HeLa Research Articles

Investigating the impact of thiol reactivity and disulfide formation on cellular uptake of cell-permeable peptides.

Cell-penetrating peptides (CPPs) have been explored as versatile tools to transport various molecules into cells. The uptake mechanism of CPPs is still not clearly understood and most probably depends on several factors like the nature of the CPP itself, the attached cargo, the investigated cell system, and other experimental conditions, such as temperature and concentration. One of the first steps of internalization involves the interaction of CPPs with negatively charged molecules present at the outer layer of the cell membrane. Recently, thiol-mediated uptake has been found to support the effective translocation of sulfhydryl-bearing substances that would actually not be cell-permeable. Within this work, we aimed to understand the relevance of thiol reactivity for the uptake mechanism of cysteine-containing CPPs that we have developed previously in our group. Therefore, we compared the two peptides, sC18-Cys and CaaX-1, in their single reduced and dimeric disulfide versions. Cytotoxicity, intracellular accumulation, and impact on the internalization process of the disulfides were investigated in HeLa cells. Both disulfide CPPs demonstrated significantly stronger cytotoxic effects and membrane activity compared with their reduced counterparts. Notably, thiol-mediated uptake could be excluded as a main driver for translocation, showing that peptides like CaaX-1 are most likely taken up by other mechanisms.

Açaí (Euterpe oleracea Mart.) Seed Oil and Its Nanoemulsion: Chemical Characterisation, Toxicity Evaluation, Antioxidant and Anticancer Activities.

This study explores a nanoemulsion formulated with açaí seed oil, known for its rich fatty acid composition and diverse biological activities. This study aimed to characterise a nanoemulsion formulated with açaí seed oil and explore its cytotoxic effects on HeLa and SiHa cervical cancer cell lines, alongside assessing its antioxidant and toxicity properties both in vitro and in vivo. Extracted from fruits sourced in Brazil, the oil underwent thorough chemical characterization using gas chromatography-mass spectrometry. The resulting nanoemulsion was prepared and evaluated for stability, particle size, and antioxidant properties. The nanoemulsion exhibited translucency, fluidity, and stability post centrifugation and temperature tests, with a droplet size of 238.37, PDI -9.59, pH 7, and turbidity 0.267. In vitro assessments on cervical cancer cell lines revealed antitumour effects, including inhibition of cell proliferation, migration, and colony formation. Toxicity tests conducted in cell cultures and female Swiss mice demonstrated no adverse effects of both açaí seed oil and nanoemulsion. Overall, açaí seed oil, particularly when formulated into a nanoemulsion, presents potential for cancer treatment due to its bioactive properties and safety profile.

Quantification of Intracellular DNA-Protein Cross-Links with N7-Methyl-2'-Deoxyguanosine and Their Contribution to Cytotoxicity.

The major product of DNA-methylating agents, N7-methyl-2'-deoxyguanosine (MdG), is a persistent lesion in vivo, but it is not believed to have a large direct physiological impact. However, MdG reacts with histone proteins to form reversible DNA-protein cross-links (DPCMdG), a family of DNA lesions that can significantly threaten cell survival. In this paper, we developed a tandem mass spectrometry method for quantifying the amounts of MdG and DPCMdG in nuclear DNA by taking advantage of their chemical lability and the concurrent release of N7-methylguanine. Using this method, we determined that DPCMdG is formed in less than 1% yield based upon the levels of MdG in methyl methanesulfonate (MMS)-treated HeLa cells. Despite its low chemical yield, DPCMdG contributes to MMS cytotoxicity. Consequently, cells that lack efficient DPC repair by the DPC protease SPRTN are hypersensitive to MMS. This investigation shows that the downstream chemical and biochemical effects of initially formed DNA damage can have significant biological consequences. With respect to MdG formation, the initial DNA lesion is only the beginning.

