19.1 μM, while antimony(III) 2‐acetylthiophene thiosemicarbazone complexes (3, 6, and 7) had higher IC50 values. Additionally, the antimony complexes demonstrated selective antimicrobial activity against Gram‐negative bacteria.

Boron Salicylate Ester Compounds as Boron Therapeutics. Their Synthesis, Structural Characterizations and Anticancer Effects against MDA-MB-231.

The element boron forms a wide range of borate minerals with different properties. Borate minerals make it possible to design boron-containing molecules with new biological properties in terms of their chemical structure and properties. It is known that boron compounds have antioxidant, anti-inflammatory, anti-tumor and anti-cancer properties. This makes boron compounds important for the future development of boron chemotherapeutics, boron supplements and new drugs. Reliable scientific studies on boron compounds will facilitate the clear presentation of their functions in its biological applications and metabolism. In this study, boron monoester and boron diester structures were synthesized with salicylic acid ligand. To stabilize boron ester structures, Na+, K+, Mg2+, Ca2+ cations were used as counter-ions. Structural properties of the synthesized substances, molecules obtained by crystallization/precipitation from aqueous solutions in solid state, elemental analysis, melting point determination, infrared spectroscopy analysis (FT-IR), thermal analysis (TGA/DTA), mass analysis (GC-MS) and single crystal analysis. Structural properties were tried to be explained by structure analysis (SC-XRD) methods. Additionally, the anticancer potential of boron salicylate esters against the MDA-MB-231 human breast adenocarcinoma cell line was examined. The K-B salicylate diester molecule was found to have the most potential potency with the lowest IC50 value against the MDA-MB-231 cell line. The anticancer potential of boron salicylate esters can be further investigated with other cancer models with the combination of anticancer drugs. It is also thought that the mechanism of action of these molecules may help reveal their further applications.

Copper(II), zinc(II) and cadmium(II) complexes with the O-picolyl ether of chiral α-aminooxime of natural (+)-limonene: synthesis, structures, luminescence and cytotoxicity

Copper(II), zinc(II) and cadmium(II) complexes, [CuLCl2], [ZnLCl2] and [CdLCl2], with a new derivative of chiral α-aminooxime of natural (+)-limonene bearing a pycolyl group (L) were synthesized. The complexes are mononuclear, the L molecules act as N,N,N-tridentate chelating ligands. The computations support the fact that the N,N,N–chelation by a metal ion is thermodynamically preferred over the N,N–chelation for all complexes. The ligand L and the complexes [ZnLCl2] and [CdLCl2] exhibit fluorescence, with the emission spectra depending on the excitation wavelength. L emits in close to white region, while the Zn(II) and Cd(II) complexes demonstrate blue emission. Due to chelation-induced fluorescence enhancement effect, the fluorescence quantum yield increases when going from L to [ZnLCl2] and [CdLCl2]. The complex [CdLCl2] displays cytotoxicity against the HepG2 cell line (IC50 = 14.3 ± 1.5 μM), which is almost twice as high as that of cisplatin. The human breast adenocarcinoma cell line (MCF-7) is more sensitive to [CuLCl2] (IC50 = 43.4 ± 0.4 μM) than to [CdLCl2].

The antioxidant and selective apoptotic activities of modified auraptene-loaded graphene quantum dot nanoparticles (M-AGQD-NP)

