SKOV3 Cell Lines Research Articles

Suitable reference gene for silencing methods using microRNA encapsulated nanoparticles chitosan for the ovarian cancer cell line

MicroRNAs (miRNAs) are key regulators of gene expression, and their accurate quantification is essential for elucidating miRNA functional roles. Quantitative reverse transcription PCR (qRT-PCR) is widely utilized for miRNA expression analyses, although appropriate data normalization is critical for generating reliable results. The present study aimed to identify a suitable reference gene for normalizing miRNA qRT-PCR data in the ovarian cancer cell line SKOV-3. SKOV-3 cells were treated with miRNA-loaded chitosan nanoparticles or left untreated as a control. Total RNA, including small RNAs, was extracted from SKOV-3 cells and assessed for quality. The expression stability of 15 candidate reference genes was evaluated by GeNorm analysis. miR-584-5p was identified as the most stably expressed reference gene. To validate miRNA profiling, miR-584-5p was used to normalize expression levels, revealing distinct patterns of differential miRNA expression. Our findings provide critical insights into aberrant miRNA expression in ovarian cancer cells and underscore proper normalization as essential for accurate quantification of miRNAs by qRT-PCR.

MUM1L1 as a Tumor Suppressor and Potential Biomarker in Ovarian Cancer: Evidence from Bioinformatics Analysis and Basic Experiments.

Ovarian cancer (OC) is the most prevalent gynecologic malignancy, with high mortality rates. However, its pathogenesis remains unclear. The current study aimed to explore potential biomarkers and suppressor genes for diagnosing and treating OC. Biochemical and bioinformatics approaches were used to detect differentially expressed genes (DEGs) in ovarian tissues via integration analysis. Kaplan-Meier plot analysis was performed to assess progression-free survival and overall survival according to DEGs. Then, we constructed a protein-protein interaction (PPI) network based on data from the STRING database to identify the related target genes of DEGs. Finally, DEGs regulating the proliferation, migration, and invasion of SKOV3 cell lines were validated via in vitro experiments. Four DEGs (MUM1L1, KLHDC8A, CRYGD, and GREB1) with enriched expression in ovarian tissues were explicitly expressed in the ovary based on an analysis of all human proteins. MUM1L1 had high specificity, and its expression was higher in normal ovarian tissues than in OC tissues. Kaplan-Meier plot analysis showed that a high MUM1L1 expression was associated with longer progression-free survival and overall survival in OC. Based on the PPI analysis results, CBLN4, CBLN1, PTH2R, TMEM255B, and COL23A1 were associated with MUM1L1. In vitro studies revealed that MUM1L1 overexpression decreased the proliferation, migration, and invasion ability of SKOV3 cell lines. Meanwhile, MUM1L1 knockdown had contrasting results. MUM1L1 is a tumor suppressor gene and is a potential biomarker for diagnosing and treating OC.

Mice ovarian microbiome investigation divulges prospective fount of anticancer and antimicrobial metabolites

The impact of bacterial microbiome metabolites on human health, most particularly on the prevention and treatment of cancer, has gained significant interest in recent times. The study focused on the isolation of bacterial flora associated with the ovaries of 8-week-old mice and examined its ability to produce metabolites with potential antibacterial, antioxidant, and anticancer properties. The isolates were identified based on biochemical and 16S rRNA sequencing. The ethyl acetate extract from isolates was screened for antimicrobial and antioxidant capacity. The 3-[4,5-dimethylthiazole-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay was used to evaluate the cytotoxicity of the selected isolate against the ovarian carcinoma cell lines SKOV-3 and PA-1. The ethyl acetate extract from Bacillus velezensis OM03 exhibited significant antibacterial activity against all the tested bacteria, with a MIC value ranging from 50 to 100 µg/mL. Furthermore, the extracts demonstrated hydrogen peroxide and 2, 2-diphenyl-1-picrylhydrazyl (DPPH) scavenging activities at rates of 88.50% and 87.78%, respectively. The extracts displayed substantial concentration-dependent antiproliferative/cytotoxic activities against SKOV-3 and PA-1 cell lines after 24 and 48 h of treatments, respectively. Further chemical analysis of the extract using HR-LCMS revealed the presence of bioactive compounds such as myriocin, 2,3-diethylpyrazine, dihydrodeoxystreptomycin, cyclo (L-Phe-L-Pro), C16 sphinganine, and other twenty-nine compounds that have been formerly reported and are accountable for the targeted activities. Bacillus velezensis OM03 may be further investigated for the creation of novel therapeutics, particularly for the treatment of ovarian cancer.

