HeLa Research Articles

Promyelocytic leukemia protein (PML) knockout increases mitochondrial Ca2+ uptake in HeLa cells

The multifunctional promyelocytic leukemia protein (PML) is involved in the regulation of various cellular processes in both physiological and pathological conditions. Specifically, PML is one of the inositol-1,4,5-trisphosphate receptors (IP3Rs) activity regulators and can influence Ca2+ transport from the endoplasmic reticulum (ER) to mitochondria. In this work, the effects of PML knockout on calcium homeostasis in the cytosol, ER, and mitochondria of HeLa cells were studied upon stimulation with histamine, which induces Ca2+ mobilization from the ER via IP3Rs. We utilized calcium indicators with different subcellular localizations, including synthetic dyes Fura-2 (cytosolic), Xrhod-5F (mitochondrial), and protein sensor R-CEPIAer (ER), as well as mitochondrial potential-sensitive probes Rh123 and TMRM. Our results show that PML knockout induced changes in HeLa cell and mitochondrial morphology, slightly decreased basal and integral Ca2+ levels, enhanced mitochondrial Ca2+ uptake from the cytoplasm, and maintained residual mitochondrial potential after depolarization. Additionally, it reduced the Ca2+ pool in ER membranes not associated with histamine receptor activation and, consequently, IP3Rs. These findings suggest that changes in calcium ion transport due to PML knockout in HeLa cells affect mitochondrial activity.

RETRACTION: Targeted Delivery of Doxorubicin in HeLa Cells Using Self‐Assembled Polymeric Nanocarriers Guided by Deep Eutectic Solvents

AbstractRETRACTION: P. Pradeepkumar, M. Rajan, H. S. Almoallim, and S. A. Alharbi, “Targeted Delivery of Doxorubicin in HeLa Cells Using Self‐Assembled Polymeric Nanocarriers Guided by Deep Eutectic Solvents,” ChemistrySelect 6, No. 28 (2021): 7232–7241, https://doi.org/10.1002/slct.202000036.The above article, published online on July 26, 2021, in Wiley Online Library (wileyonlinelibrary.com), has been retracted by agreement between the journal editor‐in‐chief, Preeti Vashi, and Wiley‐VCH GmbH.The retraction has been agreed upon following an investigation into concerns raised by a third party, which identified duplicated images in Figures 7A, S3C, and S5A–D. The investigation also identified image duplication between Figure 6A and a figure from a previously published article by a similar group of authors. The authors were unable to provide a satisfactory explanation and did not provide all the original data. As a result, the data and the conclusions are considered unreliable.

β-amyrin heptadecanoate, a new oleanane triterpenoid with α-glucosidase inhibitory and cytotoxic activities from the leaves of Averrhoa bilimbi L.

A previously unreported oleanane triterpenoid, β-amyrin heptadecanoate (1), was isolated from the leaves of Thai Averrhoa bilimbi (Oxalidaceae), along with five known compounds, β-amyrin (2), β-sitosterol (3), β-sitosterol-D-glucoside (4), β-sitosteryl oleate (5), and α-tocopherol (6). Their structures were elucidated through spectroscopic analysis, including extensive NMR and HRESIMS, and by comparison with the previous literature. All isolated compounds were evaluated for their α-glucosidase inhibitory and cytotoxic activities. Compound 6 exhibited the highest α-glucosidase inhibition (IC50 of 0.72 ± 1.02 µM), significantly outperforming the standard acarbose (IC50 of 82.00 ± 0.24 μM). Furthermore, compound 4 demonstrated the most potent cytotoxicity against HeLa cells (IC50 of 34.92 ± 0.45 μM), while compound 3 exhibited the strongest cytotoxicity towards A549 cells (IC50 of 9.17 ± 0.30 μM). Additionally, a molecular docking study was conducted on the active α-glucosidase inhibitors to estimate their binding affinities and to identify the ligand-binding sites within the enzyme.

