A549 Cells Research Articles

Monotropein represses the proliferation and angiogenesis via the AKT/NF-κB pathway in lung cancer.

Monotropein (Mon) is an iridoid glycosides extracted from Morinda officinalis F.C. How. (Rubiaceae) that shows anticancer effects on colorectal cancer. This study aimed to investigate the therapeutic effect of Mon on lung cancer. The viability, apoptosis, invasion, and tube formation of A549 and H1299 were determined by CCK8 assay, flow cytometry, transwell assay, and tube formation assay. NF-κB expression in the nucleus was detected by immunofluorescent staining. In vivo assay, BALB/c nude mice were divided into a sham group and a 2mg/kg Mon group. For the Mon group, mice were orally administered with 2mg/kg Mon. The duration of the animal study was 35days, and the tumor formation and angiogenesis were investigated. Compared with the control (Mon 0μM) group, Mon dramatically repressed the viability, invasion, tube formation, proliferation marker Ki67, N-cadherin, and VEGFA expressions in A549 cells (Mon of 25, 50, and 100µM) and H1299 cells (Mon of 50 and 100µM). Mon dramatically promoted cell apoptosis, cleaved caspase 3, and E-cadherin expressions. Besides, Mon remarkably downregulated p-AKT and p-NF-κB protein expressions. Moreover, AKT agonist SC79 reversed the anticancer effects of 100µM Mon on A549 cells. Furthermore, Mon markedly suppressed tumor growth, decreased N-cadherin, VEGFA, p-AKT, and p-NF-κB expressions, increased apoptosis and E-cadherin expression, and SC79 reversed the inhibition effects of Mon in vivo. Thus, Mon is a potential antitumor natural drug, and more signaling pathways involved in Mon-modulated lung cancer progression are worth testing for further investigation.

The key role of the NUDT3 gene in lung adenocarcinoma progression and its association with the manganese ion metabolism family.

Lung adenocarcinoma is one of the most common malignant tumors worldwide. Its complex molecular mechanisms and high tumor heterogeneity pose significant challenges for clinical treatment. The manganese ion metabolism family plays a crucial role in various biological processes, and the abnormal expression of the NUDT3 gene in multiple cancers has drawn considerable attention. This study aims to systematically analyze the expression characteristics of the NUDT3 gene in lung adenocarcinoma and its association with tumor progression, integrating single-cell transcriptomic analysis and experimental validation using cell lines. This study employed a comprehensive set of analytical approaches. Single-cell transcriptomic analysis of two normal and four lung adenocarcinoma samples from the GSE149655 dataset was performed using the Seurat package to identify and annotate distinct cell populations, focusing on epithelial and macrophage subtypes. Non-negative matrix factorization (NMF) and gene set variation analysis (GSVA) were applied to assess the functional enrichment and expression profiles of the manganese ion metabolism family across 14 cancers. Quantitative PCR (qPCR) was conducted to evaluate the relative mRNA expression of NUDT3 in A549 lung adenocarcinoma cell lines compared to BEAS-2B normal bronchial epithelial cell lines. GAPDH was used as the reference gene for normalization, and the relative expression levels of NUDT3 in these cell lines were analyzed to confirm bioinformatics findings. NUDT3 was found to be significantly upregulated in lung adenocarcinoma and highly correlated with tumor mutation burden (TMB) and mutation enrichment (MEs). Single-cell transcriptomic analyses demonstrated elevated NUDT3 expression in lung adenocarcinoma epithelial cells, with expression levels increasing in cells associated with advanced tumor stages. Furthermore, qPCR analysis confirmed the upregulation of NUDT3 in A549 cells compared to BEAS-2B cells, consistent with transcriptomic data. These results also highlighted significant expression differences of the manganese ion metabolism family across various cancers, including BLCA, BRCA, and COAD, with notable NUDT3 mutations enriched in these cancers. This study underscores the critical role of NUDT3 in lung adenocarcinoma progression and its potential as a therapeutic target. These findings contribute to the understanding of the manganese ion metabolism family in cancer biology and provide a foundation for precision therapies targeting NUDT3 in lung adenocarcinoma.

Lidocaine Inhibits the Proliferation of Non-Small Cell Lung Cancer and Exerts Anti-Inflammatory Effects Through the TLR-9/MyD88/NF-κB Pathway.