A novel benzothiazole-based fluorescent probe for continuous detection of Cu2+/S2− and its application in cell imaging

In this paper, a fluorescent probe methyl(E)-2-(3-(benzo[d]thiazol-2-yl)-2‑hydroxy -5-methylbenzylidene) hydrazine-1-carboxylate (DYH) based on benzothiazole was designed and synthesized. The characterization involved NMR, HRMS, and crystal structure analysis. In case of the existence of other competing cations, the probe DYH showed high-affinity for Cu2+, and the resultant DYHCu2+ showed exceptional capacity to recognize S2− in CH3CN/HEPES (7/3, v/v, HEPES 0.1 M, pH = 7.3). The detection limits of DYH for Cu2+ and DYHCu2+ for S2− were 0.56 μM and 0.67 μM, respectively. It was determined that the binding stoichiometry between the DYH and Cu2+, as well as DYHCu2+ and S2−, was 1:1 according to the job's curve. The pH of the probe and complex was stable between 4 and 10. Colorimetric experiments of the probe DYH were conducted using test paper as a carrier. Additionally, DYH was used for fluorescence imaging of Cu2+ and S2− in Hela cells with poor cellular toxicity and good cell permeability.

Antimicrobial efficacy of Punica granatum Lythraceae peel extract against pathogens belonging to the ESKAPE group.

The improper use and abuse of antibiotics have led to an increase in multidrug-resistant (MDR) bacteria resulting in a failure of standard antibiotic therapies. To date, this phenomenon represents a leading public health threat of the 21st century which requires alternative strategies to fight infections such as the identification of new molecules active against MDR strains. In the last 20 years, natural extracts with biological activities attracted scientific interest. Following the One Health Approach, natural by-products represent a sustainable and promising alternative solution. Consistently, the aim of the present study was to evaluate the antimicrobial activity of hydro-alcoholic pomegranate peel extract (PPE) against MDR microorganisms belonging to Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter spp. "ESKAPE" group pathogens. Through semiquantitative and quantitative methods, the PPE showed effective antimicrobial activity against Gram-positive and Gram-negative MDR bacteria. The kinetics of bactericidal action of PPE highlighted that microbial death was achieved in a time- and dose-dependent manner. High concentrations of PPE exhibited antioxidant activity, providing a protective effect on cellular systems and red blood cell membranes. Finally, we report, for the first time, a significant intracellular antibacterial property of PPE as highlighted by its bactericidal action against the staphylococcal reference strain and its bacteriostatic effect against clinical resistant strain in the HeLa cell line. In conclusion, due to its characterized content of polyphenolic compounds and antioxidant activity strength, the PPE could be considered as a therapeutic agent alone or in conjunction with standard antibiotics against challenging infections caused by ESKAPE pathogens.

Multiple Fano resonator based on photonic crystal waveguide coupled with two micro-cavities for biomedical sensing application

In order to acquire a miniature refractive index (RI) biosensor with high sensitivity, fast and selective for ultra-low concentrations of molecules, a new Photonic Crystal (PhC) biosensor based on a waveguide coupled to a Fano resonator is proposed for cancer cells detection. An optimized structure of the biosensor can detect cells cancers (Besal, Hela, Jurkat and PC12) in a biological solution deposited on the surface of the resonator. The detection mechanism uses the refractive index as a detection element. The performance of the proposed biosensor is studied by analyzing the variations in the transmission spectrum of different normal and cancer cells. The proposed structure is multimode PhC, with silica as a dielectric material. The finite element method (FEM) have been implemented for studying and investigating the numerical values. The simulation results display that the proposed biosensor attains spectral sensitivities of ‘513.12 nm RIU−1’, ‘587.28 nm RIU−1 ’, ‘702.35 nm RIU−1 ’ and ‘690.57 nm RIU−1 ’ corresponding to Hela cells, PC12 cells, Basal cells and Jurkat cells, respectively. And he qualilty factor Q of the Fano resonance mode can reach 3040.26. Our optimized design ensures easy fabrication with ongoing techniques. This study may open a new way for the development of integrated optical circuits and biosensing.

Methotrexate-loaded manganese nitrogen dual-doped carbon quantum dots as targeted nano drug-delivery system for potential use in cancer theranostics

