3 Biotech最新文献

Triple-negative breast cancer (TNBC) is characterized by a diverse range of molecular features that have been extensively studied. MYC plays a critical role in regulating metabolism, differentiation, proliferation, cell growth, and apoptosis. Dysregulation of MYC is associated with poor prognosis and contributes to the development and progression of breast cancer. A particularly intriguing aspect of TNBC is its association with tumors in BRCA1 mutation carriers, especially in younger women. MYC may also contribute to resistance to adjuvant treatments. For TNBC, targeting MYC-regulated pathways in combination with inhibitors of other carcinogenic pathways offers a promising therapeutic approach. Several signaling pathways regulate TNBC, and targeting these pathways could lead to effective therapeutic strategies for breast cancer. Advances in genomic tools, such as CRISPR-Cas9, next-generation sequencing, and whole-exome sequencing, are revolutionizing breast cancer diagnoses. These technologies have significantly enhanced our understanding of MYC oncogenesis, particularly through CRISPR-Cas9 and NGS. Targeting MYC and its partner MAX could provide valuable insights into TNBC. Moreover, the therapeutic potential of targeting MYC-driven signaling mechanisms and their interactions with other oncogenic pathways, including PI3K/AKT/mTOR and Wnt/β-catenin, is increasingly recognized. Next-generation sequencing and CRISPR-Cas9 represent significant breakthroughs in genomic tools that open new opportunities to explore MYC's role in TNBC and facilitate the development of personalized treatment plans. This review discusses the future clinical applications of personalized treatment strategies for patients with TNBC.

The present investigation focused on exploring the potential of fungal endophytes as a valuable source of bioactive compounds with diverse applications. The phenolic compound 5-methylmellein was isolated for the first time from Alternaria burnsii, an endophytic fungus associated with Morus alba Linn. The compounds were structurally characterized using comprehensive spectral analysis, including ¹H-, ¹³C, and 2D-NMR, as well as HRESI-MS. The study investigated the antifungal activity of 5-methylmellein against several plant pathogenic fungi, including Botrytis cinerea, Colletotrichum gloeosporioides, Cercospora beticola, and Rhizoctonia solani. In vitro assays showed significant inhibition of various plant pathogenic fungi, and the IC₅₀ values ranging from 34.59 ± 1.03^a µg/mL to 44.76 ± 1.03^b µg/mL against the tested fungi. In vivo experiments on apples and grapes revealed that 5-Methylmellein significantly reduced fruit decay caused by Botrytis cinerea. The wound incidence in the control group reached 95.78%, while the treated groups exhibited a reduction of 37.54% after 15 days. These findings underscore the potential of 5-methylmellein as a potent antifungal agent, suggesting its eco-friendly application in agriculture for managing fruit decay.

This study investigated the potential of MET kinase inhibitors, cabozantinib, crizotinib, and PHA665752, in reversing multidrug resistance (MDR) mediated by ABCB1 in cancer cells. The accumulation of the fluorescent probe, Rhodamine 123, was assessed using flow cytometry and fluorescence microscopy in MDR MES-SA/DX5 and parental cells. The growth inhibitory activity of MET inhibitors as monotherapies and in combination with chemotherapeutic drugs was evaluated by MTT assay. CalcuSyn software was used to analyze the combination index (CI) as an index of drug-drug interaction in combination treatments. Results showed that at concentrations of 5, and 25 μM, c-MET inhibitors significantly increased Rhodamine 123 accumulation in MDR cells, with ratios up to 17.8 compared to control cells, while exhibiting no effect in parental cells. Additionally, the combination of c-MET inhibitors with the chemotherapeutic agent doxorubicin synergistically enhanced cytotoxicity in MDR cells, as evidenced by combination index (CI) values of 0.54 ± 0.08, 0.69 ± 0.1, and 0.85 ± 0.07 for cabozantinib, crizotinib, and PHA665752, respectively. While all three c-MET inhibitors stimulated ABCB1 ATPase activity in different manners at certain concentrations, PHA-665752 suppressed it at high concentration. In silico analysis also suggested that the transmembrane domains (TMD) of ABCB1 transporters could be considered potential target for these agents. Our results suggest that c-MET inhibitors can serve as promising MDR reversal agents in ABCB1-medicated drug-resistant cancer cells.