Lung cancer represents a significant global health burden, with non-small cell lung cancer (NSCLC) being the most common subtype. The current standard of care for NSCLC has limited efficacy, highlighting the necessity for innovative treatment options. Lidocaine, traditionally recognized as a local anesthetic, has emerged as a compound with potential antitumor and anti-inflammatory capabilities. This study was designed to explore the impact of lidocaine on NSCLC cell proliferation and inflammation, particularly focusing on the Toll-like receptor 9 (TLR)-9/MyD88/NF-κB signaling pathway. A nude mice model of NSCLC was employed, with animals receiving lidocaine at different concentrations. In vitro experiments on A549 cells involved exposure to lidocaine, followed by assessment of cell viability, cytokine expression, and TLR-9 levels using the 3-(4,5-Dimethylthiazol-2-Yl)-2,5-Diphenyltetrazolium Bromide (MTT) assay, enzyme-linked immunosorbent assay, and Quantitative Real-time polymerase chain reaction (qPCR). Protein levels were evaluated via Western blot analysis. Additionally, A549 cells were transfected with a TLR-9-overexpressing lentivirus to dissect the role of TLR-9 in lidocaine's mechanism of action. Treatment with lidocaine led to a significant reduction in tumor dimensions and a decrease in inflammatory marker expression in the NSCLC mouse model. In cellular assays, lidocaine effectively suppressed A549 cell proliferation and the expression of inflammatory cytokines. The overexpression of TLR-9 partially negated the suppressive effects of lidocaine, underscoring the significance of the TLR-9/MyD88/NF-κB pathway in mediating lidocaine's effects. Lidocaine's inhibitory effects on NSCLC cell proliferation and its anti-inflammatory mechanisms are mediated through the TLR-9/MyD88/NF-κB pathway. The study's results offer promising insights into the therapeutic potential of lidocaine in NSCLC and pave the way for future investigations into its application in cancer therapy.

Chemical Profiling, Antioxidant, and Antiproliferative Activities of Algerian Cistus creticus L. Leaf Extracts: Evidence from In vitro and In Silico Studies.

Research into oxidative stress, cancer, and natural products revealed promising avenues for therapeutic intervention. Natural products are considered potent pharmaceuticals in combating oxidative stress and its relationship with cancer. This study was carried out to evaluate the chemical profile and antioxidant activities using DPPH, ABTS, Phenanthroline, Cupric, Phosphomolybdenum, FRAP, Hydroxyl, Iron chelation in vitro assays, and anticancer properties by MTT method of Cistus creticus extracts. The chemical composition was determined using the LC/MS-MS technique. Therefore, in silico methods, particularly molecular docking and dynamic simulation were applied for molecular interaction analysis. The obtained results revealed a wide variety of phenolic compounds in all studied fractions, in their qualitative and quantitative distribution. In most antioxidant assays, the butanol and ethyl acetate extracts exhibited the most effective effects, followed by the aqueous extract, while the petroleum ether and chloroform fractions exhibited much lower activity in comparison with standards. In parallel, ethyl acetate, n-butanol, and chloroform extracts exhibited potent antiproliferative activity against T47D and A549 cell lines, while the aqueous extract showed an IC50 in the range of mg/ml. Moreover, the analysis of interactions identified compounds against particular targets in studied cell lines using molecular docking showed a great affinity, especially for the ligands Hesperidin, Luteolin-7-O-glucoside and Rutin. Also, the molecular dynamic simulation of the interacting complexes Hesperidin-mTOR, Lutin-EGFR and Apigenin-HER2 revealed precise interaction, providing insights into their stability and dynamic behavior. Furthermore, the studied ligand exhibited interesting pharmacokinetic properties with no reported toxicity. These findings confirmed the potential of Algerian Cistus creticus L. leaf extracts as promising therapeutic molecules for combating oxidative stress and cancer.

Chitosan Nanoparticle-Encapsulated Cordyceps militaris Grown on Germinated Rhynchosia nulubilis Reduces Type II Alveolar Epithelial Cell Apoptosis in PM2.5-Induced Lung Injury