The present work highlights manganese nitrogen dual-doped carbon quantum dots as a novel pH-sensitive drug delivery system, for the delivery of methotrexate, targeted to cancer cells, along with the simultaneous real-time monitoring of cellular uptake and drug delivery using fluorescence imaging. Briefly, methotrexate was conjugated with the as-prepared quantum dots to form a nano drug-delivery system through non-covalent interactions. The developed material demonstrated drug loading efficiency of ∼91 % and a very efficient (∼80 %) pH-responsive drug release at pH 5.5. The cytotoxic properties of the developed nano-platform viz. manganese, nitrogen dual-doped carbon quantum dots, and methotrexate conjugated manganese nitrogen dual-doped carbon quantum dot complex were evaluated in cancer cells (HeLa) and healthy human cells (L929). The conjugated complex demonstrated excellent biocompatibility, high cellular uptake, and a substantial drug release. Healthy human cells were largely unaffected when incubated with both the MTX-conjugated Mn-, N-CQD and the free drug. Additionally, in vitro, the fluorescence bio-imaging potential of the developed nano-platform was also assessed, where the unconjugated (free) quantum dots exhibited better blue fluorescence imaging potential compared to that of the conjugated complex, which can be attributed to fluorescence quenching due to the presence of methotrexate. This study highlights the manganese nitrogen dual-doped carbon quantum dots capable of pH-responsive methotrexate drug delivery selective to cancer cells. The combination of drug delivery and fluorescence imaging creates an efficient platform for targeted therapy, allowing very precise drug release while accessing non-invasive monitoring of therapeutic response. This method holds a great promise for advancing cancer treatment strategies, with the potential for increased efficacy and decreased side effects.

Ionic liquid based ultrasonic assisted extraction of flavonoids from Artemisia vulgaris and in vitro evaluation of anticancer activity

This study was focused on an effective technique of flavonoids extraction from Artemesia vulgaris using ionic liquids (ILs) as green solvents. The findings showed that the hydrophilic IL, [Bmim][Br] coupled with ultrasonic-assisted extraction (UAE) was able to extract flavonoids better than hydrophobic IL, [Bmim][PF6]. Single-factor experiments reported that the optimum extraction time, 30 min and solid-to-liquid ratio, 1:10 could obtain high extraction yield (9.3 %) and flavonoid content (5.30 mg/g). With the assistance of ultrasonic energy, [Bmim][Br] was successfully to lysis cells and leach out flavonoids as explained by the morphological observation of SEM and functional groups in FTIR spectrum. UHPLC-MS/MS detected few intense flavonoids such as rutin, kaempferol-3-rutinoside, and quercetin-3-glucopyranoside in the extract. The monitoring of Kamlet-Taft parameters such as polarity, acidity and basicity explained the formation of hydrogen bonding between IL, cellulose and flavonoids. The extraction of flavonoids in [Bmim][Br] based UAE followed Peleg’s kinetic model satisfactorily (R2 > 0.95). [Bmim][Br] could be reused up to 5 cycles before lost its extractability. The recovered flavonoids extract exhibited high anticancer activity against HeLa cells (IC50 = 347.37 µg/mL) without any significant cytotoxic effect on normal cervical cells at the concentration up to 300 µg/L. Therefore, this technique was likely to be an efficient extraction of plant flavonoids.

Nitroxyl donating and visualization with a coumarin-based fluorescence probe

Nitroxyl (HNO), the single-electron reduction product of nitric oxide (NO), has attracted great interest in the treatment of congestive heart failure in clinical trials. In this paper, we describe the first coumarin-based compound N-hydroxy-2-oxo-2H-chromene-6-sulfonamide (CD1) as a dualfunctional HNO donor, which can release both an HNO signaling molecule and a fluorescent reporter. Under physiological conditions (pH 7.4 and 37 °C), the CD1 HNO donor can readily decompose with a half-life of ∼90 min. The corresponding stoichiometry HNO from the CD1 donor was confirmed using both Vitamin B12 and phosphine compound traps. In addition to HNO releasing, specifically, the degradation product 2-oxo-2H-chromene-6-sulfinate (CS1) was generated as a fluorescent marker during the decomposition. Therefore, the HNO amount released in situ can be accurately monitored through fluorescence generation. As compared to the CD1 donor, the fluorescence intensity increased by about 4.9-fold. The concentration limit of detection of HNO releasing was determined to be ∼0.13 μM according to the fluorescence generation of CS1 at physiological conditions. Moreover, the bioimaging of the CD1 donor was demonstrated in the cell culture of HeLa cells, where the intracellular fluorescence signals were observed, inferring the site of HNO release. Finally, we anticipate that this novel coumarin-based CD1 donor opens a new platform for exploring the biology of HNO.

Cyclopropenes as Chemical Reporters for Dual Bioorthogonal and Orthogonal Metabolic Labeling of DNA.