Analytical Validation of a Spiral Microfluidic Chip with Hydrofoil-Shaped Pillars for the Enrichment of Circulating Tumor Cells.

The isolation of circulating tumor cells (CTCs) from peripheral blood with high efficiency remains a challenge hindering the utilization of CTC enrichment methods in clinical practice. Here, we propose a microfluidic channel design for the size-based hydrodynamic enrichment of CTCs from blood in an epitope-independent and high-throughput manner. The microfluidic channel comprises a spiral-shaped part followed by a widening part, incorporating successive streamlined pillars, that improves the enrichment efficiency. The design was tested against two benchmark designs, a spiral microfluidic channel and a spiral microfluidic channel followed by a widening channel without the hydrofoils, by processing 5 mL of healthy blood samples spiked with 100 MCF-7 cells. The results proved that the design with hydrofoil-shaped pillars perform significantly better in terms of recovery (recovery rate of 67.9% compared to 23.6% in spiral and 56.7% in spiral with widening section), at a cost of slightly lower white blood cell (WBC) depletion (depletion rate of 94.2% compared to 98.6% in spiral and 94.2% in spiral with widening section), at 1500 µL/min flow rate. For analytical validation, the design was further tested with A549, SKOV-3, and BT-474 cell lines, yielding recovery rates of 62.3 ± 8.4%, 71.0 ± 6.5%, and 82.9 ± 9.9%, respectively. The results are consistent with the size and deformability variation in the respective cell lines, where the increasing size and decreasing deformability affect the recovery rate in a positive manner. The analysis before and after the microfluidic chip process showed that the process does not affect cell viability.

Silibinin Induces Apoptosis and G2/M Cell Cycle Arrest in Human Ovarian Cancer SKOV-3 Cell Line

Background: Silibinin, an herbal polyphenolic flavonolignan, has antioxidant and anticancer properties. Objectives: This study aimed to evaluate some cellular and molecular effects of silibinin on the human ovarian cancer SKOV-3 cell line. Methods: For cytotoxicity investigations of silibinin, MTT assay was used at 24, 48, and 72 hours. Apoptosis and cell cycle were studied by flow cytometry. The effect of silibinin on mRNA expression of B-cell lymphoma 2 (Bcl-2), cyclin E, and S-phase kinase-associated protein 2 (SKP2) was determined by Quantitative reverse transcription polymerase chain reaction (QRT-PCR). Results: Silibinin administration in lower concentrations (12.5 and 25 µg/mL) did not lead to significant (P < 0.05) changes in cell viability and even slightly increased cell growth after 72 hours. However, silibinin in higher concentrations (≥ 50µg/mL) inhibited SKOV-3 cell proliferation in a dose- and time-dependent manner. The mode of cell growth inhibition was apoptosis induction and G2/M cell cycle arrest. Silibinin caused down-regulation of the anti-apoptotic gene, namely Bcl-2. Additionally, silibinin resulted in down-regulation of the major genes in the cell cycle, including cyclin E and SKP2. Conclusions: Overall, this study confirmed the ability of silibinin to suppress ovarian cancer progression through the induction of apoptosis and inhibition of G2/M phase transition. Silibinin may be considered an efficient and safe herbal medication for ovarian cancer.