BackgroundPancreatic and Gastric cancers are very aggressive and deadly types of cancer that require effective treatment strategies to stop their progression. Nano-drug delivery systems, like those using Auraptene-loaded GQD nanoparticles, play a crucial role in addressing this need by delivering targeted and controlled treatments to cancer cells, making treatment more effective, and reducing side effects. The study focused on investigating the effects of Auraptene, an efficient anticancer compound when loaded into Graphene Quantum Dots (GQDs) on types of human cancer cells.MethodsTo create auraptene-loaded graphene quantum dot nanoparticles (AGQD-NP) (Unmodified and modified types) a combination of hydrothermal and high-energy homogenization methods was used. The nanoparticles were characterized by conducting DLS (Dynamic light scattering), FTIR (Fourier-transform infrared spectroscopy), FESEM (Field Emission Scanning Electron microscopy), and zeta potential analysis. bioactivity of AGQD-NP was assessed through tests, including antioxidant capacity measured by ABTS and DPPH scavenging abilities well as cytotoxicity tested using MTT assay on both human cancer cell lines and normal human vascular endothelial cells.ResultsThe modified AGQD-NP (M-AGQD-NP) demonstrated antioxidant properties by neutralizing free radicals. They also displayed selective toxicity, towards human gastric adenocarcinoma cell-line (AGS) and human pancreatic adenocarcinoma (PANC) cancer cells with IC50 values recorded at 78.8 µg/mL and 89.72 µg/mL respectively. The specific targeting of gastric cancer cells was evident from the differing IC50 values compared to the Human breast adenocarcinoma cell line (MCF-7), Human hepatocellular carcinoma cell line (Hella), and normal vascular endothelial cells (Huvec). Additionally, the induced apoptotic death, in the human pancreatic adenocarcinoma (PANC) cancer cells was confirmed through AO/PI staining and Annexin-based flow cytometry revealing increased expression levels of P53, Caspase3, BAX, and Caspase8.ConclusionIn summary, the M-AGQD-NP have shown encouraging effects displaying antioxidant capabilities and a specific focus, on pancreatic and gastric cancer cells. These findings indicate uses for AGQD-NP as an efficient apoptosis inducer in cancer treatment. Additional In-vivo researches are required to validate their effectiveness, in living organisms.

Antiproliferative and Pro-apoptotic Activities of Tournefortia mutabilis vent. Leaves on the Human Breast Adenocarcinoma Cell Line (MCF-7)

Background: Breast cancer is the most common cancer among women worldwide, impacting not only the patients but also their families and communities. Tournefortia mutabilis vent. is a plant endemic to Mexico, traditionally used in Zapotec medicine for the treatment of cancer. Objectives: This study aims to evaluate the effects of the chloroformic extract of T. mutabilis vent. leaves on cell proliferation and cell death in MCF-7 cells. Methods: The effect of the extract on MCF-7 cell proliferation was measured using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and crystal violet staining. Apoptosis was evaluated through fluorescein diacetate/propidium iodide staining and caspase-3, -6, and -9 activity assays. Results: The half-maximal inhibitory concentration (IC50) of the T. mutabilis vent. extract on MCF-7 cell proliferation at 48 hours and 72 hours after treatment was 86.4 µg/mL and 2.74 µg/mL, respectively. We observed that the extract and its semi-purified fractions induced cell death through the activation of caspases 3, -6, and -9. Conclusions: Tournefortia mutabilis vent. is a potential source of compounds with antiproliferative and pro-apoptotic activities on the MCF-7 cell line, primarily through the intrinsic pathways of apoptosis.

Preparation, Characterization, DFT Calculations, biological evaluations and Molecular docking studies of benzimidazolium-based N-heterocyclic carbenes and selenium-adducts

This study involves designing through DFT, synthesis, and characterization of three benzimidazolium salts (AL1-AL3) and their corresponding selenium adducts (AC1-AC3). The compounds were characterized theoretically as well as experimentally through FT-IR and 1H &13C NMR. Theoretical calculations were performed using the B3LYP/6–31 G (d, p) levels of theory to determine various optical and chemical properties of the designed molecules. The HOMO/LUMO energies and their energy gaps were also calculated to assess the structure–activity relationships. In vitro testing against the Human breast Adenocarcinoma cell line (MCF-7) was conducted using the MTT assay and 5-Fluorouracil (standard) was used for the comparison of results. The ligand AL3 showed maximum inhibitory potential (66.51 ± 0.82) even approaching the standard 5FU while the complex AC3 showed maximum viability with the least inhibition value as (21.7 ± 0.73) against MCF7 cell line. The antioxidant potential for scavenging DPPH radicals was also assessed, revealing that AL1 has the greatest potential, with a value of 62.01 ± 0.9. The interactions of the complexes with various proteins were also assessed via molecular docking studies, revealing strong binding energies and ligand affinities towards angiogenic factors such as VEGF-A, EGF, HIF, and COX-1. This suggests that the anticancer activities of the complexes AC1-AC3 may be attributed to their potent anti-angiogenic effects. Moreover, selenium-NHC adducts exhibited more significant anticancer potential as compared to their ligands. Furthermore, the antibacterial and antifungal potential of compounds AC1-AC3 was assessed. Notably, the ligand AL2 exhibited significant antibacterial activity against E. coli, as evidenced by a zone of inhibition (ZI) value of 17.5 ± 0.29 mm, whereas AL3 demonstrated the highest activity against P. multocida showing the ZOI value of 18.5 ± 0.38. Among all the synthesized compounds, AL1 displayed the most significant antifungal activity against F. avenaceum (27 ± 0.51 mm), surpassing even the standard drug gentamycin (13 ± 0.23 mm), which was tested under the same conditions. However, its corresponding Se-NHC, AC1, adduct was found to be inactive. Overall, AL3 and its corresponding Se-NHC adduct (AC3), demonstrated superior biological potential against the majority of the tested strains.