Chitosan nanoparticles (CNPs) were synthesized in this study to enhance the limited bioactivity and stability of Cordyceps militaris grown on germinated Rhynchosia nulubilis (GRC) and effectively deliver it to target tissues. Under optimized conditions, stable encapsulation of GRC was achieved by setting the chitosan (CHI)-to-tripolyphosphate (TPP) ratio to 4:1 and adjusting the pH of TPP to 2, resulting in a zeta potential of +22.77 mV, which indicated excellent stability. As the concentration of GRC increased, the encapsulation efficiency decreased, whereas the loading efficiency increased. Fourier-transform infrared (FT-IR) spectroscopy revealed shifts in the amide I and II bands of CHI from 1659 and 1578 to 1639 cm⁻1, indicating hydrogen bonding and successful encapsulation of GRC encapsulated with CNPs (GCN). X-ray diffraction (XRD) examination revealed the transition of the nanoparticles from a crystalline to an amorphous state, further confirming successful encapsulation. In vivo experiments demonstrated that GCN treatment significantly reduced lung injury scores in fine particulate matter (PM2.5)-exposed mice (p < 0.05) and alleviated lung epithelial barrier damage by restoring the decreased expression of occludin protein (p < 0.05). In addition, GCN decreased the PM2.5-induced upregulation of MMP-9 and COL1A1 mRNA expression levels, preventing extracellular matrix (ECM) degradation and collagen accumulation (p < 0.05). GCN exhibited antioxidant effects by reducing the mRNA expression of nitric oxide synthase (iNOS) and enhancing both the protein and mRNA expression of superoxide dismutase (SOD-1) caused by PM2.5, thereby alleviating oxidative stress (p < 0.05). In A549 cells, GCN significantly reduced PM2.5-induced reactive oxygen species (ROS) production compared with GRC (p < 0.05), with enhanced intracellular uptake confirmed using fluorescence microscopy (p < 0.05). In conclusion, GCN effectively alleviated PM2.5-induced lung damage by attenuating oxidative stress, suppressing apoptosis, and preserving the lung epithelial barrier integrity. These results emphasize its potential as a therapeutic candidate for preventing and treating the lung diseases associated with PM2.5 exposure.

Label-Free and Sequence-Independent Isothermal Amplification Strategy for the Simultaneous Detection of Genomic 5-Methylcytosine and 5-Hydroxymethylcytosine.

5-Methylcytosine (5mC) and 5-hydroxymethylcytosine (5hmC) are crucial epigenetic modifications in eukaryotic genomic DNA that regulate gene expression and are associated with the occurrence of various cancers. Here, we combined bisulfite conversion with 4-acetamido-2,2,6,6-tetramethyl-1-oxopiperridinium tetrafluoroborate (ACT+BF4-, TCI) oxidation to develop a label-free and sequence-independent isothermal amplification (BTIA) assay for a genome-wide 5mC and 5hmC analysis. The BTIA strategy can distinguish 5mC and 5hmC signatures from other bases with high sensitivity and good specificity, avoiding sophisticated chemical modifications and expensive protein labeling. Moreover, the utilization of terminal deoxynucleotidyl transferase (TdT) enables the proposed strategy to detect global 5mC and 5hmC without sequence dependence. With only 78 ng of input of genomic DNA, global 5mC and 5hmC levels were accurately quantified in cells (including cancer cells of A549, T47D, and K562 and normal cells of HEK-293T, CHO, and NRK-52E) and clinical whole blood samples (including healthy control, precancerous cervical cancer, and confirmed cervical cancer) within 18 h. The detection results suggested that 5mC was highly expressed in cancer cells. More importantly, a significant increase in 5mC was observed in precancerous cervical cancer and further upregulation in confirmed cervical cancer, suggesting a correlation between 5mC and cancer occurrence and development. However, 5hmC showed the reverse result in these tested cells and clinical samples. Collectively, the BTIA strategy can be easily performed on the ordinary heating apparatus in almost all research and medical laboratories, showing a significant application in the early screening of cervical cancer in the clinic.

Preparation of pH-Responsive Oil-in-Water Microemulsion Using a Full-Bio-Based Surfactant Containing a Dynamic Covalent Bond.

A novel pH-responsive full-bio-based surfactant (Ca-S) containing a dynamic covalent bond is synthesized using renewable cashew phenol, 5-chloro-2-furanaldehyde, and taurine. The structure of Ca-S is characterized by Fourier transform infrared spectroscopy (FTIR) and 1H nuclear magnetic resonance (NMR) analysis. Limonene containing oil-in-water (O/W) microemulsions are prepared on the basis of the Ca-S surfactant and are applied to the remediation of oil-contaminated soil under low-energy conditions at ambient temperature. These results show that Ca-S presents lower cytotoxicity against the A549 cell line. The microemulsions feature low interfacial tension and fast response characteristics, along with an optimal pH responsiveness. The microemulsions that undergo rapid and complete phase separation separate into distinct oil and water phases within 5 s in acidic media (pH < 4). In addition, the microemulsion significantly reduces the oil content to below 4%. The microemulsions hold broad application prospects in soil remediation.