Structural design, and biological assessment of Co2+, Ni2+, Cu2+and Zn2+ complexes with N-(phenyl)-2-imino-2H-chromene-3-carboxamide

A new series of complexes based on N-(phenyl)-2-imino-2H-chromene-3-carboxamide ligand (NPICC) with Co2+, Ni2+, Cu2+ and Zn2+ metal ions have been synthesized and characterized using IR, H-NMR, UV–Vis, and TGA techniques. In vitro antimicrobial activity of the synthesized compound were conducted using well diffusion method against Gram-positive and Gram-negative strains bacteria and fungi. The results showed that complexes possess good inhibition activity against Klebsiella pneumoniae (G-), staphylococcus epidermis and staphylococcus aureus (G+) as well as candida albicans fungi than ligand and comparable with commercial drugs. Furthermore, the titled compounds were tested for their anti-tumor activity against SKOV-3 and MDA-MB231 cell lines. Results indicated that NiNPICC complex exhibited best activity with IC50 values of 6.42 and 3.95 µg/mL against SKOV-3 and MDA-MB231 cell lines, respectively. To understand the mode of antimicrobial action, quantum mechanics calculation based on DFT theory and docking study were performed for all compounds. Electronic structure analysis and docking energy were investigated. All the titled compounds were docked in a good manner within the active sites of the bacterial and fungi enzymes and exhibited good binding energies. Complexes showed higher electronic activity than ligand supporting the experimental results obtained.

Small Extracellular Vesicles from adipose-derived stem cells suppress cell proliferation by delivering the let-7 family of microRNAs in ovarian cancer

IntroductionOvarian cancer is the leading cause of death among women with gynecological cancer, and novel treatment options are urgently needed. Extracellular vesicles (EVs), including exosomes, may be one of the most promising therapeutic tools for various diseases. In this study, we aimed to investigate the therapeutic effects of adipose-derived stem cell-derived EVs (ADSC-EVs) on ovarian cancer cell lines. Materials and methodsADSCs and the ovarian cancer cell lines SKOV3 and OV90 were used for analysis. ADSC-EVs were isolated through ultracentrifugation and validated using a cryotransmission electron microscope, nanoparticle tracking analysis, and western blotting. Then, the effect of ADSC-EVs on ovarian cancer cells was investigated using IncuCyte and microRNA sequencing. Moreover, the potential functions of miRNAs were evaluated by gain-of function analysis and in silico analysis. ResultsADSC-EVs suppressed SKOV3 and OV90 cell proliferation. In particular, small EVs (sEVs) from ADSCs exhibited a stronger antitumor effect than ADSC-medium/large EVs (m/lEVs). Comparison of the miRNA profiles between ADSC-sEVs and ADSC-m/lEVs, along with downstream pathway analysis, suggested the involvement of the let-7 family. Overexpression of hsa-let-7b-5p and hsa-let-7e-5p significantly suppressed the proliferation of SKOV3 cells. In silico analysis revealed that four potential target genes of hsa-let-7b-5p and hsa-let-7e-5p were significantly associated with the prognoses of the patients. ConclusionADSC-sEVs had a stronger antitumor effect than ADSC-m/lEVs. Hsa-let-7b-5p and hsa-let-7e-5p, which are highly abundant in ADSC-sEVs, suppressed cell proliferation. These findings may open up new possibilities for therapeutic approaches using ADSC-sEVs.

Exosomes derived from ovarian cancer cells regulate proliferation and migration of cancer-associated fibroblasts

Cancer-associated fibroblast (CAF) is an essential risk factor for ovarian cancer. Exosomes can mediate cellular communication in the tumour microenvironment, but the interaction of tumour cell exosomes with CAF is less studied in Ovarian cancer. This study identified H19/miR-29c-3p/LOXL2-COL1A1 as a ceRNA regulatory network involved in regulating tumour matrix-associated signaling pathways associated with CAF. Cellular assays demonstrated that exosomes from ovarian cancer cell line SKOV3 significantly promoted the proliferation and migration of CAF. The results of mixed transplantation tumour experiments in nude mice showed that exosomes of SKOV3 significantly promoted tumour growth. Ovarian cancer tumour-derived exosomes can regulate CAF proliferation and migration through H19/miR-29c-3p/LOXL2-COL1A1. This study reveals the regulatory role of tumour exosomes on CAF, which may provide a theoretical basis for the development of therapeutic regimens targeting fibroblasts in ovarian cancer.