Chitosan-naphthalimide probes for dual channel recognition of HClO and H2S in cells and their application in photodynamic therapy

The combination of bio-imaging with photodynamic therapy (PDT) to accomplish theranostics is promising in cancer treatment. Three chitosan-naphthalimide probes were studied in this work. 4-(5-Bromothiophen-2-yl)-1,8-naphthalic anhydride was first synthesized, and then reacted with chitosan to obtain the macromolecules (CS-N-Br). The recognition group thiomorpholine or its derivatives were introduced into CS-N-Br to obtain nano-probes (CS-N-ML, CS-N-BSZ, CS-N-FSQ) eventually. The studies revealed that CS-N-ML and CS-N-FSQ exhibit high selectivity and can specifically recognize HClO and H2S. CS-N-ML and CS-N-FSQ can perform exogenous and endogenous confocal imaging of HClO and H2S in cells also. CS-N-ML's ability to target lysosomes positions indicated it could act as a lysosome-specific probe. It was discovered that the probes generate superoxide anions (O2•−) via a Type I mechanism. This discovery endows the probes with high photosensitizing activity even under hypoxic conditions. There is a positive correlation between the extent of the conjugated system and the photosensitivity of the probes, indicating that an enhanced conjugation leads to increased photosensitivity. Upon light irradiation, the probes generate ROS within HeLa cells. These results suggested that these probes can achieve theranostics for diseases associated with abnormal levels of HClO and H2S.

Biosynthesis of Chitosan encapsulated Silver- Nanoparticles using Probiotic-Lactobacillus plantarum strain and It's In vitro Anticancer Assessment on HeLa cells

Biosynthesis of Chitosan encapsulated Silver- Nanoparticles using Probiotic-Lactobacillus plantarum strain and It's In vitro Anticancer Assessment on HeLa cells

Engineering a fluorescent probe for the visual and wearable detection of N2H4 in foods, environment samples and biological imaging

Hydrazine (N2H4), as an important chemical raw material widely used in many fields, caused serious harm to human, food and environment. Therefore, a smart fluorescent probe TPC-N2H4 with intramolecular charge transfer (ICT) process was fabricated for colorimetric and fluorescent differentiating of N2H4 in many samples. When TPC-N2H4 met N2H4, blue emission at 451 nm was obviously observed attributed to the interruption of ICT process, resulting in the formation of the hydrazone structure. Additionally, smartphone-assisted colorimetric and fluorescent sensing platforms had been developed for real-time monitoring of N2H4. Importantly, TPC-N2H4 with low toxicity revealed its ability to image N2H4 in HeLa cells, onion and Arabidopsis thaliana. Interestingly, a hydrogel-coated cotton swab sensor was developed with TPC-N2H4 as recognition unit, which showed obvious color and emission changes after N2H4 fumigation with fast response speed. Flexible fluorescent TPC-N2H4-glove was constructed for wearable N2H4 detection and used to monitor N2H4 in food and environment samples. In brief, the TPC-N2H4 probe not only showed great potential in N2H4 detection for environmental and food safety, but also provided a new method for wearable tracking platforms in toxic substance detection.

Quercetin exhibits cytotoxicity in cancer cells by inducing two-ended DNA double-strand breaks.

Quercetin, a flavonoid, is involved in the induction of DNA double-strand breaks (DSBs), in addition to its antioxidant properties. Although DNA topoisomerase II (Top2) and reactive oxygen species (ROS) have been suggested as possible mechanisms through which quercetin induces DSBs, the exact mechanism remains unclear. In this study, we examined the mechanism of DSB induction by quercetin and its repair using HeLa cells and gene-knockout cell lines generated from human Nalm-6cells. Immunofluorescence staining for γH2AX, a DSB marker, and analysis of the frequency of random integration of foreign DNA, which correlates with the number of DSBs and DSB repair pathways, indicated that quercetin induces DSBs in a concentration-dependent manner. The sensitivity assay suggested that the factor involved in quercetin-induced DSBs was not Top2. However, ROS was found to accumulate transiently in quercetin-treated HeLa cells. Furthermore, the addition of ascorbic acid increased the survival of quercetin-treated HeLa cells, suggesting that quercetin induces a transient accumulation of ROS, which in turn induces DSBs. The resulting DSBs were repaired primarily by non-homologous end-joining and homologous recombination, similar to X-ray-induced DSBs. Taken together, quercetin, used as a radiomimetic agent, has the potential to produce effects equivalent to those of an X ray-dose at a relatively low risk.

