Human Lines Research Articles

New Pd(II)-pincer type complexes as potential antitumor drugs: synthesis, nucleophilic substitution reactions, DNA/HSA interaction, molecular docking study and cytotoxic activity.

Two new complexes of Pd(II), [Pd(L1)Cl]Cl (Pd1) and [Pd(L2)Cl]Cl (Pd2), (where L1 = N2,N6-bis(5-methylthiazol-2-yl)pyridine-2,6-dicarboxamide and L2 = N2,N6-di(benzo[d]thiazol-2-yl)pyridine-2.6-dicarboxamide) were synthesized. Characterization of the complexes was performed using elemental analysis, IR, 1H NMR spectroscopy and MALDI-TOF mass spectrometry. The nucleophilic substitution reactions of complexes with L-Methionine (L-Met), L-Cysteine (L-Cys) and guanosine-5'-monophosphate (5'-GMP) were studied by stopped-flow method at physiological conditions (pH = 7.2 and 37 °C). Complex Pd1 was more reactive than Pd2 in all studied reactions, while the order of reactivity of the selected ligands was: L-Met > L-Cys > 5'-GMP. The interaction of complexes with calf thymus-DNA (CT-DNA) was studied by Uv-Vis absorption and fluorescence emission spectroscopy. Competitive binding studies with intercalative agent ethidium bromide (EB) and minor groove binder Hoechst 33258 were performed as well. Both complexes interacted with DNA through intercalation and minor groove binding, where the latter was preferred. Additionally, the interaction of Pd1 and Pd2 complexes with human serum albumin (HSA) was studied employing fluorescence quenching spectroscopy. The results indicate a moderate binding affinity of complexes, with slightly stronger binding of the Pd1. Fluorescence competition experiments with site-markers (eosin Y and ibuprofen) for HSA were used to locate the binding site of Pd1 to the HSA. Additionally, the interaction with DNA and HSA was studied by molecular docking and the revealed results were in good agreement with the experimentally obtained ones. Pd1 complex exhibited cytotoxicity toward human (HCT116) and mouse cell lines (CT26) of colorectal cancer, mouse (4T1) and human (MDA-MB468) breast cancer lines and non-cancerous mouse mesenchymal stem cells (mMSC). In addition, Pd1 complex demonstrated significant selectivity towards cancer cells over non-cancerous mMSC, indicating a high potential to eliminate malignant cells without affecting normal cells. It induced apoptosis in CT26 cells, effectively arrested the cell cycle in the S phase, and selectively down-regulated cyclin D and cyclin E. Moreover, it can alter the expression of cell cycle regulators by increasing p21 and decreasing p-AKT. These findings confirm its ability to disrupt key tumor cell survival signals and suggest that the Pd1 complex is a potent candidate for effective cancer treatment.

Circular Makerspaces as Alternative Employment Platforms for Circular Jobs

AbstractTransitioning towards a circular economy requires holistic consideration encompassing environmental, economic, and social dimensions. This perspective paper explores circular makerspaces as innovative platforms for fostering social integration and creating employment opportunities within the circular economy, as makerspaces can offer a more inclusive alternative to traditional employment platforms. They have the potential to unveil unrecognised talents, bridge access to under-utilised human capital, and act as pivotal conduits to a decent and inclusive circular labour force. Drawing insights from the European Horizon 2020 project: Pop-Machina, this perspective paper emphasizes the importance of collaborative efforts among policymakers, practitioners, and researchers to unlock the full transformative potential of circular makerspaces. By prioritizing the social aspect of sustainability and leveraging the network of circular makerspaces, circular makerspaces can unlock unexplored human capital, provide employment opportunities and cultivate inclusive, sustainable communities, while highlighting their potential for societal empowerment and innovation. Finally, this perspective paper underscores the need for ongoing research and collaboration to comprehensively understand and evaluate the role of circular makerspaces in the inclusive circular labour market, ensuring that the social dimension remains central to sustainable development endeavours and informing effective policy making.This project focused on the gene PFKFB3, a pivotal regulator of the Warburg effect, which facilitates enhanced glycolysis in cancer cells, including MCF7 human breast cancer lines. The objective was to investigate the effects of PFKFB3 knockout on the metabolic profile and proliferative capacity of MCF7 cells, hypothesizing that disruption of this gene would significantly impair cellular energy production and, consequently, cell growth. Through the application of CRISPR/Cas9 technology, PFKFB3 was specifically targeted and knocked out, followed by a meticulous process of selection to enrich for cells bearing the knockout. Growth assays, particularly MTT, were conducted to evaluate the impact of PFKFB3 deletion on cell proliferation. Contrary to expectations preliminary results indicated no significant difference in the growth rates between PFKFB3-knockout and control groups. This outcome suggests possible metabolic flexibility within MCF7 cells, allowing them to bypass the blockade of one glycolytic pathway—a concept supported by current understandings of cancer metabolism. Despite the lack of expected growth inhibition, this study provides critical insights into the adaptability of cancer cells to metabolic interventions and highlights the importance of targeting multiple metabolic pathways. Future directions will consider exploring the combined knockout of PFKFB3 and other glycolytic genes to overcome the metabolic resilience of cancer cells, thereby offering a more effective strategy for crippling cancer cell energy supplies.