Therapeutic Senolysis of Axitinib-Induced Senescent Human Lung Cancer Cells.

Tyrosine kinase inhibitors (TKIs) inhibit receptor-mediated signals in cells. Axitinib is a TKI with high specificity for vascular endothelial growth factor receptors (VEGFRs). We determined whether axitinib could induce senescence in human cancer cells and be lysed by the senolytic drug ABT-263. Human lung and breast adenocarcinoma cell lines were used. These cells were cultured with axitinib or a multi-target TKI lenvatinib. The expression of β-galactosidase, VEGFRs, Ki-67, reactive oxygen species (ROS) of cancer cells, and their BrdU uptake were evaluated by flow cytometry. The mRNA expression of p21 and IL-8 was examined by quantitative PCR. The effects of TKIs on phosphorylation of Akt and Erk1/2, as downstream molecules of VEGFR signaling, were examined by immunoblot. The in vivo anti-cancer effect was examined using a xenograft mice model. Axitinib, but not lenvatinib, induced cellular senescence (increased cell size and enhanced expression of β-galactosidase) in all adenocarcinoma cell lines. Axitinib-induced senescence was unrelated to the expression of VEGFRs on cancer cells. ROS were involved in axitinib-induced senescence. Axitinib-induced senescent lung adenocarcinoma A549 cells were drastically lysed by ABT-263. In A549-xenografted mice, combination therapy with axitinib and ABT-263 significantly suppressed tumor growth with the induction of apoptotic cancer cells.

Combining In Vitro, In Vivo, and Network Pharmacology Assays to Identify Targets and Molecular Mechanisms of Spirulina-Derived Biomolecules against Breast Cancer.

The current research employed an animal model of 7,12-dimethylbenz(a)anthracene (DMBA)-induced mammary gland carcinogenesis. The estrogen receptor-positive human breast adenocarcinoma cell line (MCF-7) was used for in vitro analysis. This was combined with a network pharmacology-based approach to assess the anticancer properties of Spirulina (SP) extract and understand its molecular mechanisms. The results showed that the administration of 1 g/kg of SP increased the antioxidant activity by raising levels of catalase (CAT) and superoxide dismutase (SOD), while decreasing the levels of malonaldehyde (MDA) and protein carbonyl. A histological examination revealed reduced tumor occurrence, decreased estrogen receptor expression, suppressed cell proliferation, and promoted apoptosis in SP protected animals. In addition, SP disrupted the G2/M phase of the MCF-7 cell cycle, inducing apoptosis and reactive oxygen species (ROS) accumulation. It also enhanced intrinsic apoptosis in MCF-7 cells by upregulating cytochrome c, Bax, caspase-8, caspase-9, and caspase-7 proteins, while downregulating Bcl-2 production. The main compounds identified in the LC-MS/MS study of SP were 7-hydroxycoumarin derivatives of cinnamic acid, hinokinin, valeric acid, and α-linolenic acid. These substances specifically targeted three important proteins: ERK1/2 MAPK, PI3K-protein kinase B (AKT), and the epidermal growth factor receptor (EGFR). Network analysis and molecular docking indicated a significant binding affinity between SP and these proteins. This was verified by Western blot analysis that revealed decreased protein levels of p-EGFR, p-ERK1/2, and p-AKT following SP administration. SP was finally reported to suppress MCF-7 cell growth and induce apoptosis by modulating the PI3K/AKT/EGFR and MAPK signaling pathways suggesting EGFR as a potential target of SP in breast cancer (BC) treatment.