Innovative anticancer nanocomposites from Corchorus olitorius mucilage/chitosan/selenium nanoparticles

Cancers are continuing to threaten human health globally; the achievement of effectual and biosafe anticancerous compounds is a precious goal. The extraction of Corchorus olitorius mucilage (Jm) and its usage for selenium nanoparticles (SeNPs) biosynthesis was projected. The innovative formulation of bioactive nanocomposites (NCs) from Jm/SeNPs and chitosan nanoparticles (Cht) was also proposed to apply these NCs as effectual anticancers against CaCo-2 and HeLa cancerous cells. The Jm/SeNPs biosynthesis (mean diameter = 6.45 nm) was innovatively achieved and confirmed using infrared and ultraviolet-visible analysis. The constructions of different NCs were done (N1: 2Jm/SeNPs:1Cht; N2: 1Jm/SeNPs:1Cht; and N3: 1Jm/SeNPs:2Cht) with mean particles' diameter of 88.41, 46.86 and 69.35 nm, respectively. The cytotoxicity assay of constructed NCs indicated their potentialities to suppress examined cells; N1 (negatively charged; −16.2 mV) was the most forceful with IC50 of 12.36 and 73.15 mg/L against CaCo-2 and HeLa cells, respectively. The scanning microscopy imaging of treated CaCo-2 cells with N1 of Cht/Jm/SeNPs indicated that the NCs led to remarkable apoptotic destructions of treated cells, including cell shrinkage, membrane blebbing, cytoplasmic vacuolization, cell debris and apoptotic indices. The innovative NCs from Cht/Jm/SeNPs are promisingly recommended as effectual, natural and bioactive anticancer formulations against human cancers.

Novel quinoline-based chemosensor as specific fluorescence sensing of copper ions in cancer cells and for organelle-specific imaging application

This work reports a simple synthesis of a quinoline-based organic molecule, (E)-N′-((6-methoxy-2-oxo-1,2-dihydroquinolin-3-yl)methylene) picolinohydrazide (PHMQ), and its ability to distinguish Cu2+ ion. Through the use of spectroscopic and photometric titration techniques, PHMQ’s selectivity toward various cations and anions was examined. In just two minutes, PHMQ showed fast detection of Cu2+ ions, and clear spectral fluctuations proposed persistent PHMQ-Cu2+ complex formation. The calculated emission quenching constant (1.46 × 105 L/mol) and binding constant (1.25 × 104 L/mol) values suggested that strong emission quenching occurs between the PHMQ and Cu2+ ions. A linear reduction in fluorescence response was noted for Cu2+ ions concentrations between 2–26 μM, with a detection limit of 43.4 nM. Job’s plot, ESI-mass, NMR titration experiments, and density functional theory (DFT) simulations established a 1:1 stoichiometry for the PHMQ-Cu2+ complex. Additionally, PHMQ was efficaciously utilized to quantify Cu2+ ions in drinking and tap water samples and HeLa cells, illustrating durable cellular penetrability and a fluorescence ’Turn-Off’ response to Cu2+ ions. This sensor also established usefulness in mitochondrial-targeted imaging. For the detection of Cu2+ ions in biological and environmental environments, PHMQ is a promising detector due to its high sensitivity, selectivity, and quick response.

Investigation of the Efficacy of Nowarta110 in the Treatment of HPV-16 and HPV-18 Oncogenically-transformed Human Cells and Cancer-implanted Animal Models.