Phenolic Contents, Antioxidant Activities, LCMS Profiles of Mespilus germanica Leaf Extract and Effects on mRNA Transcription Levels of Apoptotic, Autophagic, and Necrotic Genes in MCF7 and A549 Cancer Cell Lines.

Cancer, defined by the continuous, uncontrollable proliferation of cells in the human body, is a disease with a rapidly increasing incidence and mortality rate. Scientists are looking for novel ways to cure and prevent this sneaky disease because of the toxicity of contemporary chemotherapy and the cancer cells' resilience to anticancer drugs. Determining the effect of herbal medicines, which do not have as harmful side effects as synthetic drugs, on cancer cell lines is an essential preliminary study in the production of effective drugs against cancer. In this study, the phenolic acid profile, antioxidant capacity, and cytotoxicity of the medicinal plant Mespilus germanica (MG) leaf extract were determined, and its effects on the expression of some apoptotic, necrotic, and autophagic pathway genes of MCF7 (Human breast cancer line) and A549 (Human lung cancer line) and healthy HDF (Human Dermal Fibroblasts) cells were investigated for the first time. The LCMS device detected many important phenolic compounds previously reported to act against cancer cells in Mespilus germanica leaf extract. DPPH and total phenolic content showed high antioxidant capacity. The cytotoxicity of MG was determined by the MTT method. The levels of mRNA transcription for Atg5, Atg3, Rıpk1, Bcl2, Bax, Apaf1, Caspase-8, Caspase-7, Caspase-3, and Caspase-9, as well as the expression patterns of the DNA damage markers P53 and Parp-1 genes, were assessed. MG leaf extract did not cause significant toxicity against healthy HDF cells. However, it had a cytotoxic effect on A549 and MCF7 cancer cell lines, increasing the transcription levels of essential genes involved in cell death mechanisms. This research is the first to analyze the phenolic components and antioxidant capabilities of leaf extracts from Mespilus germanica. Additionally, it investigates the impact of these extracts on crucial genes involved in cell death pathways of A549 lung cancer, MCF7 breast cancer, and non-cancerous HDF (Human Dermal Fibroblasts) cells.

Binding Mechanisms and Therapeutic Activity of Heterocyclic Substituted Arylazothioformamide Ligands and Their Cu(I) Coordination Complexes.

Finding new sources of biologically active compounds for anticancer or antimicrobial therapies remains an active area of research. Azothioformamides (ATFs) with a 1,3 N=N-C=S heterodiene backbone are a new class of biologically active compounds that chelate metals (e.g., Cu) forming stable ATF metal coordination complexes. In this study, ATF ligands were prepared with pyrrolidine, piperidine, N-methylpiperazine, and morpholine substituents on the formamide as to add more heterocyclic drug-like character for biological studies. Formamide derivatives were then complexed with various Cu(I) salts to form coordination complexes. Cu(I) salts were selected as to create potential bioactive compounds with less toxicity. Binding association constants of each Cu(I) salt to ATF ligands were extrapolated from UV-vis titration studies and were corroborated with DFT calculations using a hybrid functional B3LYP method. It was observed that the smaller pyrrolidine functionalized ATFs bound to the Cu(I) salts had stronger binding than any of the larger six-membered-ring heterocycles with association values in the 104 - 105 M-1 range. The ATF-Cu(I) salt coordination complexes were then evaluated for antimicrobial activity against two bacteria (Staphylococcus aureus, Escherichia coli), one yeast (Candida albicans), four human cancer lines (A-549, K-562, HT-1080, MDA-MB-231), and two normal human lines (MRC-5, HFF). The ATF ligands themselves were inactive against all microbes and most human lines except K-562 cells, which were sensitive to three of the four ligands (IC50's = 7.0-25.5 μM). Most ATF-Cu(I) complexes showed low to medium micromolar activity against Candida albicans (IC50's 2.6-24.8 μM) and Staphylococcus aureus (IC50's = 3.4-37.7 μM), with increasing activity corresponding to complexes with higher binding association constants. The antiproliferative properties of ATF-Cu(I) metal salt complexes against mammalian cells were mixed, with low to medium micromolar activity across all cell lines. Notably, several ATF-Cu(I) salt coordination complexes showed submicromolar activity against the HT-1080 fibrosarcoma line (0.52-0.69 μM). The results demonstrate promising activity of ATF-Cu(I) complexes, particularly with pyrrolidine as the formamide component. These studies suggest that the stronger binding association values correlate to higher levels of biological activity.