Dual bioorthogonal labeling enables the investigation and understanding of interactions in the biological environment that are not accessible by a single label. However, applying two bioorthogonal reactions in the same environment remains challenging due to cross-reactivity. We developed a pair of differently modified 2'-deoxynucleosides that solved this issue for dual and orthogonal labeling of DNA. Inverse-electron demand Diels-Alder and photoclick reactions were combined to attach two different fluorogenic labels to genomic DNA in cells. Using a small synthetic library of 1- and 3-methylcyclopropenyl-modified 2'-deoxynucleosides, two 2'-deoxyuridines were identified to be the fastest-reacting ones for each of the two bioorthogonal reactions. Their orthogonal reactivity could be evidenced in vitro. Primer extension experiments were performed with both 2'-deoxyuridines investigating their replication properties as substitutes for thymidine and evaluating subsequent labeling reactions on the DNA level. Finally, dual, orthogonal and metabolic fluorescent labeling of genomic DNA was demonstrated in HeLa cells. An experimental procedure was developed combining intracellular transport and metabolic DNA incorporation of the two 2'-deoxyuridines with the subsequent dual bioorthogonal labeling using a fluorogenic cyanine-styryl tetrazine and a fluorogenic pyrene-tetrazole. These results are fundamental for advanced metabolic labeling strategies for nucleic acids in the future, especially for live cell experiments.

Controlled Release of the Anticancer Drug Cyclophosphamide from a Superparamagnetic β-Cyclodextrin Nanosponge by Local Hyperthermia Generated by an Alternating Magnetic Field.

A β-cyclodextrin (β-CD) nanosponge (NS) was synthesized using diphenyl carbonate (DPC) as a cross-linker to encapsulate the antitumor drug cyclophosphamide (CYC), thus obtaining the NSs-CYC system. The formulation was then associated with magnetite nanoparticles (MNPs) to develop the MNPs-NSs-CYC ternary system. The formulations mentioned above were characterized to confirm the deposition of the MNPs onto the organic matrix and that the superparamagnetic nature of the MNPs was preserved upon association. The association of the MNPs with the NSs-drug complex was confirmed through field emission scanning electron microscopy, energy dispersive spectroscopy, transmission electron microscopy, X-ray photoelectron spectroscopy, dynamic light scattering, ζ-potential, atomic absorption spectroscopy, X-ray powder diffraction, selected area electron diffraction, and vibrating-sample magnetometer. The superparamagnetic properties of the ternary system allowed the release of CYC by utilizing magnetic hyperthermia upon the exposure of an alternating magnetic field (AMF). The drug release experiments were carried out at different frequencies and intensities of the magnetic field, complying with the "Atkinson-Brezovich criterion". The assays in AMF showed the feasibility of release by controlling hyperthermia of the drug, finding that the most efficient conditions were F = 280 kHz, H = 15 mT, and a concentration of MNPs of 5 mg/mL. CYC release was temperature-dependent, facilitated by local heat generation through magnetic hyperthermia. This phenomenon was confirmed by DFT calculations. Furthermore, the ternary systems outperformed the formulations without MNPs regarding the amount of released drug. The MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) assays demonstrated that including CYC within the magnetic NS cavities reduced the effects on mitochondrial activity compared to those observed with the free drug. Finally, the magnetic hyperthermia assays showed that the tertiary system allows the generation of apoptosis in HeLa cells, demonstrating that the MNPs embedded maintain their properties to generate hyperthermia. These results suggest that using NSs associated with MNPs could be a potential tool for a controlled drug delivery in tumor therapy since the materials are efficient and potentially nontoxic.

Cyclopropene als Chemische Reporter für die Duale Bioorthogonale und Orthogonale Metabolische Markierung von DNA