Novel tetraaza macrocyclic Schiff base complexes of bivalent zinc: microwave-assisted green synthesis, spectroscopic characterization, density functional theory calculations, molecular docking studies, in vitro antimicrobial and anticancer activities.

In the present manuscript, novel macrocyclic Schiff base complexes [Zn(N4MacL1)Cl2-Zn(N4MacL3)Cl2] were synthesized by the reaction of ZnCl2 and macrocyclic ligands (N4MacL1-N4MacL3) derived from diketone and diamines under microwave irradiation method and conventional method. The structures of the obtained complexes were identified by various spectrometric methods such as Fourier transformation infra-red (FT-IR), nuclear magnetic resonance (NMR), high-resolution mass spectrometry (HR-MS), powder X-ray diffraction, molar conductivity, and UV-vis. The structures of the synthesized compounds were optimized by using the def2-TZV/J and def2-SVP/J Coulomb fitting basis sets at B3LYP level in density functional theory (DFT) calculations. The macrocyclic Schiff base complexes exhibited higher activities against Gram-positive bacteria (Staphylococcus aureus and Bacillus cereus), Gram-negative bacteria (Escherichia coli and Xanthomonas campestris), and fungal strains (Fusarium oxysporum and Candida albicans) in comparison to macrocyclic Schiff base ligands. Furthermore, the newly synthesized macrocyclic compounds were assessed for their anticancer activity against three cell lines: A549 (human alveolar adenocarcinoma epithelial cell line), HT-29 (human colorectal adenocarcinoma cell line), and MCF-7 (human breast adenocarcinoma cell line) using the MTT assay. The obtained results showed that the macrocyclic complex [Zn(N4MacL3)Cl2] displayed the highest cytotoxic activity (2.23 ± 0.25µM, 6.53 ± 0.28µM, and 7.40 ± 0.45µM for A549, HT-29, and MCF-7 cancer cell lines, respectively). Additionally, molecular docking investigations were conducted to elucidate potential molecular interactions between the synthesized macrocyclic compounds and target proteins. The results revealed a consistent agreement between the docking calculations and the experimental data.

Design of Polymeric Nanoparticles for Theranostic Delivery of Capsaicin as Anti-Cancer Drug and Fluorescent Nitrogen-Doped Graphene Quantum Dots.

In recent years, multifunctional nanocarriers that provide simultaneous drug delivery and imaging have attracted enormous attention, especially in cancer treatment. In this research, we designed a biocompatible fluorescent multifunctional nanocarrier for the co-delivery of capsaicin (CPS) and nitrogen-doped graphene quantum dots (N-GQDs) using the pH sensitive amphiphilic block copolymer (poly(2-ethyl-2-oxazoline)-b-poly(ε-caprolactone), PEtOx-b-PCL). The effects of the critical formulation parameters (the amount of copolymer, the concentration of poly(vinyl alcohol) (PVA) as a stabilizing agent in the inner aqueous phase, and volume of the inner phase) were evaluated to achieve optimal nanoparticle properties using Central Composite Design (CCD). The optimized nanoparticles demonstrated a desirable size distribution (167.8±1.4nm) with a negative surface charge (-19.9±0.4) and a suitable loading capacity for capsaicin (70.80±0.05%). The CPS & N-GQD NPs were found to have remarkable toxicity on human breast adenocarcinoma cell line (MCF-7). The solid fluorescent signal was acquired from cells containing multifunctional nanoparticles, according to the confocal microscope imaging results, confirming the significant cellular uptake. This research illustrates the enormous potential for cellular imaging and enhanced cancer therapy offered by multifunctional nanocarriers that combine drug substances with the novel fluorescent agents. This article is protected by copyright. All rights reserved.