DOCKING STUDIES OF INDOLE ASSIMILATED PYRAZOLINE MOLECULAR HYBRIDS: DESIGN, SYNTHESIS AS ANTIINFLAMMATORY AGENTS AND ANTICANCER AGENTS

A series of novel compounds (4a-4l) have been synthesized by Claisen-Schmidt condensation, Schiff’s base and Cyclization mechanism. All the molecule structures were affirmation by IR, 1H NMR and Mass spectral data. The synthesized hybrids were screened for in vivo anti-inflammatory and invitro anticancer activities. Ant inflammatory activity was performed using carrageenan induced rat paw edema method using Diclofenac as standard drug. The anticancer activity has been assessing by using MTT assay against MCF7 and SKOV3 cell lines and Doxorubicin was used as reference standard. Among the compounds compound 4l exhibited the highest % inhibition of 98.4 when compared with the standard Diclofenac 94.2%.Compound 4f exhibited the lowest IC50 at concentration of 20.01µg against MCF7 cell lines and 32.87µg against SKOV3 cell lines. The molecular docking studies was performed using the Ligprep tool of Schrodinger suite. This study revealed that novel thiazolidne-4-one-pyrazole hybrids(4a-4j) had good interaction with the active site of EGFR receptor. Among the docked compounds, dock score of the compounds ranged from from -3.186 to -5.212. The highest score was exhibited by 4f with -5.212 with Glide binding energy of -34.697 Kcal/mol.

Phytochemical analysis and biological activity of Corchorus olitorius L.: Quantitative analysis of bioactive compounds by LC–MS/MS, antibacterial, enzyme inhibition, and cytotoxic activities

IntroductionCorchorus olitorius L. is a significant plant in folk medicine, therefore, research on phytochemistry with biological activity may reveal its potential use in drug development. Medicinal plants are valuable sources of drug materials as they include bioactive compounds known as secondary metabolites. Determination of bioactive compounds and biological activity of plants lead to understanding their potential in drug development. MethodsA quantitative analysis of the bioactive compounds in leaf, stem, and root extracts of C. olitorius L. was carried out with Liquid Chromatography-Mass Spectrometry (LC–MS/MS). Acetylcholinesterase (AChE) and Glutathione-S-Transferase (GST) enzyme inhibition and antibacterial effects were investigated using Mueller Hinton broth microdilution assay. The anticancer activity of C. olitorius leaves was evaluated by MTT assay using HDF, U87, Skov-3, and Caco-2 cell lines. ResultsIn the quantitative study, leaf extract exhibited the largest concentration of bioactive compounds, and cynarin (6.680 mg/g extract) and chlorogenic acid (5.605 mg/g extract) were the major products. The leaf, stem, and root extracts showed significant AChE and GST inhibition activity. The leaf extract displayed antibacterial activity against P. aeruginosa with the value of 2.2±0.1 (IC50, μg/ mL) in comparison to the standard (4.00±0.1 IC50, μg/ mL). The leaf extract displayed good antiproliferative activity against sk-ov-3 cell lines with values of 37.1% and 42.1% at 250 μg/mL and 500 μg/mL concentrations, respectively. ConclusionC. olitorius has the potential to be a drug agent against cancer and microbial-induced diseases due to its effective bioactive compound contents.

A novel imidazo[1,2-a]pyridine derivative and its co-administration with curcumin exert anti-inflammatory effects by modulating the STAT3/NF-κB/iNOS/COX-2 signaling pathway in breast and ovarian cancer cell lines.