Utilizing TP53 hotspot mutations as effective predictors of gemcitabine treatment outcome in non-small-cell lung cancer

TP53 mutations are recognized to correlate with a worse prognosis in individuals with non-small cell lung cancer (NSCLC). There exists an immediate necessity to pinpoint selective treatment for patients carrying TP53 mutations. Potential drugs were identified by comparing drug sensitivity differences, represented by the half-maximal inhibitory concentration (IC50), between TP53 mutant and wild-type NSCLC cell lines using database analysis. In addition, clinical data from NSCLC patients were collected to evaluate both their TP53 status and their response to gemcitabine, thereby facilitating further validation. Subsequently, NSCLC cell lines with different TP53 status (A549 and H1299) were subjected to gemcitabine treatment to investigate the association between TP53 mutations and gemcitabine response. According to the dataset, NSCLC cell lines carrying TP53 mutations displayed heightened sensitivity to gemcitabine. From a clinical standpoint, patients exhibiting TP53 hotspot mutations demonstrated prolonged overall survival upon gemcitabine treatment. In vitro, overexpressing various hotspot TP53 mutations significantly sensitized H1299 cells to gemcitabine. Moreover, the knockdown of TP53 in A549 cells notably augmented sensitivity to gemcitabine treatment, as evidenced by cell viability and reproductive cell death assays. Conversely, the overexpression of wild-type TP53 in H1299 cells led to an increased resistance against gemcitabine. Gemcitabine is a treatment option for patients with non-small cell lung cancer (NSCLC) who carry TP53 hotspot mutations. This potential effectiveness might arise from its ability to disrupt DNA damage repair processes, leading to G2/M phase cell cycle arrest or an augmentation of mitotic abnormalities, eventually cause cell death. As a result, when planning treatment strategies for NSCLC patients possessing TP53 hotspot mutations, gemcitabine should be considered to incorporate into the indication.

Biocompatible Lyotropic Nanocarriers for Improved Delivery of Ascorbyl Tetraisopalmitate in Skincare.

Ascorbyl tetraisopalmitate (VC-IP) is a novel form of ascorbic acid characterized by reduced water solubility due to complete acylation with palmitate. This study investigated the potential cosmetic application of VC-IP when encapsulated in lyotropic liquid crystal nanoparticles (VC-IP LCNPs) by using a high-pressure homogenization (HPH) method. The particle size, zeta potential, and polydispersity index (PDI) of the obtained VC-IP LCNPs were determined as 158.8 ± 3.2 nm, -35.1 ± 2.8 mV, and 0.12 ± 0.02, respectively. The drug loading (DL%) of the VC-IP LCNPs was approximately 35.1%. Morphological changes in LCNPs, transitioning from a sponge phase to vesicles, confirmed the successful loading of VC-IP, as demonstrated by transmission electron microscopy (Cryo-TEM) and small-angle X-ray scattering (SAXS) experiments. The sustained release of VC-IP was also observed through the Franz transdermal diffusion test, indicating that VC-IP LCNPs facilitated the sustained-release effect of VC-IP into the skin. VC-IP LCNPs exhibited good biocompatibility, showing nontoxicity to HaCaT cells and zebrafish embryos. Raman distribution imaging confirmed that VC-IP successfully penetrated the stratum corneum and reached the dermis. In assessments of whitening effects, VC-IP LCNPs significantly the reduced reactive oxygen species (ROS) content in zebrafish and melanin areas in the heads of zebrafish. Furthermore, VC-IP LCNPs effectively inhibited tyrosinase activity and the proliferation of A375 cells compared with pure VC-IP. Additionally, VC-IP LCNPs significantly reduced the melanin area in the heads of zebrafish. Therefore, the developed VC-IP LCNPs present a promising carrier for the enhanced application of active ingredients, such as VC-IP in whitening products.

Intervention of a Communication Between PI3K/Akt and β-Catenin by (-)-Epigallocatechin-3-Gallate Suppresses TGF-β1-Promoted Epithelial-Mesenchymal Transition and Invasive Phenotype of NSCLC Cells.