Cervical cancer is the third leading cause of cancer-related death in women worldwide. Nearly all cases of cervical cancer are due to infection with human papillomavirus (HPV). Nowata110 has shown breakthrough therapeutic efficacy in phase II clinical study against plantar warts, with no reported side effects. This study evaluated the effectiveness of Nowarta110 in killing cultured HPV-transformed cancer cells and its anti-cancer effects in mice, using both vaginally engrafted and subcutaneously implanted animal cancer models. Nowata110 is a novel compound composed of colloidal silver and fig extract. The cytotoxic properties of Nowarta110 were evaluated on HPV-16 and HPV-18-transformed human cancer cells (ARPE-19/HPV-16 and HeLa cells, respectively). The therapeutic potential of Nowarta110 in vivo was evaluated following the engraftment of ME-180 epidermoid carcinoma cells in the mouse vagina or their subcutaneous implantation. Nowarta110 treatment was initiated when the tumor reached an approximate volume of 65 mm3 Results: Our data showed that HPV16-expressing human retinal pigmented epithelial cells are susceptible to Nowarta110-induced cell death, with a reduction in cell viability observed at 1% Nowarta110 and a complete absence of viable cells at 5% Nowarta110. In HPV18-expressing HeLa cells, Nowarta110 caused significant and rapid cell death at a dose of 5%, whereas lower doses of 1% did not produce the same effects. The results from animal studies indicated that Nowarta110 effectively induced tumor regression in both the intravaginal and subcutaneous tumor models. Nowarta110 effectively induced cell death in HPV-16 and HPV-18-transformed human cancer cells. It also effectively induced tumor regression in mouse models with intravaginally engrafted or subcutaneously implanted cancer cells. Considering the high prevalence of HPV-induced cervical dysplasia and cancer and the lack of treatment, clinical studies to promote the implementation of Nowarta110 are warranted.

Synthesis, Docking, and Dynamic Studies of New Pyrano‐[3,2‐e]‐Pyrazole Fused 1,2,4‐Triazolo‐[4,3‐c]‐pyrimidine Analogues as Anticancer Agents

AbstractA novel series of dihydropyrazolo‐[4′,3′:5,6]‐pyrano[3,2‐e][1,2,4]triazolo[4,3‐c]‐pyrimidine analogues were designed, synthesized, and their cytotoxic potential against four tumor cell lines assessed. HRMS, 1H NMR, and 13C NMR spectra analysis were used to describe the novel compounds. Compound 9c had noteworthy anticancer activity in vitro against the Hela, MCF‐7, A549, and A2780 cell lines, with IC50 values of 0.98 ± 0.26, 3.11 ± 0.18, 1.24 ± 0.88, and 2.62 ± 0.56 µM, in that order. The molecular docking and dynamics simulation studies revealed the lead molecule's action mechanism with EGFR target protein.

Evaluation of safety and biomedical potential of water-soluble oat lignin Avena sativa L.

The study of the value of lignin for biomedical use is generating growing interest. For the first time, the safety and biological efficacy of lignin from the stems of the oat Avena sativa L. were studied, necessary for a preliminary assessment of its biomedical potential, have been studied. In vitro experiments, a sample of oat lignin exhibited cytotoxicity to the HeLa, A549, and HT-29 cancer cell lines, depending on the concentration. At maximum concentrations 125 and 150 μg/ml, it reduced their survival and increased the level of reactive oxygen species. In vivo experiments, a sample of oat lignin, with acute (from 5 to 250 mg/kg body weight) and chronic (300, 1200 and 2000 mg/kg body weight) administration, did not have a toxic or genotoxic effect on the organs of mice. The biological efficacy of the oat lignin was manifested in activation of repair processes in bone marrow and thyroid gland, a decrease in the level of abnormal spermatozoa in males, stimulation of reproductive performance of females and in increase in research activity and a decrease in the level of anxiety in animals. The results indicate the prospects for further study of the medical and biological potential lignin of the oat.

Novel benzenesulfonamide-aroylhydrazone conjugates as carbonic anhydrase inhibitors that induce MAPK/ERK-mediated cell cycle arrest and mitochondrial-associated apoptosis in MCF-7 breast cancer cells

A series of novel 4-alkylthio-2-chloro-5-[(2-arylmethylidene)hydrazinecarbonyl]benzenesulfonamide derivatives 3–22 were synthesized and evaluated for their inhibitory activity against human carbonic anhydrase isozymes hCA I, hCA II, hCA IX, and hCA XII. These compounds showed varying degrees of activity against the studied isoenzymes. However, the importance of substituent choice in designing potent carbonic anhydrase inhibitors is highlighted by the strong inhibition profiles of compounds 3 and 10 against hCA IX and the low average KI values for compounds 9 and 10 (134 nM and 77 nM, respectively). All the synthesized compounds were evaluated for their antiproliferative activity toward HeLa, HCT-116, and MCF-7 cell lines. Compounds 9 and 19 exhibited significant activity, particularly against the MCF-7 cell line (IC50 values of 4 μM and 6 μM, respectively). Notably, compound 9 demonstrated a high selectivity index (SI = 8.2) for MCF-7 cells. The antiproliferative effects of compounds 9 and 19 were linked to the induction of cell cycle arrest and apoptosis via the mitochondrial pathway and involved the activation of the MAPK/ERK signaling pathway. Inhibition of MAPK/ERK activity reduced the compounds’ ability to induce cell cycle arrest and apoptosis, indicating the critical role of this pathway. These findings suggest that compounds 9 and 19 are promising candidates for further development as specific and potent anticancer agents targeting the MAPK/ERK pathway.