Imidazo[1,2-a]pyridine derivatives with diverse pharmacological properties and curcumin, as a potential natural anti-inflammatory compound, are promising compounds for cancer treatment. This study aimed to synthesize a novel imidazo[1,2-a]pyridine derivative, (MIA), and evaluate its anti-inflammatory activity and effects on nuclear factor-κB (NF-κB) and signal transducer and activator of transcription 3 (STAT3) pathways, and their target genes, alone and in combination with curcumin, in MDA-MB-231 and SKOV3 cell lines. We evaluated the interaction between imidazo[1,2-a]pyridine ligand, curcumin, and NF-κB p50 protein, using molecular docking studies. MTT assay was used to investigate the impacts of compounds on cell viability. To evaluate the NF-κB DNA binding activity and the level of inflammatory cytokines in response to the compounds, ELISA-based methods were performed. In addition, quantitative polymerase chain reaction (qPCR) and western blotting were carried out to analyze the expression of genes and investigate NF-κB and STAT3 signaling pathways. Molecular docking studies showed that MIA docked into the NF-κB p50 subunit, and curcumin augmented its binding. The MTT assay results indicated that MIA and its combination with curcumin reduced cell viability. According to the results of the ELISA-based methods, MIA lowered the levels of inflammatory cytokines and suppressed NF-κB activity. In addition, real-time PCR and Griess test results showed that the expression of cyclooxygenase-2 (COX-2) and inducible NO synthase (iNOS) genes, and nitrite production were reduced by MIA. Furthermore, the western blotting analysis demonstrated that MIA increased the expression of inhibitory κB (IκBα) and B-cell lymphoma 2 (Bcl-2)-associated X proteins (BAX), and suppressed the STAT3 phosphorylation, and Bcl-2 expression. Our findings revealed that curcumin had a potentiating role and enhanced all the anti-inflammatory effects of MIA. This study indicated that the anti-inflammatory activity of MIA is exerted by suppressing the NF-κB and STAT3 signaling pathways in MDA-MB-231 and SKOV3 cancer cell lines.

Biosynthesis, characterization, and investigation of antimicrobial and cytotoxic activities of silver nanoparticles using Solanum tuberosum peel aqueous extract

Metallic nanoparticle biosynthesis is thought to offer opportunities for a wide range of biological uses. The green process of turning biological waste into utilizable products gaining attention due to its economical and eco-friendly approach in recent years. This study reported the ability of Solanum tuberosum (ST) peel extract to the green synthesis of non-toxic, stable, small-sized silver nanoparticles without any toxic reducing agent utilizing the phytochemical components present in its structure. UV–visible spectroscopy, X-ray diffraction analysis, Fourier transform infrared spectroscopy, flourier scanning electron microscopy, atomic force microscopy, transmission electron microscopy, and energy dispersive analysis X-ray confirmed the biosynthesis and characterization of silver nanoparticles. Also, dynamic light scattering and thermogravimetric analyses showed stable synthesized nanoparticles. The antibacterial activity of the biosynthesized silver nanoparticles was evaluated against four different bacterial strains, Escherichia coli (E. coli), Pseudomonas aeruginosa (P. aeruginosa), Staphylococcus aureus (S. aureus) Bacillus subtilis (B. subtilis), and a yeast, Candida albicans (C. albicans) using the minimum inhibitory concentration technique. The cytotoxic activities were determined against Human dermal fibroblast (HDF), glioblastoma (U118), colorectal adenocarcinoma (CaCo-2), and human ovarian (Skov-3) cell lines cancer cells using MTT test. The nanoparticle capping agents that could be involved in the reduction of silver ions to Ag NPs and their stabilization was identified using FTIR. Nanoparticles were spherical in shape and had a size ranging from 3.91 to 27.07 nm, showed crystalline nature, good stability (−31.3 mV), and the presence of capping agents. ST-Ag NPs significantly decreased the growth of bacterial strains after treatment. The in vitro analysis showed that the ST-Ag NPs demonstrated dose-dependent cytotoxicity against cell lines. Based on the data, it is feasible to infer that biogenic Ag NPs were capped with functional groups and demonstrated considerable potential as antibacterial and anticancer agents for biomedical and industrial applications.