Larvicidal, bactericidal, antioxidant, hypoglycemic effects and anti-proliferative activity of phyto-synthesized Ag-ZnO nanocomposites using Eranthemum roseum (Vahl) R.Br. leaf extract against Aedes aegypti and Culex quinquefasciatus mosquito vectors

This research is conducted using zinc doped silver nanocomposite on Eranthemum roseum leaf extract. E. roseum has its own medicinal properties due its bioactive compounds. Silver and Zinc also has its own medicinal properties due to its microbial properties. Our work is aimed to combine nanoparticles with plant extract to explore the medicinal properties. The synthesized Ag-ZnO NCs is characterized using X-ray powder diffraction (XRD), Fourier-transform infrared spectroscopy (FTIR), Scanning electron microscopy with energy-dispersive X-ray spectroscopy (SEM-EDAX) and Transmission Electron Microscopy (TEM) analysis. The XRD analysis confirms the crystal nature of the Ag-ZnO NCs. The average shape of the nanocomposite was obtained using SEM as spherical, EDAX analysis confirming the presence of Ag, Zn and Oxygen and TEM conforms the oval shape of nanoparticles Antibacterial activity shows highest level of activity against K. pneumoniae and B. subtilis. DPPH Activity of Ag-ZnO NCs highest inhibition of 52.56 % while the ABTS analysis of Ag-ZnO NCs shows 54.28 %. Hypoglycemic activity α-amylase and α- glucosidase assay of Ag-ZnO NCs obtained good activity with maximum level of inhibition 63 % and 60 %. Anti-proliferative assay shows good activity against cancer causing cells in HepG2 human liver cancer line with 67.11 %. Larvicidal potential of Ag-ZnO NCs recorded good mortality rate against Aedes aegypti and Culex quinquefasciatus. This study confirms the maximum activity of Ag-ZnO NCs and acts as Larvicidal, Anti-proliferative, Antioxidant, Hypoglycemic agents.

Retracted] NR4A1‑induced increase in the sensitivity of a human gastric cancer line to TNFα‑mediated apoptosis is associated with the inhibition of JNK/Parkin‑dependent mitophagy.

Following the publication of this paper, it was drawn to the Editor's attention by a concerned reader that certain of the data shown in Figs. 2A and 4F were strikingly similar to data appearing in different form in other articles written by different authors at different research institutes that were submitted to their respective journals at around the same time; moreover, the same data had apparently been included in the western blots featured in Fig. 5A to show the Parkin and mito‑LCIII protein bands. As it was not clear what had been the original venue for the submission of the strikingly similar data here, the Editor requested that the authors send to us all the raw data underlying the affected figures; however, the authors were not able to comply with this request at the time of asking. Given that the authors were unable to provide the supporting data as requested, the Editor of International Journal of Oncology has decided that this paper should be retracted from the Journal on account of a lack of confidence in the presented data. The authors were asked for an explanation to account for these concerns, but the Editorial Office did not receive a satisfactory reply. The Editor apologizes to the readership for any inconvenience caused. [International Journal of Oncology 52: 367‑378, 2018; DOI: 10.3892/ijo.2017.4216].

Latrophilins as Downstream Effectors of Androgen Receptors including a Splice Variant, AR-V7, Induce Prostate Cancer Progression.