The epithelial-mesenchymal transition (EMT) assists in the acquisition of invasiveness, relapse, and resistance in non-small cell lung cancer (NSCLC) and can be caused by the signaling of transforming growth factor-β1 (TGF-β1) through Smad-mediated or Smad-independent pathways. (-)-Epigallocatechin-3-gallate (EGCG), a multifunctional cancer-preventing bioconstituent found in tea polyphenols, has been shown to repress TGF-β1-triggered EMT in the human NSCLC A549 cell line by inhibiting the activation of Smad2 and Erk1/2 or reducing the acetylation of Smad2 and Smad3. However, its impact on the Smad-independent pathway remains unclear. Here, we found that EGCG, similar to LY294002 (a specific inhibitor of phosphatidylinositol 3-kinase [PI3K]), downregulated Akt activation and restored the action of glycogen synthase kinase-3β (GSK-3β), accompanied by TGF-β1-caused changes in hallmarks of EMT such as N-cadherin, E-cadherin, vimentin, and Snail in A549 cells. EGCG inhibited β-catenin expression and its nuclear localization caused by TGF-β1, suggesting that EGCG blocks the crosstalk between the PI3K/Akt/GSK-3β route and β-catenin. Furthermore, it was shown that EGCG suppressed TGF-β1-elicited invasive phenotypes of A549 cells, including invading and migrating activities, matrix metalloproteinase-2 (MMP-2) secretion, cell adhesion, and wound healing. In summary, we suggest that EGCG inhibits the induction of EMT by TGF-β1 in NSCLC not only through a Smad-dependent pathway, but also through the regulation of the PI3K/Akt/β-catenin signaling axis.

Substituent Effect on Cytotoxicity and Interaction Profile of DNA/BSA With Selected Terpyridine Derivatives of Ester/Ferrocene and Their Fe(III) and Ru(III) Complexes—A Comparative Analysis

ABSTRACTThe present work demonstrates the synthesis of ester and ferrocene groups containing new terpyridine‐based ligands (FRL and RL) and their selected meal complexes FR‐Fe, FR‐Ru, RL‐Fe, and RL‐Ru. Investigations on the influence of the ester and ferrocene groups on binding to DNA/BSA and anticancer activity were carried out. All these products were structurally characterized by using spectroscopic and analytical techniques. Notably, the molecular structures of both the ligands and the ester‐based Fe(III) complex were confirmed by single‐crystal X‐ray diffraction analysis. Additionally, theoretical calculations were employed to further establish the structures of the synthesized ligands and their metal complexes, which were subsequently utilized for molecular docking studies to evaluate their binding potential. The docking results revealed the best binding orientation with the lowest possible binding energy of −10.20 kcal/mol for the RL‐Fe complex with BSA and −8.30 kcal/mol for the FR‐Fe complex with DNA. Furthermore, the binding interactions of all the synthesized compounds were investigated using UV–visible absorption (UV) and fluorescence spectroscopy (FL). The binding sites and binding constants were determined using the Stern‐Volmer equation. Moreover, cytotoxicity assays were performed against lung cancer (A549), liver cancer (Hep G2), and normal Vero cell lines, with results benchmarked against cisplatin. Notably, the ligand (FRL) exhibited significant cytotoxic action with an IC50 of 54.51 μM compared to the related Fe(III) complex (FR‐Fe) (i.e., 69.86 μM) against A549 cells. But the complex FR‐Fe displayed IC50 of 56.19 μM against Hep G2 cells, demonstrating better potency over cisplatin.

Antioxidant Peptides from Hizikia fusiformis: A Study of the Preparation, Identification, Molecular Docking, and Cytoprotective Function of H2O2-Damaged A549 Cells by Regulating the Keap1/Nrf2 Pathway

Hijiki (Hizikia fusiformis) is a seaweed native to warm-temperate and subtropical regions that has a high edible value and economic value, with a production of about 2 × 105 tons/year. Current research has clearly shown that the pharmacological activities of active ingredients from hijiki have covered a broad spectrum of areas, including antioxidant, hypoglycemic, antiviral, anticoagulant, anti-inflammatory, intestinal flora modulation, anti-aging, antineoplastic and antibacterial, and anti-Alzheimer’s disease areas. However, no studies have reported on the production of antioxidant peptides from hijiki proteins. The objectives of this study were to optimize the preparation process and explore the cytoprotective function and mechanisms of antioxidant peptides from hijiki protein. The results indicated that papain is more suitable for hydrolyzing hijiki protein than pepsin, trypsin, alkaline protease, and neutral protease. Under the optimized parameters of an enzyme dosage of 3%, a material–liquid ratio of 1:30, and an enzyme digestion time of 5 h, hijiki hydrolysate with a high radical scavenging activity was generated. Using ultrafiltration and serial chromatographic methods, ten antioxidant oligopeptides were purified from the papain-prepared hydrolysate and identified as DGPD, TIPEE, TYRPG, YTPAP, MPW, YPSKPT, YGALT, YTLLQ, FGYGP, and FGYPA with molecular weights of 402.35, 587.61, 592.64, 547.60, 532.53, 691.77, 523.57, 636.73, 539.58, and 553.60 Da, respectively. Among them, tripeptide MPW could regulate the Keap1/Nrf2 pathway to significantly ameliorate H2O2-induced oxidative damage of A549 cells by increasing cell viability and antioxidant enzyme (SOD, CAT, and GSH-Px) activity, decreasing ROS and MDA levels, and reducing the apoptosis rate. Molecular docking experiments show that HFP5 (MPW) exerts its inhibitory effect mainly through hydrogen bonds and hydrophobic interactions with the Kelch domain of the Keap1 protein, eventually facilitating the translocation of Nrf2 to the nucleus. Therefore, antioxidant peptides from hijiki can be applied to develop algae-derived health foods for treating diseases associated with oxidative stress.