Ephrin type-A receptor 2-antisense RNA1/2 promote proliferation and migration of MDA-MB-231 cells through EPHA2-dependent Ras signaling pathway mediated by MAPK8/JNK1, MAPK9/JNK2-NFATC2/NFAT1 and JUND

Ephrin type-A receptor 2 (EPHA2) is a receptor tyrosine kinase that is overexpressed in a variety of cancers, including breast cancer. EPHA2 expression may be causally related to tumorigenesis; therefore, it is important to understand how EPHA2 expression is regulated. We previously reported that EPHA2 antisense RNA (EPHA2-AS), a natural antisense transcript, is an important modulator of EPHA2 mRNA levels and hence production of EPHA2 protein. EPHA2-AS encodes two splice variants, EPHA2-AS1 and EPHA2-AS2. The two variants are constitutively expressed in a concordant manner with EPHA2 mRNA in human breast adenocarcinoma cell lines and in patient samples, with the highest levels detected in the basal-like/triple-negative molecular subtype of breast cancer cells. In this study, we investigated the mechanism of EPHA2-AS1/2 in triple-negative breast cancer using MDA-MB-231 cells. We performed RNA-seq transcriptome analyses of MDA-MB-231 cells treated with AHCC®, which suppressed expression of EPHA2-AS1/2 and EPHA2 mRNA, and EPHA2-AS1/2-silenced MDA-MB-231 cells. Bioinformatics analyses identified 545 overlapping differentially expressed genes that were significantly up- or down-regulated by these treatments. Subsequent functional enrichment analyses of the overlapping genes in combination with in vitro assays indicated that EPHA2-AS1/2 may promote the proliferation and migration of MDA-MB-231 cells through the EPHA2-dependent Ras signaling pathways mediated by MAPK8/JNK1, MAPK9/JNK2-NFATC2/NFAT1 (proliferation and migration) and JUND (migration). These results thus suggest that EPHA2-AS1/2 may represent a potential molecular target for triple-negative breast cancer treatment.

Bcl-2 dependent modulation of Hippo pathway in cancer cells

IntroductionBcl-2 and Bcl-xL are the most studied anti-apoptotic members of Bcl-2 family proteins. We previously characterized both of them, not only for their role in regulating apoptosis and resistance to therapy in cancer cells, but also for their non-canonical functions, mainly including promotion of cancer progression, metastatization, angiogenesis, and involvement in the crosstalk among cancer cells and components of the tumor microenvironment. Our goal was to identify transcriptional signature and novel cellular pathways specifically modulated by Bcl-2.MethodsWe performed RNAseq analysis of siRNA-mediated transient knockdown of Bcl-2 or Bcl-xL in human melanoma cells and gene ontology analysis to identify a specific Bcl-2 transcriptional signature. Expression of genes modulated by Bcl-2 and associated to Hippo pathway were validated in human melanoma, breast adenocarcinoma and non-small cell lung cancer cell lines by qRT-PCR. Western blotting analysis were performed to analyse protein expression of upstream regulators of YAP and in relation to different level of Bcl-2 protein. The effects of YAP silencing in Bcl-2 overexpressing cancer cells were evaluated in migration and cell viability assays in relation to different stiffness conditions. In vitro wound healing assays and co-cultures were used to evaluate cancer-specific Bcl-2 ability to activate fibroblasts.ResultsWe demonstrated the Bcl-2-dependent modulation of Hippo Pathway in cancer cell lines from different tumor types by acting on upstream YAP regulators. YAP inhibition abolished the ability of Bcl-2 to increase tumor cell migration and proliferation on high stiffness condition of culture, to stimulate in vitro fibroblasts migration and to induce fibroblasts activation.ConclusionsWe discovered that Bcl-2 regulates the Hippo pathway in different tumor types, promoting cell migration, adaptation to higher stiffness culture condition and fibroblast activation. Our data indicate that Bcl-2 inhibitors should be further investigated to counteract cancer-promoting mechanisms.

Anabasis articulata (Forssk.) Moq. food aqueous extract triggers oxidative stress-induced senescence and reduces metastatic power in MDA-MB-231 cells