Latrophilins (LPHNs), a group of the G-protein-coupled receptor to which a spider venom latrotoxin (LTX) is known to bind, remain largely uncharacterized in neoplastic diseases. In the present study, we aimed to determine the role of LPHNs in the progression of prostate cancer. We assessed the actions of LPHNs, including LPHN1, LPHN2, and LPHN3, in human prostate cancer lines via their ligand (e.g., α-LTX, FLRT3) treatment or shRNA infection, as well as in surgical specimens. In androgen receptor (AR)-positive LNCaP/C4-2/22Rv1 cells, dihydrotestosterone considerably increased the expression levels of LPHNs, while chromatin immunoprecipitation assay revealed the binding of endogenous ARs, including AR-V7, to the promoter region of each LPHN. Treatment with α-LTX or FLRT3 resulted in induction in the cell viability and migration of both AR-positive and AR-negative lines. α-LTX and FLRT3 also enhanced the expression of Bcl-2 and phosphorylated forms of JAK2 and STAT3. Meanwhile, the knockdown of each LPHN showed opposite effects on all of those mediated by ligand treatment. Immunohistochemistry in radical prostatectomy specimens further showed the significantly elevated expression of each LPHN in prostate cancer, compared with adjacent normal-appearing prostate, which was associated with a significantly higher risk of postoperative biochemical recurrence in both univariate and multivariable settings. These findings indicate that LPHNs function as downstream effectors of ARs and promote the growth of androgen-sensitive, castration-resistant, or even AR-negative prostate cancer.

Bismuth(III) triflate: an economical and environmentally friendly catalyst for the Nazarov reaction.

We describe the use of bismuth(III) triflate as an efficient and environmentally friendly catalyst for the Nazarov reaction of aryl vinyl ketones, leading to the synthesis of 3-aryl-2-ethoxycarbonyl-1-indanones and 3-aryl-1-indanones. By changing the temperature and reaction time, it was possible to modulate the reactivity, allowing the synthesis of two distinct product classes (3-aryl-2-ethoxycarbonyl-1-indanones and 3-aryl-1-indanones) in good to excellent yield. The reaction did not require additives and was insensitive to both air and moisture. Preliminary biological evaluation of some indanones showed a promising profile against some human cancer line cells.

Archaeobotanical Elucidation of <i>Moringa oleifera</i>: It’s Biological, Ethnopharmacological and Phytochemical Activities

Moringa oleifera (family Moringaceae) was originally first found in India now it has been cultivated in various regions of the world. The architecture of a plant is an endogenous morphological growth pattern through which the plant develops its shape. Analysing plant architecture is important for the understanding of plant growth, and branching patterns. A high source of natural phytochemical compounds is present in Moringa, and plant parts have high nutritional value. Moringa is used in animal feeding systems to fight malnutrition in India and Africa. The potency of Moringa is evaluated for its effectiveness on rats and humans, among other animals. It is a safe, affordable, and effective pharmacological and dietary option. M. oleifera is a fantastic food crop with clear economic, medicinal, and nutritional advantages. Moringa contains Alkaloids, vincristine, vinblastine, kaempferitrin, isoquercitrin, kaempherol, polyphenols and flavonoids that may be present in Moringa. Moringa leaves contain vitamin C and beta-carotene which act against free radicals. Quercetin acts as an antioxidant property and chlorogenic acid, polyphenol act as an antioxidant and anticancer activity. The presence of 4-L-rhamnosyloxybenzyl isothiocyanate is responsible for showing the antibacterial activity of the root. Moringa is used as a superfood supplement and has immune booster properties. Ethnopharmacological activity of Moringa includes antibacterial, anti-inflammatory, antioxidant, antidiabetic, analgesic, chronic periodontitis, antipyretic, cardio protecting, anti-asthmatic, antispasmodic, promoting breast milk production, biogas production and water filtration properties. Numerous studies have shown that it can control physiological processes, as well as both prevent and treat illnesses. Nowadays researchers developed Moringa oleifera leaf nanoparticles that show antidiabetic and antiproliferative activity against human cancer lines and others. In clinical or human studies, lower doses of Moringa were not associated with any negative effects or toxicities.

Abstract 277: Cancer-associated fibroblasts, educated by tumor cells, differentially facilitate cancer cell invasion in cell-line based human gastric tumoroids