AbstractDie duale bioorthogonale Markierung ermöglicht die Untersuchung und das Verständnis von Wechselwirkungen in der biologischen Umgebung, die mit einer einzigen Markierung nicht zugänglich sind. Eine Herausforderung bleibt jedoch die Anwendung von zwei bioorthogonalen Reaktionen in der gleichen Umgebung aufgrund von Kreuzreaktivität. Um dieses Problem für die duale und orthogonale Markierung von DNA zu lösen, haben wir zwei unterschiedlich modifizierte 2′‐Desoxynukleoside entwickelt. Mit Hilfe der Diels‐Alder‐Reaktion mit inversem Elektronenbedarf und Photoklick‐Reaktion wurde genomische DNA in Zellen mit zwei verschiedenen fluorogenen Markern verknüpft. Aus einer kleinen synthetischen Bibliothek aus 1‐ und 3‐methylcyclopropenyl‐modifizierten 2′‐Desoxynukleosiden wurden zwei 2′‐Desoxyuridine identifiziert, die am schnellsten in jeweils einer der beiden bioorthogonalen Reaktionen reagieren. Ihre orthogonale Reaktivität konnte in vitro nachgewiesen werden. Ihre Replikationseigenschaften als Ersatz für Thymidin wurden in Primerverlängerungsexperimenten untersucht, um die anschließenden Markierungsreaktionen an DNA abzuschätzen. Schließlich wurde eine duale, orthogonale und metabolische Fluoreszenzmarkierung von genomischer DNA in HeLa‐Zellen nachgewiesen. Es wurde ein experimentelles Protokoll entwickelt, das den intrazellulären Transport und den metabolischen DNA‐Einbau der beiden 2′‐Desoxyuridine mit der anschließenden dualen bioorthogonalen Markierung unter Verwendung eines fluorogenen Cyanin‐Styryl‐Tetrazins und eines fluorogenen Pyren‐Tetrazols kombiniert. Diese Ergebnisse sind von grundlegender Bedeutung für fortgeschrittene metabolische Markierungsstrategien für Nukleinsäuren in der Zukunft, insbesondere in lebenden Zellen.

Metabolite Profiling, Biological and Molecular Analyses Validate the Nutraceutical Potential of Green Seaweed Acrosiphoniaorientalis for Human Health.

Seaweeds have proven to be nutrient-dense and are rich in antioxidants, like phenolics, flavonoids, and other essential metabolites that help to provide their medicinal benefits. Non-targeted metabolite profiling of the tropical green seaweed Acrosiphonia orientalis showed the presence of numerous groups of contents, including sugars, essential amino acids, and fatty acids. Targeted metabolite profiling using HPLC identified 17 amino acids. The extract exhibited a very low half-maximal effective concentration (EC50) dosage for HeLa and Huh-7 cell lines, indicating a high likelihood of anticancer properties. A significant positive correlation was found between biological activities, such as antioxidation, scavenging, and reducing power with the phenolic and flavonoid contents. The extract revealed augmentation of proliferation in selected cervical cells, as it upregulated p53 1.3-fold, and downregulated important cancerous genes such as Cas-3 and DNMT 12- and 8-fold, respectively. An approximate 55-fold downregulation was observed in selected hepatic cell lines. Microarray analysis of hepatic cells indicated 0.27% and 0.07% upregulation of coding and non-coding genes, respectively, and 0.41% and 0.13% downregulation of coding and non-coding genes, respectively. As a consequence, it can be said that A. orientalis has possible medicinal use, such as anticancer activity, and therefore may be an intriguing food component that has potential as a regular dietary supplement.

Protocol for Testing the Effects of ssDNA Aptamer in HeLa and MCF-7.

Conventional approaches for treating tumors encompass chemotherapy, radiotherapy, and surgery. However, these methods come with their limitations when applied in clinical practice. Aptamers are often referred to as "chemical antibodies" and consist of short DNA or RNA molecules, designed to bind to a wide range of targets, including proteins or nucleic acid structures. They exhibit strong affinities and remarkable specificity for their target molecules, making them capable of functioning as therapeutic agents to directly impede tumor cell proliferation. This approach helps minimize the harm to normal cells, thus reducing toxicity through decreased side effects. Here we report the procedure to develop ssDNA aptamer and investigate its ability to inhibit cancer cell proliferation in HeLa and MCF-7 cancer cell lines.

Exploring MiR-484 Regulation by Polyalthia longifolia: A Promising Biomarker and Therapeutic Target in Cervical Cancer through Integrated Bioinformatics and an In Vitro Analysis.