Ancient ethnobotanical practices handed down through traditional knowledge are still commonly employed to treat various pathologies, although the scientific reasons underlying their biological effects have not been clarified yet. In this contribution, the potential antitumoral activity of the aqueous extract from A. articulata (AAE) was investigated to validate the hypothesis of the Algerian folk medicine which would suggest this plant derivative as a functional food for treating breast cancer. A. articulata phytocomplex, isolated by maceration following exactly the African recipe, has been already characterized by our research group in previous works. Thus, the antiproliferative function of AAE against MDA-MB-231, a highly aggressive human breast adenocarcinoma cell line, was evaluated. Slowing down of cell growth, absence of cytotoxicity and DNA fragmentation, and cell cycle arrest at the G2/M phase were observed after treatment with AAE at different doses (0.3–6 mg of dried plant material equivalent per mL of culture medium) for 24 and 48 h. Wound and transwell assays proved that AAE possessed both antimigration and antiinvasive capacities, evidence also supported by molecular analyses focused on Metalloproteases (MMP-2 and MMP-9), Vimentin and ανβ3-Integrin. These results, together with the demonstration of the activation of p53/p21WAF1/Cip1/p27Kip1 pathway and the increase of oxygen reactive species levels, suggested that AAE triggered a senescence process. The final confirmation was obtained by a specific kit staining senescent cells. All our data would explain the efficacy of the Algerian medicinal remedy based on the intake of the investigated functional plant food and would highlight the basics for developing novel natural pharmacological products based on AAE and showing preventive and therapeutic antineoplastic potentialities against highly aggressive breast cancers.

Polycaprolactone Microsphere‐Encapsulated Fluconazole‐Loaded Zinc Oxide and Hydroxyapatite Nanocomposites for Enhanced Biological Performance

AbstractOsteomyelitis (OM), a debilitating disease caused by a microbial infection of the bones, continues to pose a formidable obstacle for orthopedic surgeons. The conventional methods for the prevention and treatment of OM are insufficient. In this research work, we developed a strategic green nanostructure using the antifungal drug Fluconazole (FCZ)‐assimilated zinc oxide nanoparticles (ZnO‐NPs) and hydroxyapatite (HAp) nanostructures as filler and hydrophobic polycaprolactone (PCL) as matrix by a solution‐based chemical method. The ceramic carrier‐like nanostructures ZnO and HAp were synthesized by the in situ precipitation method. The physicochemical characterization of the prepared polymer‐coated drug ceramic nanocomposite (FCZ‐ZnO‐HAp‐PCL) was achieved using Fourier transform infrared spectroscopy (FTIR), Field Emission Scanning Electron Microscopy (FESEM), particle size distribution (PSD), X‐ray diffraction (XRD), and Energy‐dispersive X‐ray (EDX) analysis. Furthermore, the biocompatibility and anticancer activity of the nanocomposite were explored by an MTT assay study. Successfully synthesized FCZ‐ZnO‐HAp‐PCL nanocomposite exhibited profound antimicrobial activity against targeted microbe species and potential cytotoxicity towards MCF‐7 human breast adenocarcinoma cell lines, which may be potentially used for the treatment of OM and prospective infections.

Abstract LB430: Identification of therapeutic target in breast cancer using G-quadruplex binding ligands

Abstract G-quadruplex (G4) motifs are noncanonical nucleic acid structures formed from staked guanine rich tetrads. G4 motifs have been shown to play significant effects on gene expression in all domains of life and are predominantly enriched in the regulatory regions such as promoters and enhancers of the human genome. G4-ligand mediated stabilization or destabilization of G4 structures has been discussed as a potential therapeutic strategy against tumor cells. However, the structure-specificity of G4-binding ligands remains a considerable challenge in determining the target-specificity. Therefore, it is necessary to understand the global effects of such G4-binding ligands at the transcriptome level to identify novel target(s) based on molecular structures and binding modes of both G4 motifs and ligands. In this study, we reported a top-down approach to extrapolate transcriptome-wide effects of two eminent G4-binding ligands Quarfloxin and Pyridostatin in human breast adenocarcinoma (MCF7) cell lines by RNA-seq analysis. Quadparser analysis was performed to analyze putative G4 forming sequences in protein coding breast cancer genes promoters. RNA-seq data was combined with Quadparser analysis to reveal that over 85% of the differentially expressed genes in both Pyridostatin and Quarfloxin contained G4 forming sequences in promoter region. Gene ontology analysis showed that both ligands affected key genes involved in extracellular matrix (ECM), architecture and cellular functions. A notable tumor suppressor deubiquitinase (DUBs) Cylindromatosis (CYLD) gene expression was particularly increased with Pyridostatin compared to Quarfloxin. In vitro assays also confirmed an increase in CYLD expression by qRT-PCR and Western blot. The detailed molecular mechanism of G4-mediated increase in CYLD expression is under study. Collectively, the current results present a new approach to identify G4-ligands binding target in breast cancer cells and provide insights into designing G4-ligands as an anti-cancer therapeutics. Citation Format: Maria Razzaq, Subramaniyam Ravichandran, Nazia Parveen, Kyeong Kyu Kim. Identification of therapeutic target in breast cancer using G-quadruplex binding ligands [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr LB430.