Chromosome instability region analysis and identification of the driver genes of the epithelial ovarian cancer cell lines A2780 and SKOV3.

Epithelial ovarian cancer (EOC) is one of the most prevalent gynaecological cancers worldwide. The molecular mechanisms of serous ovarian cancer (SOC) remain unclear and not well understood. SOC cases are primarily diagnosed at the late stage, resulting in a poor prognosis. Advances in molecular biology techniques allow us to obtain a better understanding of precise molecular mechanisms and to identify the chromosome instability region and key driver genes in the carcinogenesis and progression of SOC. Whole-exome sequencing was performed on the normal ovarian cell line IOSE80 and the EOC cell lines SKOV3 and A2780. The single-nucleotide variation burden, distribution, frequency and signature followed the known ovarian mutation profiles, without chromosomal bias. Recurrently mutated ovarian cancer driver genes, including LRP1B, KMT2A, ARID1A, KMT2C and ATRX were also found in two cell lines. The genome distribution of copy number alterations was found by copy number variation (CNV) analysis, including amplification of 17q12 and 4p16.1 and deletion of 10q23.33. The CNVs of MED1, GRB7 and MIEN1 located at 17q12 were found to be correlated with the overall survival of SOC patients (MED1: p = 0.028, GRB7: p = 0.0048, MIEN1: p = 0.0051), and the expression of the three driver genes in the ovarian cell line IOSE80 and EOC cell lines SKOV3 and A2780 was confirmed by western blot and cell immunohistochemistry.

Probing supramolecular complexation of the drug benserazide hydrochloride with hydroxypropyl-β-cyclodextrin by experimental and computational studies

Herein, we explore the supramolecular complexation of Benserazide hydrochloride (BNZ) with Hydroxypropyl-β-cyclodextrin (HP-β-CD) in an aqueous medium by means of UV–visible, surface tension, FT-IR, 1H NMR, 2D ROESY, DSC, PXRD and molecular docking study. These studies suggest the effective complexation of BNZ with HP-β-CD resulting in the formation of BNZ-HP-β-CD complex. Surface tension study and Job's plot confirm 1:1 stoichiometry of BNZ-HP-β-CD complex. FT-IR, 1H NMR and 2D ROESY studies suggest the possible binding mode of BNZ into the cavity of HP-β-CD. The binding constant (Ka) of BNZ-HP-β-CD complex indicates the affinity of HP-β-CD for BNZ. The estimated negative free energy of binding reveals that the complexation process is thermodynamically feasible. PXRD analysis confirms the formation of BNZ-HP-β-CD inclusion complex. DSC analysis shows the enhancement in the thermal stability of BNZ after complexation with HP-β-CD. The molecular docking study introduces the most stable binding orientation of BNZ within the cavity of HP-β-CD. The complex expresses better cytotoxic effect than free BNZ on both lung carcinomic A549 cell line and ovarian SKOV3 cancer cell line. Furthermore, the considerable improvement in the antioxidant activity of BNZ is registered after encapsulation.

Tasquinimod enhances the sensitivity of ovarian cancer cells to cisplatin by regulating the Nur77-Bcl-2 apoptotic pathway.