MiR-484, implicated in various carcinomas, holds promise as a prognostic marker, yet its relevance to cervical cancer (CC) remains unclear. Our prior study demonstrated the Polyalthia longifolia downregulation of miR-484, inhibiting HeLa cells. This study investigates miR-484's potential as a biomarker and therapeutic target in CC through integrated bioinformatics and an in vitro analysis. MiR-484 levels were analyzed across cancers, including CC, from The Cancer Genome Atlas. The limma R package identified differentially expressed genes (DEGs) between high- and low-miR-484 CC cohorts. We assessed biological functions, tumor microenvironment (TME), immunotherapy, stemness, hypoxia, RNA methylation, and chemosensitivity differences. Prognostic genes relevant to miR-484 were identified through Cox regression and Kaplan-Meier analyses, and a prognostic model was captured via multivariate Cox regression. Single-cell RNA sequencing determined cell populations related to prognostic genes. qRT-PCR validated key genes, and the miR-484 effect on CC proliferation was assessed via an MTT assay. MiR-484 was upregulated in most tumors, including CC, with DEGs enriched in skin development, PI3K signaling, and immune processes. High miR-484 expression correlated with specific immune cell infiltration, hypoxia, and drug sensitivity. Prognostic genes identified were predominantly epidermal and stratified patients with CC into risk groups, with the low-risk group showing enhanced survival and immunotherapeutic responses. qRT-PCR confirmed FGFR3 upregulation in CC cells, and an miR-484 mimic reversed the P. longifolia inhibitory effect on HeLa proliferation. MiR-484 plays a crucial role in the CC progression and prognosis, suggesting its potential as a biomarker for targeted therapy.

Exploring the antimicrobial and cytotoxic potential of novel chloroquine analogues.

Aim: To synthesize novel chloroquine analogues and evaluate them for antimicrobial and cytotoxic potential. Methods: Novel analogues were synthesized from chloroquine by nucleophilic substitution reaction at the 4-amino position. Results: Analogue CS1 showed maximum antimicrobial potential (30.3±0.15mm zone) against Pseudomonas aeruginosa and produced a 19.2±0.21mm zone against Candida albicans, while CS0 produced no zone at the same concentration. Analogue CS9 has excellent cytotoxic potential (HeLa cell line), showing 100% inhibition (IC50=8.9±1.2μg/ml), compared with CS0 (61.9% inhibition at 30μg/ml). Conclusion: These synthesized chloroquine analogues have excellent activity against different microbial strains and cervical cancer cell lines (HeLa) compared with their parent molecule.

Probing the Effects of Retinoblastoma Binding Protein 6 (RBBP6) Knockdown on the Sensitivity of Cisplatin in Cervical Cancer Cells.

Cervical cancer is a major cause of death in women despite the advancement of current treatment modalities. The conventional therapeutic agent, cisplatin (CCDP), is the standard treatment for CC; however, resistance often develops due to the cancer's heterogeneity. Therefore, a detailed elucidation of the specific molecular mechanisms driving CC is crucial for the development of targeted therapeutic strategies. Retinoblastoma binding protein 6 (RBBP6) is a potential biomarker associated with cell proliferation and is upregulated in cervical cancer sites, exhibiting apoptosis and dysregulated p53 expression. Furthermore, RBBP6 has been demonstrated to sensitize cancer cells to radiation and certain chemotherapeutic agents by regulating the Bcl-2 gene, thus suggesting a crosstalk among RBBP6/p53/BCL-2 oncogenic signatures. The present study, therefore, investigated the relationship between cisplatin and RBBP6 expression in CC cells. Herein, we first explored bioinformatics simulations and identified that the RBBP6/p53/BCL-2 signaling pathway is overexpressed and correlated with CC. For further analysis, we explored the Genomics of Drug Sensitivity in Cancer (GDSC) and found that most of the CC cell lines are sensitive to CCDP. To validate these findings, RBBP6 was silenced in HeLa and Vero cells using RNAi technology, followed by measurement of wild-type p53 and Bcl-2 at the mRNA level using qPCR. Cells co-treated with cisplatin and siRBBP6 were subsequently analyzed for apoptosis induction and real-time growth monitoring using flow cytometry and the xCELLigence system, respectively. Cancer cells in the co-treatment group showed a reduction in apoptosis compared to the cisplatin-treated group. Moreover, the real-time growth monitoring revealed a reduced growth rate in RBBP6 knockdown cells treated with cisplatin. Although wild-type p53 remained unchanged in the co-treatment group of cancer cells, Bcl-2 was completely repressed, suggesting that RBBP6 is necessary for sensitizing cervical cancer cells to cisplatin treatment by downregulating Bcl-2. The Vero cell population, which served as a non-cancerous control cell line in this study, remained viable following treatment with both siRBBP6 and cisplatin. Findings from this study suggest that RBBP6 expression promotes cisplatin sensitivity in HeLa cells through Bcl-2 downregulation. Knockdown of RBBP6 limits apoptosis induction and delays cell growth inhibition in response to cisplatin. The knowledge obtained here has the potential to help improve cisplatin efficacy through personalized administration based on the expression profile of RBBP6 among individual patients.