In Vitro Antiviral and Anticancer Effects of Tanacetum sinaicum Essential Oil on Human Cervical and Breast Cancer.

Cervical cancer has been linked to human papillomavirus (HPV) types 16 and 18. Essential oils (EOs) are vital natural products of plants with various therapeutic and biological properties. The purpose of this study is to investigate and assess Tanacetum sinaicum essential oil's possible antiviral and anticancer properties, with a focus on its in vitro effects on human cervical cancer and human breast adenocarcinoma cell lines. Tanacetum sinaicum EO was extracted via hydrodistillation (HD) and characterized using gas chromatography-mass spectrometry (GC-MS). MTT assay was used to determine the cell viability of Hela (a human epithelial cervical cancer) and MCF-7 (human breast adenocarcinoma) cell lines. Quantitative real-time polymerase chain reaction (PCR) was utilized to assess the antiviral efficacy of EO against HPV-16 and 18, and anti-metastatic characteristics. The biological activity of EO was assessed using Autophage and Cell genotoxicity via the comet assay. EO is mostly composed of chrysanthenyl acetate, thujone, and verbenol. The cell viability was reduced after 24 hours of incubation at doses from 100 to 400 µg/ml. Concentrations of 800 to 3,200 µg/ml significantly inhibit cell growth. After a 24-hour incubation period, doses ranging from 100 to 400 µg/ml reduced cell viability from 62 to 72%. Concentrations of 800 to 3,200 µg/ml significantly suppress cell growth by over 95%. In MCF7 and HeLa cell lines, EO lowered virus copy numbers in a dose-dependent manner, with higher concentrations of the oil inhibiting virus replication more effectively. EO treatment increased the number of autophagosomes/autolysosomes and acidic vesicular organelles in both cell lines. On the HeLa and MCF7 cell lines, EO demonstrated antiproliferative and antimetastatic effects. The results demonstrated that EO had dose-dependent genotoxic effects on both cancer cell lines, as evidenced by DNA damage. Tanacetum sinaicum EO is a prospective source of natural bioactive compounds that can be employed in pharmaceutical and medicinal applications due to its antiviral, antiproliferative, anti-metastatic and genotoxic properties.

Design, Synthesis and In-Silico Studies of Piperidine-Dihydropyridine Hybrids as Anticancer Agents.

In this study, we designed, synthesized and characterized a novel series of piperidine-dihydropyridine hybrid compounds and characterized them by 1H-NMR, 13C NMR, mass spectrometry (MS), and elemental analysis. Subsequently, we assessed their in vitro anticancer potentials against the human breast adenocarcinoma cell line MCF-7 and the lung cancer cell line A-549. Several of these compounds demonstrated significant activity, with IC50 values ranging from 15.94 μM to 48.04 μM for A-549 and 24.68 μM to 59.12 μM for MCF-7, when compared to the reference drug Cisplatin.Notably, a compound featuring a 3-fluoro substitution in the carboxamide series exhibited robust inhibitory effects, with an IC50 of 15.94±0.201 μM against A-549 cells and an IC50 of 22.12±0.213 μM against MCF-7 cells, respectively. Additionally, a compound containing a cyclobutyl ring displayed potent activity, with an IC50 of 16.56±0.125 μM against A-549 and an IC50 of 24.68±0.217 μM against MCF-7 cells, respectively. Furthermore, molecular docking studies against the Epidermal Growth Factor Receptor (EGFR) (PDB ID: 2J6M) revealed favourable binding scores and interactions, suggesting their potential as promising candidates for further investigation in the context of anticancer drug development.