Resistance to cisplatin (DDP) in ovarian cancer therapy has been a major clinical barrier. Drug-resistant cancers have been shown to downregulate the proapoptotic protein B-cell lymphoma-2 (Bcl-2) to inhibit apoptosis. Therefore, we explored whether tasquinimod could modulate resistance to DDP through apoptotic pathways. We aimed to explore the relationship between tasquinimod, Nur77-Bcl-2 apoptosis pathway and sensitivity of the ovarian carcinoma cell line SKOV3 and the DDP-resistant strain SKOV3/DDP cells to DDP. First, SKOV3 and SKOV3/DDP cells were treated with 2 μg/mL DDP or 40 μM tasquinimod. Western blot and quantitative real-time polymerase chain reaction (qPCR) were then used to analyze the expression of histone deacetylase 4 (HDAC4), Nur77, Bcl-2 (BH3 domain-specific), and caspase-3. Flow cytometry, scratch-wound assay and immunofluorescence were used to detect apoptosis, migration rate, and related expression of Nur77 and Bcl-2 (BH3 domain-specific). Subsequently, 5×107 SKOV3 or SKOV3/DDP cells cultured with 2 μg/mL DDP were injected into 4-week-old female BALB/c nude mice. Then, the mice were administered 4 mg/kg DDP and 50 mg/kg tasquinimod every 3 days. Finally, the changes in tumor diameter and weight were measured. After treatment of SKOV3 and SKOV3/DDP cells with tasquinimod, cell migration and HDAC4 expression levels were significantly reduced, while Nur77 expression was increased. Tasquinimod treatment enhanced the expression of Nur77 and caspase-3, and cells transfected with si-Nur77 showed the opposite result. Transfection of si-Nur77 reduced the expression of caspase-3 and Nur77 in the SKOV3/DDP cells that were treated with both DDP and tasquinimod. After injection of SKOV3/DDP cells into the mice, the tumor diameter, mass and in vivo HDAC4 level were significantly decreased by tasquinimod. Meanwhile, the levels of Nur77 and Bcl-2 (BH3 domain-specific) were increased. Tasquinimod upregulated the Nur77/Bcl-2 pathway to induce apoptosis in SKOV3/DDP cells and enhanced the anti-tumor effect of DDP in SKOV3/DDP xenografts. Therefore, tasquinimod can be expected to find clinical applications in enhancing DDP resistance.

Novel Iron Chelator SK4 Drives Cytotoxicity through Inhibiting Mitochondrial Metabolism in Ovarian and Triple Negative Breast Cancer Cell Lines.

Anti-cancer therapy by iron chelation has been shown to inhibit many cellular processes including DNA replication, mitochondrial metabolism and oncogenic signalling pathways (e.g., EGFR). Iron chelator SK4 represents a double pronged approach towards treating cancer. SK4 enters through LAT1, a commonly overexpressed amino acid transporter in tumours, thus targeting iron addiction and LAT1 overexpression. The aim of this study was to characterise the mode of action of SK4 through proteomics, metabolomics, lipidomics and seahorse real-time analysis in ovarian cell line SKOV3 and triple negative breast cancer cell line MDA MB 231. Pathway enrichment of proteomics data showed an overrepresentation of metabolism related pathways. Metabolic change after SK4 exposure have been confirmed in investigations of changes in basal and maximal mitochondrial respiration using seahorse real-time analysis of mitochondrial metabolism. Metabolomics also showed an increase in AMP and glucose-1-phosphate. Interestingly, our lipidomics data show a decrease in phospholipid synthesis in the SKOV3 cells which is in contrast with previous data which showed an upregulation of ceramide driven apoptosis. In summary, our data highlight impairment of energy metabolism as a mechanism of action underlying SK4 apoptosis, but also suggest a potential role of ceramide induction in the phenotypic outcome of the cell model.

Machine learning and biological evaluation-based identification of a potential MMP-9 inhibitor, effective against ovarian cancer cells SKOV3

MMP-9, also known as gelatinase B, is a zinc-metalloproteinase family protein that plays a key role in the degradation of the extracellular matrix (ECM). The normal function of MMP-9 includes the breakdown of ECM, a process that aids in normal physiological processes such as embryonic development, angiogenesis, etc. Interruptions in these processes due to the over-expression or downregulation of MMP-9 are reported to cause some pathological conditions like neurodegenerative diseases and cancer. In the present study, an integrated approach for ML-based virtual screening of the Maybridge library was carried out and their biological activity was tested in an attempt to identify novel small molecule scaffolds that can inhibit the activity of MMP-9. The top hits were identified and selected for target-based activity against MMP-9 protein using the kit (Biovision K844). Further, MTT assay was performed in various cancer cell lines such as breast (MCF-7, MDA-MB-231), colorectal (HCT119, DL-D-1), cervical (HeLa), lung (A549) and ovarian cancer (SKOV3). Interestingly, one compound viz., RJF02215 exhibited anti-cancer activity selectively in SKOV3. Wound healing assay and colony formation assay performed on SKOV3 cell line in the presence of RJF02215 confirmed that the compound had a significant inhibitory effect on this cell line. Thus, we have identified a novel molecule that can inhibit MMP-9 activity in vitro and inhibits the proliferation of SKOV3 cells. Novel molecules based on the structure of RJF02215 may become a good value addition for the treatment of ovarian cancer by exhibiting selective MMP-9 activity. Communicated by Ramaswamy H. Sarma