The overexpression of Kruppel-like factor 5 (KLF5) appears in several types of cancer. KLF5 may be an effective therapeutic target for treating OC, but its function in ovarian cancer (OC) remains unknown. The KLF5 mRNA expression levels in several OC cell lines were analyzed using RT-qPCR. Then, NC-siRNA or KLF5-siRNA was transfected into SK-OV-3 and OVCAR-3 cells. RT-qPCR and WB were used to detect the efficiency of KLF5 silence, CCK-8, colony formation assay, IHC staining, flow cytometry, and WB were performed to investigate the KLF5 function on OC cell proliferation and the activation of the extracellular signal-regulated Kinase (ERK)/mitogen-activated protein kinase (MAPK) signaling pathway. Next, a dual-luciferase and IF assay were used to determine the relationship between KLF5 and the Ras response element-binding protein (RREB1). SK-OV-3 and OVCAR-3 cells were treated with KLF5-siRNA and C16-PAF + EGF (MAPK agonist), separately or in combination. Proteins including KLF5, RREB1, p-p38, p-ERK1/2, ERK5, p-ERK5, Cyclin D1, CDK4, and CDK6 were quantified by WB. Finally, CCK-8, colony formation assay, and flow cytometry were employed again. KLF5 is highly expressed in OC cells compared with normal cells. When KLF5 knockdowns in SK-OV-3 and OVCAR-3 cells, the cell proliferation restrains, and the G1 phase prolongs. In addition, KLF5 silence caused a decrease of Cyclin D1, CDK4, CDK6, p-p38, p-ERK1/2, and p-ERK5/ERK5 expression levels. However, these statuses could be revised by C16-PAF + EGF. Results also found that when the ERK/MAPK signaling is activating, RREB1 is expressed low. The KLF5 silence could up-regulate the RREB1 expression. The KLF5 silence could restrain the OC cell proliferation and cell cycle. KLF5-siRNA may target upregulating RREB1 expression, thereby inhibiting the activation of the ERK/MAPK signaling pathway in OC cells.

We present the chemical synthesis of polyethyleneimine-conjugated silver sulfide nanoparticles (PEI/AS) utilizing an economical solvothermal synthesis method, aimed at developing effective alternative antibacterial agents. The antibacterial efficacy of the synthesized materials, both with and without the application of near-infrared (NIR) laser irradiation, was evaluated in vitro against two distinct clinically relevant multi-drug-resistant (MDR) uropathogenic strains: Escherichia coli and methicillin-resistant Staphylococcus aureus. The bactericidal effects induced by NIR light indicate that the PEI/AS nanoparticles possess an efficiency that is five times greater than that of Ag₂S alone. A suggested antibacterial mechanism posits that the wrapping of PEI increases electrostatic interactions, thereby facilitating the attachment of Ag₂S nanoparticles to the bacterial surface. This process leads to the disruption of the outer membrane through the generation of localized heat and an increased concentration of reactive oxygen species (ROS), including superoxide anions (·O₂ ^-) and hydroxyl radicals (·OH). In addition, the mechanism involves the regulated release of Ag⁺ ions when exposed to NIR light irradiation. The combined action led to an over 95.79% elimination of bacteria at a concentration as low as 50 μg mL^-1, which can be primarily ascribed to the regulated photothermal effect induced by 808 nm near-infrared light irradiation, demonstrating exceptional photothermal conversion efficiency. These results paves a way for manufacturing innovation in future.

Supplementary information: The online version contains supplementary material available at 10.1007/s13205-024-04168-3.

Surfactin lipopeptides (LPs) are a compelling class of biosurfactants with notable antimicrobial and anticancer properties. This study presents a novel approach by integrating bioinformatics tools to assess the drug potential of Surfactin, specifically focusing on its antibacterial, antifungal activities, and cancer cell-line toxicity. Silver nanoparticles (AgNPs) were synthesized using Surfactin, a biosurfactant derived from Bacillus subtilis KLP2016, as a capping agent, both in the presence and absence of Surfactin, to evaluate its impact on nanoparticle stability and bioactivity. The Surfactin-capped AgNPs demonstrated enhanced stability, uniformity, and antimicrobial efficacy, confirmed through UV-VIS spectroscopy, FE-SEM, and X-ray diffraction analysis. The bioinformatics approach, including ADMET and PASS analysis, revealed the potential of Surfactin as a potent antimicrobial and anticancer agent. In addition, molecular docking studies further validated the interaction of Surfactin with key microbial cell-wall enzymes and proteins, underscoring its therapeutic potential. These findings suggest that Surfactin-stabilized AgNPs, combined with bioinformatic predictions, could pave the way for innovative antimicrobial and anticancer therapies.