Antiviral effects and mechanism of Qi pi pill against influenza viruses.

Qi pi pill (QPP), which contains Renshen, Baizhu, Fuling, Gancao, Chenpi, Shanyao, Lianzi, Shanzha, Liushenqu, Maiya, and Zexie, was recommended for preventing and treating COVID-19 in Shandong Province (China). However, the mechanism by which QPP treats infectious diseases remains unclear. This study aims to investigate the therapeutic effect of QPP invitro and on acute influenza infection in mice, exploring its mechanism of action against influenza A virus (IAV). The invitro activity of QPP was assessed using dose-response curve analysis and titer reduction assay, and its antiviral mechanism was identified invitro by real-time polymerase chain reaction (PCR), time-of-addition, and enzymatic assays. The antiviral efficacy of QPP was further evaluated invivo using BALB/c mice infected with IAV. At the same time, each single Chinese herbal medicine in QPP was evaluated to preliminarily identify those with antiviral effects. In vitro results showed that QPP exhibited a higher potency antiviral effect against both influenza A and B viruses, inhibiting viral RNA replication and release by targeting RNA-dependent RNA polymerase and neuraminidase. Additionally, QPP significantly decreased the expression of inflammatory cytokines in A549 cells. Invivo study revealed that QPP significantly reduced the lung index and viral load in lung tissue of mice infected with IAV. Renshen, Gancao, Zexie, and Lianzi were the Chinese herbal medicines from QPP that showed anti-IAV activity. The antiviral activity of QPP targets IAV replication and release, cytokine modulation in host cells, and provides protection in mice with acute influenza infection.

Pyrogallol acts as a novel anticancer factor to enhance the sensitivity to cisplatin in ovarian cancer cells through inducing miR-15a upregulation

IntroductionPyrogallol is a catechin compound, as an active ingredient extracted from Emblica officinalis, has antiproliferative and antiapoptotic effects on some of the human cancer cells. Cisplatin is known as an effective chemotherapeutic agent against cancer by inducing DNA damage and apoptosis. However, the effect of pyrogallol alone and in combination with cisplatin on proliferation and apoptosis of ovarian cancer has not yet been reported.MethodsIn the present study, ovarian carcinoma cell line A2780 was cultivated, and the anticancer properties of pyrogallol and cisplatin determined by cell treatment of the two drugs separately and combination. Cell viability and apoptosis rate was investigated by MTT assay and flow cytometry. The effects of the compounds in inhibiting colony formation and migration were measured by colony formation and scratch methods.ResultsThe results of the present study revealed that pyrogallol significantly inhibited the proliferation of A2780 cells in a dose-dependent manner. The combination treatment of pyrogallol and cisplatin exhibited a prominent inhibitory effect on the growth, colony formation and migration, and increased the induction of apoptosis up to 2.5 times compared to single treatments. Also, the qPCR results showed a significant increase in the expression of miR-15a and a decrease in the expression of miRNA target genes under treatment with the drugs both separately and in their combination.ConclusionThis study suggests that the combination treatment of pyrogallol and cisplatin significantly increases the death of ovarian cancer cells by inducing apoptosis. Therefore, it can boost the efficacy of chemotherapy and reduce its side effects. It is hoped that the findings of the present study can provide a new perspective for pyrogallol as a therapeutic agent in the treatment of ovarian cancer.