Discovery and Development of HDAC Inhibitors: Approaches for the Treatment of Cancer a Mini-review.

Histone deacetylase (HDAC) inhibitors have emerged as promising cancer therapeutics due to their ability to induce differentiation, cell cycle arrest, and apoptosis in cancer cells. In the present review, we have described the systemic discovery and development of HDAC inhibitors. Researchers across the globe have identified various small molecules like benzo[d][1,3]dioxol derivatives, belinostat analogs, pyrazine derivatives, quinazolin-4-one-based derivatives, 2,4-imidazolinedione derivatives, acridine hydroxamic acid derivatives, coumarin derivatives, tetrahydroisoquinoline derivatives, thiazole-5-carboxamide, salicylamide derivatives, β-peptoid-capped HDAC inhibitors, quinazoline derivatives, benzimidazole and benzothiazole derivatives, and β- elemene scaffold containing HDAC inhibitors. Most of the scaffolds have shown attractive IC50 (μM) in various cell lines like HDAC1, HDAC2, HDAC6, PI3K, HeLa, MDA-MB-231, MCF-10A, MCF-7, U937, K562 and Bcr-Abl cell lines.

RecQ protein-like 4 drives cisplatin chemosensitivity of cervical cancer cells by modulating annexin A2.

Cervical cancer is a common malignant tumor in women with high morbidity and mortality. Chemotherapy drugs such as cisplatin (DDP) are easy to cause chemotherapy resistance and affect the therapeutic effect. Hence, it is critical to design new therapies that can reverse chemotherapy resistance and increase sensitivity to chemotherapy drugs. This study investigated the function of RecQ protein-like 4 (RECQL4) in DDP-resistant cervical cancer cells and its regulatory mechanism. By constructing DDP-resistant Hela and CaSki cell lines, it was found that RECQL4 expression was elevated. RECQL4 knockdown is able to promote apoptosis, DNA damage, and increase the DDP sensitivity in cervical cancer cells. In vivo experiments have demonstrated that knockdown of RECQL4 suppresses tumor growth and promotes tumor apoptosis. Next, we investigated the potential regulatory relationship of RECQL4 to Annexin A2 (ANXA2). The results demonstrated that RECQL4 binds to ANXA2. Knockdown of RECQL4 downregulates the ANXA2 expression via promoting ubiquitination. Furthermore, we also found that ANXA2 overexpression partially abolished the role of RECQL4 knockdown in promoting apoptosis and DNA damage of cervical cancer cells, suggesting that RECQL4 plays a role in DDP sensitivity of cervical cancer cells by mediating ANXA2. In conclusion, the present study suggests that knocking down RECQL4 reduces DDP sensitivity in cervical cancer cells by modulating ANXA2. Targeting RECQL4 therapy may be a new strategy to improve chemosensitivity of cervical cancer cells.

Facile synthesis of Fe3O4 photothermal nanoparticles coated with poly-3,4-dihydroxybenzaldehyde for improved photothermal therapy of cancer cells