Abstract Purposes: Gastric cancer can be categorized into two subtypes: intestinal and diffuse. The latter is notorious for frequent peritoneal metastasis, resulting in a poorer prognosis. Cancer-associated fibroblasts (CAFs) in the tumor microenvironment play crucial roles in tumor invasion and metastasis. However, little is known about whether CAFs are associated with the differential progression between the two subtypes of gastric tumors. This study investigated whether differential cancer invasion may be attributed to tumor-specific CAF activation, using a cell-line-based human gastric tumoroid model. Methods: Heterotypic tumor spheroids (TS) were prepared with human gastric cancer lines of intestinal (MKN28, MKN74) and diffuse type (MKN45, KATO III) mixed with human gastric fibroblasts (HGFs) using a minipillar-array chip previously developed. Invasion phenotype and expression of epithelial-mesenchymal transition (EMT)-related proteins were evaluated in heterotypic TSs. mRNA expression in HGFs isolated from heterotypic TSs was analyzed and compared between the two subtypes. Results: While both heterotypic TSs displayed increased invasion by HGFs, the diffuse type TS showed a higher level of invasion with individual or amoeboid migration and loose binding of cancer cells with HGF. In contrast, the intestinal type TS showed collective migration and tight binding between cancer cells and HGFs. HGFs in each heterotypic TS exhibited differential morphology, matrix remodeling patterns, and gene expression in a tumor subtype-specific manner. Compared to normal HGFs, HGFs isolated from diffuse heterotypic TS showed enriched expression of cytoskeleton-associated genes and several integrins, whereas those of intestinal heterotypic TS exhibited increased expression levels of inflammatory cytokines. Conclusions: This study established tumoroid models for human gastric cancer subtypes, intestinal and diffuse, using heterotypic TS culture. Our results suggest that the subtype-specific invasion ability of tumor cells is driven by CAFs that are differentially educated by tumor cells in the tumor microenvironment. Our model not only advances our understanding of CAFs' role in tumor progression but also offers an in vitro model for assessing potential therapeutic targets and drug candidates for human gastric cancers. Citation Format: Hyun Ju Hwang, Saet-Byeol Lee, Hyo-Jeong Kuh. Cancer-associated fibroblasts, educated by tumor cells, differentially facilitate cancer cell invasion in cell-line based human gastric tumoroids [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 277.

Abstract 1588: Identification of new signaling pathways between fibroblasts & tumor cells in pancreatic cancer

Abstract We explore the unstudied role of nanoparticles (NPs), recently discovered secreted non-vesicular nanoparticles, in signaling between pre-cancer associated fibroblasts (preCAF), CAFs, and tumor cells. We generated two novel primary lines preCAF PFUM1 from a patient with high-grade intraductal papillary mucinous neoplasms and CAF FUM2 from a patient with pancreatic ductal adenocarcinoma, all tests performed between passages 2 and 3, fibroblast purity validated by RT-PCR and ACTA2 immunofluorescence. Small extracellular vesicles (sEVs), exomeres, and supermeres (the two main forms of NPs) were isolated by the ultracentrifugation method from PF-UM1 and F-UM2 with n=2-3 experiments. To validate the isolation, we used nanoparticle tracking, electron, and fluid-phase atomic force microscopy. Proteins were quantified by BCA assay. Liquid chromatography-mass spectrometry characterized the cargo from each fraction. Human cancer lines MIAPACA2 and SU8686 were treated with 10 μg of each fraction and cell proliferation was tested by phase-contrast images (Cytation 5), analyzed by Ilastik-based 1.4.0 machine-learning. We performed fluorescent Ki-67 and vimentin staining and quantified in QuPath 0.4.3. Non-parametric statistics were performed in Prism GraphPad. NPs constitute the major fraction of small particles released by fibroblasts, containing 80.5% (PFUM1) and 77.5% (FUM2) of the protein load by quantification. Mass spectrometry reveals enrichment of growth factors, epithelial-to-mesenchymal (EMT) transition, and metabolism-associated proteins in the NPs compared to sEVs. At 72 hours, cell proliferation for Su8686 reveals a proliferative effect for all fractions from both PFUM1 and FUM2 (19-63% increase compared to controls, p=0.049 to <0.0001). For MIAPACA2, only supermeres from PFUM1 and FUM2 consistently increased proliferation (9.5-35% increase, p=0.0051 to <0.0001). Ki-67 expression in MIAPACA2, increased by 9.6-23.0% following treatment by NPs (all p<0.0001) and by 2.9-9.2% with sEVs (p=0.011 to <0.0001). EMT was estimated by vimentin in MIAPACA2, as all preCAF fractions from PFUM1 reduced vimentin expression (p<0.001), while CAF FUM2 NPs increased vimentin expression (p<0.0001) while sEVs had no effect (p=0.075). In conclusion, the majority of pancreatic tumor-associated fibroblast small secretome is composed of NPs, exhibiting significant functional pro-tumoral effects. Citation Format: Ayman Al Shoukari, Maelle Batardiere, Nooshin Movahed, Camille beaussier, Jumanah Baig, Melissa gonzalez, Patricia Moraille, Eric Bonneil, Louise Rousseau, Simon turcotte, Kathleen Delgiorno, Marcus Tan, Anna Means, Elham Dianati Ajibisheh, Quoc-huy Trinh. Identification of new signaling pathways between fibroblasts & tumor cells in pancreatic cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1588.