Conjugation of an anti-metabolite nucleobase analogue with a mitochondriotropic agent via palladium(II) against breast cancer cells

The conjugation of the uracil (a nucleobase) analogue, 6-methyl-thiouracil (MTUC), with the mitochondriotropic agent of Tri-o-Tolyl-Phosphine (TOTP) through palladium(II) leads to the formation of the metallodrug of formula [PdCl(TOTP)(MTUC)] (1). The metallodrug was characterized in solid state using Attenuated Total Reflectance-Fourier Transform Infra-Red (ATR-FTIR) spectroscopy and X-ray diffraction crystallography (XRD), while its behavior in solution was examined through Ultra Violet (UV) and 1H NMR spectroscopies. The in vitro cytotoxicity of 1 was assessed against human breast adenocarcinoma cell lines: MCF-7 (hormone-dependent (HD)) and MDA-MB-231 (hormone-independent (HI)), as well as fetal lung fibroblast (MRC-5) cells. The MCF-7 cell morphology suggests apoptotic pathway, and this was confirmed by Acridine Orange/Ethidium Bromide (AO/EB) Staining, and the loss of the permeabilization of the mitochondrial membrane. The binding affinity of 1 toward the calf thymus (CT) DNA was clarified.

New Imidazo[4,5‐c]pyridine‐piperidine Hybrids as Potential Anti‐cancer Agents and In‐Silico Studies

AbstractDesign and synthesis of a series of novel hybrid molecules that combine Imidazo[4,5‐c] pyridines with piperdines are presented in this paper. Total 17 derivatives were meticulously synthesized and characterized using 1H NMR, 13C NMR, MS and elemental analysis techniques. The in vitroanticancer activities are estimated by verifying their effectiveness against the MCF‐7 human breast adenocarcinoma and A549 lung cancer cell line, with cisplatin and doxorubicin serving as reference drugs. Several of these compounds demonstrated significant potential, displaying IC50 values within the range of 12.26–84.5 μM for A549, and 13.37–69.82 μM for MCF‐7. Notably, compound 11 bis found to be more potent than the standard drug cisplatin with an IC50 of 12.26±0.8 μM against A549 cells, while compound 11 d exhibited highest inhibition with an IC50 of 13.37±0.4 μM against MCF‐7 cells. Although its effectiveness was moderately lower when compared to doxorubicin, it still retained substantial anticancer activity. Molecular docking studies were performed to decouple the binding affinity and ligand interactions of the compounds with the estrogen receptor alpha (ERα) (PDB ID: 3ERT). The pharmacokinetic evaluation revealed favourable drug‐likeness properties for all the molecules, suggesting their potential as therapeutic agents.

Photoinduced cytotoxic activity of a rare ruthenium nitrosyl phenanthroline complex showing NO generation in human cells.

A new nitro-nitrosyl complex [RuNO(Phen)(NO2)2OH] (1) was synthesized and characterized by X-ray diffraction, where Phen = 1,10-phenanthroline. The complex was crystallized in two different modifications without (1) and with a solvent molecule of DMF (1a). The photolysis process together with the determination of the quantum yield of NO release was investigated in acetonitrile solution using a special flow-through system for the simultaneous registration of infrared (IR) and optical absorption (UV-vis) spectra under irradiation with 450 nm light. The quantum yield of photoinduced NO release was 4.0 ± 0.2%. DFT calculations showed that the main contribution to the absorption band at 450 nm is made by the HOMO/HOMO-1 → LUMO transitions, which are represented by the transfer of electron density from the -OH and -NO2 ligands to the orbitals located on the Ru-NO bond. The dark and photoinduced cytotoxicity of the complex was studied against the human breast adenocarcinoma (MCF-7) and lung carcinoma (A549) cell lines and human non-tumor lung fibroblasts (MRC5). The complex shows a low cytotoxicity on MCF-7 cells (ICdark50 = 90.6 ± 6.2 μM and ICirr.50 = 95.3 ± 11.4 μM) and a moderate dark cytotoxicity on A549 and MRC5 cells (ICdark50 = 33.4 ± 2.6 μM and ICdark50 = 62.6 ± 3.1 μM, respectively), which slightly increases after irradiation (ICirr.50 = 21.2 ± 3.3 μM and ICirr.50 = 47.2 ± 2.3 μM, respectively).