Given the significant rise in the number of individuals diagnosed with cancer, the management of cancer therapy has become a prominent concern in society and a formidable task in human health care. Photothermal therapy has garnered significant interest in cancer treatment because of its exceptional efficacy and minimal invasiveness. Thus, current research mainly focuses on developing biocompatible materials with excellent photothermal properties. Therefore, we synthesized Fe3O4 nanoparticles coated with poly-3,4-dihydroxybenzaldehyde (Fe3O4@PDBA) where 3,4-dihydroxybenzaldehyde was covalently polymerized onto the surface of Fe3O4. The induction of PDBA leads to a dramatical improvement in the photothermal properties of Fe3O4. Besides, the thickness of the PDBA shell layer is also a key factor in the photothermal performance and various physicochemical properties of Fe3O4@PDBA and the photothermal performance of Fe3O4@PDBA increases with increasing thickness of the PDBA shell. Additional structural characterization of the products, DFT calculations of the UV-Vis absorption spectra and the HOMO-LUMO gap under the simple model of the polymer, and product identification by the use of H1NMR, MS, and other techniques were performed. Through multiple characterizations, the reaction solution color and UV-Vis absorption spectra were essentially consistent with the computed findings. The calculations confirm that the polymerization process of PDBA is progressively red-shifted and the absorption is gradually enhanced in the NIR region. Fe3O4@PDBA exhibited strong cytotoxicity against cancer cells (Hela, 98.0%) in vitro photothermal treatment. Hence, Fe3O4@PDBA exhibits superior photothermal characteristics, targeting, biocompatibility (99.0%), and degradability, which brings about significant potential for photothermal cancer therapy.

Icariin ameliorates Coxsackievirus B3-induced viral myocarditis by modulating the S100 calcium binding protein A6/β-catenin/c-Myc signaling pathway

BackgroundCoxsackievirus B3 (CVB3) is a leading cause of viral myocarditis and is currently lacking specific pharmacological treatments, highlighting the critical need for therapeutic development. Icariin (ICA), a prenylated flavonol glycoside, was previously found to exhibit several pharmacological effects, but its potential to combat CVB3 remains uninvestigated. PurposeThis study aimed to elucidate the anti-CVB3 efficacy of ICA and elucidate its molecular mechanisms. MethodsCVB3-infected HeLa cells, H9C2 cells and neonate rat ventricular cardiomyocytes (NRVCs) were selected as in vitro models, and were treated with ICA at 1 and 10 μM. Additionally, BALB/c mice that were infected with CVB3 via intraperitoneal injection were chosen as in vivo model and were treated with ICA or ribavirin over 3 days. The effect of ICA against CVB3 was determined by Cell Counting Kit-8 (CCK-8) assay, western blot, real-time fluorescence quantitative PCR (RT–qPCR), terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay, hematoxylin and eosin (H&E) staining, immunohistochemistry (IHC) and flow cytometry. ResultsIn this study, it was found that ICA is capable of reducing CVB3 viral load both in vitro and in vivo. Mechanistic studies suggested that ICA prevents cardiomyocyte apoptosis by attenuating the S100 calcium binding protein A6 (S100A6)/β-catenin/c-Myc signaling pathway. Additionally, ICA inhibits the secretion of proinflammatory cytokines tumor necrosis factor alpha (TNF-α), interleukin-1beta (IL-1β) and CXC motif chemokine ligand 2 (CXCL2) in heart tissue, thereby mitigating CVB3-induced myocarditis. Moreover, ICA also regulates the immune response of CD4+ T, CD8+ T and Treg cells by changing the cells numbers in spleen tissue. Lastly, ICA can reduce the load of other enteroviruses (such as CVA6, CVA16 and EV71) in rhabdomyosarcoma (RD) cells as well. ConclusionOur findings indicate that ICA provides significant protection against CVB3 infection by modulating the S100A6/β-catenin/c-Myc signaling pathway, suggesting its potential use as a novel drug against CVB3 infection in clinical application.

Green approach of cobalt sulfide nanoparticles from novel red stigma of Crocus sativus and multifaceted biomedical advancement