Cancer-oocyte SAS1B protein is expressed at the cell surface of multiple solid tumors and targeted with antibody-drug conjugates

BackgroundSperm acrosomal SLLP1 binding (SAS1B) protein is found in oocytes, which is necessary for sperm-oocyte interaction, and also in uterine and pancreatic cancers. Anti-SAS1B antibody-drug conjugates (ADCs) arrested growth in...

HiHo-AID2: boosting homozygous knock-in efficiency enables robust generation of human auxin-inducible degron cells

Recent developments in auxin-inducible degron (AID) technology have increased its popularity for chemogenetic control of proteolysis. However, generation of human AID cell lines is challenging, especially in human embryonic stem cells (hESCs). Here, we develop HiHo-AID2, a streamlined procedure for rapid, one-step generation of human cancer and hESC lines with high homozygous degron-tagging efficiency based on an optimized AID2 system and homology-directed repair enhancers. We demonstrate its application for rapid and inducible functional inactivation of twelve endogenous target proteins in five cell lines, including targets with diverse expression levels and functions in hESCs and cells differentiated from hESCs.

Diverse temporal and spatial mechanisms work, partially through Stanniocalcin-1, V-ATPase and senescence, to activate the extracellular ATP-mediated drug resistance in human cancer cells.

Resistance to drug therapies is associated with a large majority of cancer-related deaths. ATP-binding cassette (ABC) transporter-mediated drug efflux, epithelial-mesenchymal transition (EMT), cancer stem cells (CSCs), glutathione (GSH), senescence, and vacuole-type ATPase (V-ATPase) all contribute to the resistance. We recently showed that extracellular ATP (eATP) induces and regulates EMT, CSC formation, and ABC transporters in human cancer cells and tumors. eATP also consistently upregulates Stanniocalcin-1 (STC1), a gene that significantly contributes to EMT, CSC formation, and tumor growth. We also found that eATP enhances drug resistance in cancer cells through eATP internalization mediated by macropinocytosis, leading to an elevation of intracellular ATP (iATP) levels, induction of EMT, and CSC formation. However, these factors have never been systematically investigated in the context of eATP-induced drug resistance. In this study, we hypothesized that eATP increases drug resistance via inducing ABC efflux, EMT, CSCs, STC1, and their accompanied processes such as GSH reducing activity, senescence, and V-ATPase. RNA sequencing, metabolomics, gene knockdown and knockout, and functional assays were performed to investigate these pathways and processes. Our study results showed that, in multiple human cancer lines, eATP induced genes involved in drug resistance, elevated ABC transporters' efflux activity of anticancer drugs; generated transcriptomic and metabolic profiles representing a drug resistant state; upregulated activities of GSH, senescence, and V-ATPase to promote drug resistance. Collectively, these newly found players shed light on the mechanisms of eATP-induced as well as STC1- and V-ATPase-mediated drug resistance and offer potential novel targets for combating drug resistance in cancers.

Calendula officinalis - A Crucial and Promising Antifungal Agent with Cytotoxic Biological Attributes

Globally recognized for its antifungal and anti-inflammatory properties, Calendula officinalis (Marigold) comprises various constituents, including carbohydrates, amino acids, terpenoids, glycosides, lipids, fatty acids, and carotenoids. In in-vitro assessments, it demonstrated effectiveness ranging from 54% to 79% against fungal infections, attributed to the significant presence of sesquiterpene, hydrocarbons, sesquiterpenols, δ-cadinene, α-cadinol, and epi-α-muurolol. Conversely, it also showcases robust cytotoxic activity against colon cancer, leukemia, and melanoma cells in human cancer lines, primarily due to the existence of two triterpene glycosides, calenduloside F 6¢-O-n-butyl ester, and calenduloside G 6¢-O-methyl ester.

Near infrared low-level laser effect on electron radiation therapy of non-melanoma human squamous cell cancer: An in-vitro study