Two new triterpenoid saponins and two new goodyerosides isolated from the whole plant of Tournefortia sibirica

Two new triterpenoid saponins (oleanolic acid 2β-hydroxyl-3-O-β-D-glucuronopyranoside-6’-O-buthyl ester (1) and oleanolic acid 2β-hydroxyl-3-O-[β-D-glucuronopyranosyl-6′-O-methylester]-28-O-β-D-glucopyranoside (2)) and two new goodyerosides (4-methylenefuran-2(5H)-one (6′-O-vanilloyl)-β-D-glucopyranoside (3), 3-hydroxy-2(5H)-furanone, 4-(6′-O-vanilloyl)-β-D-glucopyranoside (4)), together with seven known compounds (5–11) were isolated from the whole plant of Tournefortia sibirica L. The chemical structures of the compounds were determined by spectroscopic analysis (1D and 2D NMR) and HR-ESI-MS. Compounds 1, 6 and 9 showed significant cytotoxicity towards A549, SK-Hep1 and HeLa cells, with IC50 values ranging from 1.68 ± 0.09 to 6.87 ± 0.13 μM.

Molecular modelling, cytotoxicity & biological investigation of novel fluorinated diphenylamine chalcone derivatives

A unique series of 1-(3-(3,4-difluorophenyl)amino)phenyl)ethan-1-one linked fluorinated diphenylamine chalcones (A1-A7) were synthesized and characterized by NMR, IR, and MS. The compounds were studied in vitro for antibacterial and antifungal activities. In vitro studies show that the compound had significant antibacterial (MICs 25–250 µg/ml) and antifungal (MICs 250–1000 µg/ml) properties. Chemicals A1, A2, and A4 significantly inhibited all bacterial strains, whereas compounds A1, A4, and A7 inhibited fungal very marginally. Compounds A1 and A2 had a high anti-Plasmodium falciparum activity. A molecular docking investigation discovered that compounds A1 and A2 significantly bind to the active site of falcilysin. Compound A1 maintains its stability through numerous van der Waals interactions with falcilysin protein residues. Compound A2, on the other hand, shows a hydrogen bond with Asn421, the secondary amine linked to the phenyl ring. The Molecular Dynamic simulation at 100 ns to investigate the dynamic stability of protein-ligand complexes. Compounds A1 and A2 exhibited the highest binding affinity and interaction. According to the MD simulation results, the proposed drug maintains the molecular interaction and structural integrity with P. falciparum. In HeLa cells, the cytotoxicity of compounds A1 and A4 was examined. A1 and A4 had computed LC50 values of 19.14 µg and 21.01 µg, respectively. Both compounds are cytotoxic to HeLa cells, according to the observations above. Compounds A1 and A2 are potential lead compounds for novel drug-like molecules having antibacterial, antifungal, and antimalarial properties.

An ESIPT active coumarin-diphenyl azine-based AIEgen: Nanomolar Cu2+ ion sensing, Latent Fingerprinting, live-cell imaging, and real sample analysis

Designing economical and sensitive chemosensors for detecting important metal ions, like Cu2+ ions, is essential and remains a hot topic worldwide. In this investigation, we extended our investigation to design a specific diphenyl-azine-based skeletal system for the selective and sensitive detection of Cu2+ions. We reported a coumarin-diphenyl-azine-based AIEgen (C-1) using an improvised methodology with a large Stoke's shift (∼245 nm) for precisely sensing copper ions in solid, solution, and biological cells. The AIE-ESIPT properties of C-1 were validated using various sophisticated techniques. C-1 was found to be a promising selective and sensitive sensor of Cu2+ ions (nanomolar LOD value ∼10.38 nM) via colorimetric and TURNOFF fluorometric ways. The practical applicability of C-1 was analyzed via real-time estimation of copper ions in water samples (obtained from three varied locations) and in biological samples (urine and serum). The satisfactory recovery with a low RSD value indicates the potential of C-1 in detecting copper ions in actual samples. Estimating copper ions in urine and blood samples demonstrates the usefulness of C-1 in diagnosing disorders like Wilson's disease, which is caused by high concentrations of copper ions in the body. Additionally, we have detected Cu2+ions in live cells (HeLa cells) and carried out Latent Fingerprinting (Level 1 and Level 2) to validate its versatility in physiological and forensic applications. Overall, the proposed C-1 AIEgen is an ultrasensitive Cu2+ions sensor with innumerable applications in the physical world.