The present study was based on green protocols for synthesizing cobalt sulfide nanoparticles using saffron stigma flower extract (Crocus sativus), as saffron is considered a potent traditional medicine. Furthermore, the novel cobalt sulfide (CoS) nanoparticles prepared by the green approach for the first time with saffron improved showed cytotoxicity and antibacterial activity, as well as significant antitumor activity of Hela, A549, and MCF 7 cells viz in vitro studies. The saffron-cobalt sulfide nanoparticles underwent characterization using various techniques which include the X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), Advances in scanning electron microscopy (SEM), and high-resolution transmission electron microscopy (HR-TEM), and the energy dispersive X-ray analysis spectroscopy. Both XRD, and FTIR confirmed the synthesis of pure cobalt sulfide nanoparticles. The resulting saffron-based cobalt sulfide nanoparticles exhibited a nanosphere structure, as confirmed by HRTEM, indicating the formation of nanosphere and microstructures. In vitro studies were conducted to assess the cytotoxicity and cell viability of Hela cells, A549, and MCF-7 cells. And, antibacterial studies were performed against various bacterial strains using different concentrations of cobalt sulfide nanoparticles derived from the red stigma of saffron (Crocus sativus) only the red stigma extract. The MTT assay analysis, cell viability, and morphological studies demonstrated the cytotoxic studies of the synthesized cobalt sulfide nanoparticles on cancer cells. The antibacterial test of the nanoparticles was also evaluated, revealing significant zones of inhibition (mm) against pathogens and cobalt sulfide nanoparticles using saffron extract, showcasing their potent cytotoxic, antibacterial, and antitumor activities with promising biomedical applications.

Identification and validation of miR-21 key genes in cervical cancer through an integrated bioinformatics approach

Cervical cancer is one of the most prevalent female reproductive cancers. miR-21 is a multi-target oncomiR that has shown its potential in regulating several cancers including colon, pancreatic, breast, prostate, ovarian, and cervical cancer. However, the signaling network of miR-21 remains underexplored, and only a limited number of miR-21 gene targets in cervical cancer have been reported. In this context, the present study was undertaken to evaluate the role of miR-21 in cervical cancer by combining in silico analysis with in vitro validation in cervical cancer cells. The miR-21 target genes were predicted using four different prediction tools: miRwalk, DIANA, miRDBs, and TargetScan. A total of 113 overlapping target genes, common in at least three of the prediction tools, were shortlisted and subjected to functional enrichment analysis. The analysis predicted that JAK-STAT, MAPK, neurotrophin, and Ras signaling pathways are significantly (p≤0.05) targeted by miR-21. The MCODE plugin identified the potential cluster in the protein-protein interaction network based on the highest degree of connectivity. After GEPIA2 validation of all 20 hub genes, NTF3, LIFR, and IL-6R were shortlisted for validation in cervical cancer cell lines. The results showed that NTF3, LIFR, and IL-6R were significantly upregulated in the miR-21 knockdown CaSki cell lines in 6.27, 1.92 and 1.71 folds (p≤0.01), respectively. Similarly, in HeLa cell lines expression of NTF3, LIFR, and IL-6R were overexpressed in 4,06, 5.65, 2.42 folds (p≤0.001), respectively. confirming the role of miR-21 in regulating the expression of these genes. Additionally, the knockdown of miR-21 significantly inhibited the secretion of matrix metalloproteinases by CaSki cells. These results highlight that miR-21 could be a potential therapeutic target for cervical cancer, although further preclinical and clinical studies are required to validate its role and efficacy.

Gradient-Elution Nanoflow Liquid Chromatography without a Binary Pump: Smoothed Step Gradients Enable Reproducible, Sensitive, and Low-Cost Separations for Single-Cell Proteomics

Mass spectrometry-based proteome profiling of trace analytes including single cells benefits from liquid chromatography separations operated at low flow rates (e.g., <50 nL/min). However, high-pressure binary pumps needed to achieve such flow rates are not commercially available, and instead require splitting of the gradient flow to achieve low-nanoliter-per-minute flow rates. Gradient flow splitting can waste solvent and lead to flow inconsistencies. To address this, we have developed a method for creating gradients by combining plugs of mobile phase of increasing solvent strength together in a column, and then relying on Taylor dispersion to form the desired smooth gradient profile. Additionally, our method dramatically reduces costs, as only a single isocratic high-pressure pump is required. Following development of gradient profiles for both 10- and 20-min active gradients, we measured 200 pg injections of HeLa digest using a timsTOF mass spectrometer. Finally, we investigated differences in protein expression between single cells originating from two different colonies of ATG-knockout HeLa cells. Thousands of proteins were quantified, and a potential mechanism explaining differential immune responses of these two colonies upon exposure to viral DNA treatment was determined.