PurposeThe aim is to assess the efficacy of combined electron-laser radiation therapy on non-melanoma Squamous cell carcinoma (SCC) cancer cells in an in-vitro model. Material and methodsThe human SCC cancer line (TSCC-1) was purchased from the Iranian Biological Resources Center (IBRC) and cultured according to the standard protocol. The groups of this study were control; only electron radiation; only Low-Level Laser Therapy (LLLT); and combined laser-electron radiation. Irradiation with a 980 nm 50 mW laser was performed in a single session with energy doses of 1 and 1.5 J/cm2. Electron radiation was carried out using a Varian accelerator with a radiation dose of 2 and 4 Gy. 24 h after laser treatment of cancer cells, combined electron irradiation was performed. MTT and flow cytometry bioassays were performed and analyzed to check their effectiveness. ResultsThe cell viability assay for irradiation with 980 nm 50 mW NIR laser in a single session with an irradiation time of 20 and 30 s in combination with 2 and 4 Gy electrons showed a significant difference (P-value <0.01). Flow cytometry test of the electron-radiated samples showed that the sum of early and late apoptosis was the highest. However, for the combination of electron-laser radiation of 30 s with 2 and 4 Gy electrons, the maximum percentage of apoptosis and necrosis was obtained close to the electron alone. Except for the control group, the quantitative interval of induced necrosis of all the groups evaluated by electron alone, laser alone, and combined electron laser was almost equivalent. ConclusionsThis study showed that the physical parameters of a 980 nm NIR LLLT might not have a synergy effect on treatment response in combination with an electron ionizing beam on SCC cancer cells. It is critical to consider the effect of light irradiation parameters for combination therapy in more extensive studies.

NK-92MI Cells Engineered with Anti-claudin-6 Chimeric Antigen Receptors in Immunotherapy for Ovarian Cancer.

Background: The application of chimeric antigen receptor (CAR) NK cells in solid tumors is hindered by lack of tumor-specific targets and inefficient CAR-NK cell efficacy. Claudin-6 (CLDN6) has been reported to be overexpressed in ovarian cancer and may be an attractive target for CAR-NK cells immunotherapy. However, the feasibility of using anti-CLDN6 CAR-NK cells to treat ovarian cancer remains to be explored. Methods: CLDN6 expression in primary human ovarian cancer, normal tissues and cell lines were detected by immunohistochemistry and western blot. Two types of third-generation CAR NK-92MI cells targeting CLDN6, CLDN6-CAR1 NK-92MI cells with domains containing self-activated elements (NKG2D, 2B4) and CLDN6-CAR2 NK-92MI cells with classical domains (CD28, 4-1BB) were constructed by lentivirus transfection, sorted by flow cytometry and verified by western blot and qPCR. OVCAR-3, SK-OV-3, A2780, Hey and PC-3 cells expressing the GFP and luciferase genes were transduced. Subcutaneous and intraperitoneal tumor models were established via NSG mice. The ability of CLDN6-CAR NK cells to kill CLDN6-positive ovarian cancer cells were evaluated in vitro and in vivo by live cell imaging and bioluminescence imaging. Results: Both CLDN6-CAR1 and CLDN6-CAR2 NK-92MI cells could specifically killed CLDN6-positive ovarian cancer cells (OVCAR-3, SK-OV-3, A2780 and Hey), rather than CLDN6 negative cell (PC-3), in vitro. CLDN6-CAR1 NK-92MI cells with domains containing self-activated elements (NKG2D, 2B4) exhibited stronger cytotoxicity than CLDN6-CAR2 NK-92MI cells with classical domains (CD28, 4-1BB). Furthermore, CLDN6-CAR1 NK cells could effectively eliminate ovarian cancer cells in subcutaneous and intraperitoneal tumor models. More importantly, CAR-NK cells combined with immune checkpoint inhibitors, anti-PD-L1, could synergistically enhance the antitumor efficacy of CLDN6-targeted CAR-NK cells. Conclusions: These results indicate that CLDN6-CAR NK cells possess strong antitumor activity and represent a promising immunotherapeutic modality for ovarian cancer.

Combination of shikonin and gefitinib reverses drug resistance in human non-small cell lung cancer and its mechanism