Synergistic Anti-Cancer Activities of Curcumin Derivative CU17 Combined with Gemcitabine Against A549 Non-Small-Cell Lung Cancer Cells

Recently, the curcumin derivative CU17 possessing HDAC inhibitory activity has been shown to synergistically enhance the anti-proliferative activity of Gem against lung cancer cells. Nevertheless, the mechanism(s) underlying the synergistic anti-cancer effect remains to be investigated. This study aimed to investigate the mechanisms that underpin the anti-cancer activity of the combined Gem and CU17 against NSCLC A549 cells both in vitro and in mouse xenograft models. CU17 was successfully synthesized and subsequently investigated for its combination effects with Gem on inductions of cell cycle arrest and apoptosis in A549 cells. The combination treatment substantially decreased cell survival through S phase prolongation and G2/M phase cell cycle arrest via up-regulating the expressions of p21 and p53 proteins. Additionally, CU17 potentiated the apoptotic effect of Gem in A549 cells by increasing the Bax/Bcl-2 ratio. The co-treatment resulted in an up-regulation of pERK1/2 and Ac-H3 expression. An in vivo study demonstrated that CU17 significantly improved the anti-cancer effect of Gem in nude mice utilizing A549 cell xenografts. The hematoxylin and eosin (H&amp;E) staining results indicated that CU17 decreased the toxicity of Gem to the liver, kidneys, and spleen. Overall, CU17 enhanced the effectiveness of Gem while decreasing its toxicity. This compound shows promise as a chemosensitizer for NSCLC treatment with Gem.

Upregulation of phosphatase and tensin homolog deleted on chromosome ten inhibits lung cancer cell proliferation by suppressing the oncogene polo-like kinase 1 and inducing autophagy

Objective: Lung cancer is one of the main causes of cancer-related mortality globally, and it poses considerable therapeutic challenges. Polo-like kinase 1 (PLK1) exhibits upregulation in lung cancer, and PLK1 silencing promotes autophagy in lung cancer cells, which inhibits tumor progression. The phosphatase and tensin homolog deleted on chromosome ten (PTEN) acts as a tumor suppressor gene. This study aimed to investigate whether PTEN regulates autophagy and inhibits lung cancer-cell proliferation by suppressing PLK1. Material and Methods: In this study, we evaluated cell proliferation by silencing or overexpressing PLK1 and PTEN in A549 cells through 5-ethynyl-2'-deoxyuridine labeling and cloning experiments. The autophagy levels were detected through transmission electron microscopy, real-time quantitative polymerase chain reaction, and Western blot. Finally, the results of in vitro experiment were further verified using an in vivo xenograft tumor animal model. Results: The upregulation of PTEN suppressed PLK1 expression in lung cancer cells and reduced their proliferation rate. In addition, the overexpression of PTEN has been associated with the growth of lung cancer tumors. In parallel, the levels of autophagy of lung cancer cells rose in response to PTEN upregulation in vivo and in vitro. Conclusion: This study revealed that PTEN promotes the autophagy of lung cancer cells and inhibits cell proliferation and tumor growth by suppressing PLK1 expression. This finding provides a new strategy for lung cancer treatment by utilizing the autophagy-regulating effect of PTEN to inhibit lung cancer growth by targeting PLK1.

Exploring miR-21 Knock-Out Using CRISPR/Cas as a Treatment for Lung Cancer

Background: Lung cancer is a leading cause of cancer-related deaths worldwide. Its high incidence and poor prognosis demonstrate the need to investigate new therapies. The PI3K/AKT pathway is activated in carcinogenic processes such as invasion, proliferation, and drug resistance. MiR-21 is a microRNA overexpressed in numerous types of cancer and which activates PI3K/AKT pathway by down-regulating its main targets, PTEN and PDCD4. CRISPR is a revolutionary gene-editing technology that allows genes to be deleted. The aim of this study was to use CRISPR/Cas9 technology as an option to reduce carcinogenic and drug resistance processes by eliminating miR-21. Methods: CRISPR/Cas9 was used to knock out miR-21 (miR-21 KO) in A549 lung cancer cells and thus reverse the carcinogenic processes activated by miR-21 overexpression. Furthermore, the effect of miR-21 KO on drug resistance was studied, choosing the main chemotherapeutic agents used for the treatment of lung cancer: gemcitabine, carboplatin, paclitaxel, and oxaliplatin. Results: miR-21 KO A549 cells exhibited a reduction in proliferation, migration, and colony formation compared to A549 cells. In contrast, the expression of PTEN and PDCD4 increased in miR-21 KO A549 cells. Furthermore, miR-21 KO A549 cells showed a decrease in the IC50 of the drugs used for the treatment of lung cancer: gemcitabine, carboplatin, paclitaxel, and oxaliplatin. Conclusions: Based on these results, miR-21 knock-out using CRISPR/Cas could be a promising strategy for the treatment of lung cancer.