Rapamycin analogs are approved by the FDA for breast and renal cancer treatment. Hence, the possibility of nanoparticle-mediated delivery of Rapamycin could be examined. In the present study, PEGylated Gold-core shell iron oxide nanoparticles were used for the targeted delivery of Rapamycin, and R-Au-IONPs were formulated. SEM, XRD, and FTIR determined the smooth spherical morphology, and compositional structure, and confirmed the conjugation of Rapamycin onto the NPs. The in vitro drug release study showed a controlled release of the drug over time. R-Au-IONPs showed significant cytotoxicity in MCF 7 cells. Anti-proliferative assays such as trypan blue dye exclusion assay, microscopy, Fluorescent staining, and clonogenic assays were performed. NH staining, Rhodamine 123 staining, PS externalization, and the cleavage of PARP protein by western immunoblot assays confirmed the induction of apoptosis. The mechanism of R-Au-IONP-induced cell death was analyzed by flow cytometry. Our in-vitro study, on the impact of R-Au-IONPs on cell viability in the human breast adenocarcinoma cell line (MCF-7), confirms the efficacy of drug delivery using the nanoparticle system. Further results implied the induction of apoptosis. This drug delivery system using Rapamycin could be a potential candidate in the treatment of breast cancer.

Cardamom mosaic virus causing mosaic/katte disease is the most destructive virus infecting cardamom. The development of effective diagnostic assays is essential for the production of virus-free plants, as the primary spread of the virus occurs through vegetative propagation. Currently used PCR-based assays are not suitable for Point-of-Care testing, require sophisticated equipment, and are time-consuming. Hence, in the present study, an assay based on reverse transcription-recombinase polymerase amplification (RT-RPA) combined with lateral flow assay (RT-RPA-LFA) was optimized for the specific, and sensitive detection of CdMV. The forward and reverse primers selected for RT-RPA were labeled with 6-carboxyfluorescein (FAM) and biotin respectively at the 5´end. The tedious total RNA preparation was avoided by using the crude extract as a template for the assay. A magnesium acetate concentration of 14 mM, 0.4 M betaine, temperature from 37 to 42 ℃, and 20 min of incubation time were found optimum for the assay. The entire RT-RPA-LFA from sample preparation to visualization of results could be completed within 40-50 min and the assay is suitable for Point-of-Care testing. The assay is specific for CdMV and could detect the virus up to 10^-5 dilutions of the crude extract. The assay was validated using field samples collected from different cardamom-growing regions of Kerala and Karnataka, India.

Supplementary information: The online version contains supplementary material available at 10.1007/s13205-024-04191-4.

We have developed novel and sustainable homogeneous catalysts employing Glutamic acid (Glu) as a biodegradable and eco-friendly organocatalyst for the synthesis of N-(4-oxo-2-phenyl-1,2-dihydroquinazolin-3(4H)-yl)isonicotinamide derivatives (5a-l) via multicomponent reactions (MCRs) of isatoic anhydride, isoniazid and heteroaromatic/aromatic aldehyde in ethanol on oil bath stirring at 60 °C. Selected final product homogeneity was examined by various spectroscopic techniques such as ¹³C-, ¹H- NMR, FT-IR and LC-MS. For the first time, herein investigated electrochemical behavior of selected derivatives (5c, 5h-l) using cyclic voltammetry method. The results of this investigation indicated compounds 5i, 5h and 5l exhibited highest levels of oxidation and reduction potential. Further, pharmacokinetic properties were assessed via SwissADME online tool, derivatives tested complied with Lipinski's rule of five for drug-likeness. Furthermore, molecular docking studies demonstrated for significant binding between the protein and ligand, and affinity values ranged from - 8.91 to - 8.45 kcal/mol, and MM/PBSA estimated high negative values suggested significant interactions between ligand and protein. Moreover, antibacterial evaluation of compounds 5i in water, and 5k in DMSO on Salmonella typhimurium showed pronounced effect with inhibition zone of 15 ± 0.6 mm and 20 ± 0.4 mm, respectively as compared to the standard tetracycline inhibition zone of 15 ± 0.5 mm.

Supplementary information: The online version contains supplementary material available at 10.1007/s13205-024-04188-z.

CRISPR-Cas9 (clustered regularly interspaced short palindromic repeats-associated protein 9) has revolutionized gene editing tools and paved the way for innovations in medical research for disease diagnosis and treatment. However, better specificity and efficient delivery of this gene machinery make it challenging to successfully edit genes for treating various diseases. This is mainly due to cellular barriers, instability in biological environments, and various off-target effects that prohibit safe and efficient delivery under in vivo conditions. This review examines several delivery modes [plasmid, mRNA, RNP (ribonucleoprotein)] and methods for the CRISPR-Cas9 system delivery, focusing on its potential applications in cancer therapy. Biocompatibility and cytotoxicity are crucial factors determining their safe and effective use. Various nanomaterials have been reviewed for their biocompatibility, limitations, and challenges in treating cancer. Among the reviewed nanoparticles, lipid nanoparticles (LNPs) stand out for their biocompatibility due to their biomimetic lipid bilayer that effectively delivers CRISPR/Cas9 cargoes while reducing toxicity. We discuss challenges in in vivo delivery and associated findings such as encapsulation, target delivery, controlled release, and endosomal escape. Future directions involve addressing limitations and adapting CRISPR-Cas9 for clinical trials, ensuring its safe and effective use.