The occurrence and development of tumors are associated with the cell energy metabolism. Inhibiting energy metabolism of lung cancer cells is an important strategy to overcome drug resistance. Based on the cellular energy metabolism pathway, this study observed the effect of combination of shikonin(SKN) and gefitinib(GFB) on the drug resistance in non-small cell lung cancer and explored the underlying mechanism. The human non-small cell lung cancer line HCC827/GR resistant to gefitinib was used as the cell model in vitro. The CCK-8 assay and flow cytometry were employed to investigate the cell viability and apoptosis, respectively. The high performance liquid chromatography was employed to measure the intracellular accumulation of GFB. A Seahorse XFe96 Analyzer was used to detect the changes of cellular energy metabolism. Western blot was employed to determine the expression of the proteins involved in the drug resistance. The tumor-bearing nude mouse model was used to verify the efficacy of SKN+GFB in overcoming drug resistance in vivo. The results showed that SKN+GFB significantly reduced the IC_(50) of GFB on HCC827/GR cells, with the combination index of 0.628, indicating that the combination of the two drugs had a synergistic effect and promoted cell apoptosis. SKN increased the intracellular accumulation of GFB. SKN+GFB lowered the oxygen consumption rate(OCR) and glycolytic proton efflux rate(GlycoPER) in cell energy metabolism, and down-regulated the overexpression of PKM2, p-EGFR, P-gp, and HIF-1α in drug resistance. The results of reversing drug resistance test in vivo showed that GFB or SKN alone had no significant antitumor effect, while the combination at different doses induced the apoptosis of the tumor tissue and inhibited the expression of PKM2 and P-gp, demonstrating a significant antitumor effect. Moreover, the tumor inhibition rate in the high-dose combination group reached 64.01%. In summary, SKN+GFB may interfere with the energy metabolism to limit the function of HCC827/GR cells, thus reversing the GFB resistance in non-small cell lung cancer.

EFFECT OF SPERMININE AND ITS COMBINATION WITH SPERMINOXIDASE INHIBITORS ON THE PROFILE OF POLYAMINE AND SURVIVAL OF HUMAN PROSTATE CANCER CELLS

Summary. Aim: to investigate the effect of spermine (Spn) and spermine oxidase inhibitors (SMO) and their effect in combination on the polyamine profile in human PCa cell lines and to compare the nature of polyamine changes with tumor cell viability. Object and methods: the study was performed on cell cultures of hormone-sensitive (LNCaP) and hormone-resistant (DU-145) cell lines. The profile of polyamines in tumor cells and the activities of spermine oxidase and polyamine oxidase (PAO) were determined by high-performance liquid chromatography. Results: the addition of Spn to the culture medium causes a significant accumulation of this polyamine in cells, a decrease in putrescine (Put) and spermidine (Spd). The use of SMO inhibitors (chlorhexidine and MDL-72527) is accompanied by a decrease in Spd levels and accumulation of acetylated spermidine (AcSpd). The use of Spn leads to a decrease in the proliferation rate — the molar ratio of spermidine/spermine (Spd/Spn). Particularly low values of the Spd/Spn ratio were observed when spermine was used in combination with SMO inhibitors. When Spn was used in combination with SMO inhibitors, the lowest cell viability rates were also observed, indicating a more effective inhibition of cell growth with the combined use of these factors compared to their use separately, even in significantly higher concentrations. The use of chlorhexidine and chlorhexidine together with spermine led to a decrease in the activity of SMO and PAO. Conclusions: the use of spermine and spermine in combination with SMO inhibitors leads to changes in the polyamine profile in tumor cells characteristic of growth inhibition (decrease in Spd, Put and increase in Spn) and a decrease in the molar ratio Spd/Spn. These changes are accompanied by a decrease in the survival of cells of hormone-sensitive (LNCaP) and hormone-resistant (DU-145) human prostate cancer lines. The obtained results show the prospects for further study of spermine and its combination with SMO inhibitors as potential agents for the treatment of prostate cancer.

Strong in vitro anticancer activity of copper(ii) and zinc(ii) complexes containing naturally occurring lapachol: cellular effects in ovarian A2780 cells.

Copper(ii) and zinc(ii) complexes with lapachol (HLap) of the composition [M(Lap)2(N-N)] and [Cu(Lap)(H2O)(terpy)]NO3 (4), where M = Cu (1-3) or Zn (for 5-7), and N-N stands for bathophenanthroline (1 and 5), 5-methyl-1,10-phenanthroline (2 and 6), 2,2'-bipyridine (3), 2,2';6',2''-terpyridine (terpy, 4) and 1,10-phenanthroline (7), were synthesised and characterised. Complexes 1-5 revealed strong in vitro antiproliferative effects against A2780, A2780R, MCF-7, PC-3, A549 and HOS human cancer lines and MRC-5 normal cells, with IC50 values above 0.5 μM, and reasonable selectivity index (SI), with SI > 3.8 for IC50(MRC-5)/IC50(A2780). Considerable time-dependent cytotoxicity in A2780 cells was observed for complexes 6 and 7, with IC50 > 50 μM (24 h) to ca. 4 μM (48 h). Cellular effects of complexes 1, 5 and 7 in A2780 cells were investigated by flow cytometry revealing that the most cytotoxic complexes (1 and 5) significantly perturbed the mitochondrial membrane potential and the interaction with mitochondrial metabolism followed by the triggering of the intracellular pathway of apoptosis.