Fibrinogen Alpha Chain as a Potential Serum Biomarker for Predicting Response to Cisplatin and Gemcitabine Doublet Chemotherapy in Lung Adenocarcinoma: Integrative Transcriptome and Proteome Analyses

Cisplatin combined with gemcitabine, a doublet regimen, is the first-line treatment for patients with advanced lung adenocarcinoma (ADC); however, the treatment response remains poor. This study aimed to identify potential biomarkers for predicting response to cisplatin and gemcitabine. Tissue transcriptome and blood proteome analyses were conducted on 27 patients with lung ADC. Blood-derived proteins that reflected tissue-specific biomarkers were obtained using Venn diagrams. The candidate proteins were validated by Western blotting. Lentivirus-mediated short hairpin RNA interference was used to verify the functional roles of the candidate proteins in human A549 cells. We identified 417 differentially expressed genes, including 52 upregulated and 365 downregulated genes, and 31 differentially expressed proteins, including 26 upregulated and 5 downregulated proteins. Integrative analysis revealed the presence of alpha-1-acid glycoprotein 1 (A1AG1) and fibrinogen alpha chain (FGA or FIBA) in both the tissue and serum. FGA levels were elevated in responders compared to non-responders, and reduced serum FGA levels were correlated with resistance to this regimen. Moreover, FGA knockdown in A549 cells resulted in resistance to the doublet regimen. Our findings indicate that FGA is a tissue-specific serum protein that may function as a blood-based biomarker to predict the response of patients with lung ADC to cisplatin plus gemcitabine chemotherapy.

3D Bioprinted Multidrug Resistance (MDR)-Dependent Tumor Spheroids.

Multidrug resistance (MDR) refers to the ability of cancer cells to resist various anticancer drugs and release them from the cells. This phenomenon is widely recognized as a significant barrier that must be overcome in chemotherapy. MDR varies depending on the number and expression level of the ATP-binding cassette transporter (ABC transporter), which is expressed differently in various cancer cells. Therefore, the dose of anticancer drugs should be adjusted according to the extent of MDR. The demand for drug screening that considers the differences in MDR is increasing in the process of drug discovery. In this study, three types of tumor spheroids were fabricated from HeLa (MRP1-/BCRP-), HepG2 (MRP1+/BCRP-), and A549 cells (MRP1+/BCRP+) using three-dimensional (3D) bioprinting. The fabricated tumor spheroids maintained their own MDR phenotypes. The EC50 values of doxorubicin (DOX) against the three tumor spheroids were more than 2-fold higher than those against the 2D cells. In addition, the EC50 value of DOX against tumor spheroids was proportional to the number of ABC transporters. The EC50 value of DOX against A549 tumor spheroids had the largest value of 9.5 μM among the three spheroids. In addition, the EC50 values of DOX against HepG2 and A549 tumor spheroids were remarkably reduced when they were treated with ABC transporter inhibitors, such as MK-571 against MRP1 and/or NOV against BCRP. These results demonstrate the successful construction of a 3D bioprinting-based screening platform to quantitatively evaluate the anticancer efficacy of chemodrugs, considering the MDR of cancer cells.

High-Glucose-Induced Metabolic and Redox Alterations Are Distinctly Modulated by Various Antidiabetic Agents and Interventions Against FABP5/7, MITF and ANGPTL4 in Melanoma A375 Cells

Hyperglycemia-induced effects on cellular metabolic properties and reactive oxygen species (ROS) generation play pivotal roles in the pathogenesis of malignant melanoma (MM). This study assessed how metabolic states, ROS production, and related gene expression are modulated by antidiabetic agents. The anti-diabetic agents metformin (Met) and imeglimin (Ime), inhibitors of fatty acid-binding proteins 5/7 (MF6) and microphthalmia-associated transcription factor (MITF) (ML329), and siRNA-mediated knockdown of angiopoietin-like protein 4 (ANGPTL4), which affect mitochondrial respiration, ROS production, and related gene expression, were tested in A375 (MM cell line) cells cultured in low (5.5 mM) and high glucose (50 mM) conditions. Cellular metabolic functions were significantly and differently modulated by Met, Ime, MF6, or ML329 and knockdown of ANGPTL4. High glucose significantly enhanced ROS production, which was alleviated by Ime but not by Met. Both MF6 and ML329 reduced ROS levels under both low and high glucose conditions. Knockdown of ANGPTL4 enhanced the change in glucose-dependent ROS production. Gene expression related to mitochondrial respiration and the pathogenesis of MM was significantly modulated by different glucose conditions, antidiabetic agents, MF6, and ML329. These findings suggest that glucose-dependent changes in cellular metabolism and redox status are differently modulated by antidiabetic agents, inhibition of fatty acid-binding proteins or MITF, and ANGPTL4 knockdown in A375 cells.