Development of a Radiolabeled Folate-Mediated Drug Delivery System for Effective Delivery of Docetaxel.

Many preclinical studies are carried out with the aim of developing new formulations for the effective delivery of taxane class drugs, one of the most important anticancer drugs used clinically today. In this study, a radiolabeled folate-mediated solid lipid magnetic nanoparticle (SLMNP) system was developed by loading superparamagnetic iron oxide nanoparticles (MNP) and docetaxel (DTX) into the solid lipid nanoparticles as a drug delivery system that will function both in cancer treatment and diagnosis. For this purpose, first, SLMNP was synthesized by the hot homogenization method, and the surface of the particles was modified with a folate derivative to carry the particles to tissues with folate receptors. The synthesized magnetic solid lipid nanoparticles were loaded with DTX, and then radiolabeling was carried out with technetium-99 m (99mTc-DTX-SLMNP). Structural characteristics of these nanoparticles were determined by characterization methods. According to the TEM images of MNPs, SLN, and SLMNPs, MNPs were observed between 25and 35 nm, SLNs between 400 and 500 nm, and SLMNPs between 350 and 450 nm. The drug entrapment efficiency of SLMNPs loaded with DTX was found to be 19%, and the percentage efficiency of radiolabeling was found to be 98.0 ± 2.0%. The biological behavior of this radiolabeled system was investigated in vitro and in vivo. Folate receptor-positive SKOV-3 and folate receptor-negative A549 cancer cell lines were studied. The IC50 values of DTX-SLMNP in SKOV-3 and A549 cells were 50.21 and 172.27 μM at 48 h, respectively. Gamma camera imaging studies of 99mTc-DTX-SLMNP and magnetically applied 99mTc-DTX-SLMNP compounds were performed on tumor-bearing CD-1 nude mice. The uptake in the folate receptor-positive tumor region was higher than that in the folate receptor negative tumor region. We proposed that the drug delivery system we prepared in this study be evaluated for preclinical studies of new drug carrier formulations of the taxane class of anticancer drugs.

Ginsenoside 20(S)-Rg3 reduces KIF20A expression and promotes CDC25A proteasomal degradation in epithelial ovarian cancer

BackgroundGinsenoside 20(S)-Rg3 shows promising tumor-suppressive effects in ovarian cancer via inhibiting NF-κB signaling. This study aimed to explore the downstream tumor suppressive mechanisms of ginsenoside Rg3 via this signaling pathway. Materials and methodsA systematical screening was applied to examine the expression profile of 41 kinesin family member genes in ovarian cancer. The regulatory effect of ginsenoside Rg3 on KIF20A expression was studied. In addition, we explored interacting proteins of KIF20A and their molecular regulations in ovarian cancer. RNA-seq data from The Cancer Genome Atlas (TCGA) was used for bioinformatic analysis. Epithelial ovarian cancer cell lines SKOV3 and A2780 were used as in vitro and in vivo cell models. Commercial human ovarian cancer tissue arrays were used for immunohistochemistry staining. ResultsKIF20A is a biomarker of poor prognosis among the kinesin genes. It promotes ovarian cancer cell growth in vitro and in vivo. Ginsenoside Rg3 can suppress the transcription of KIF20A. GST pull-down and co-immunoprecipitation (IP) assays confirmed that KIF20A physically interacts with BTRC (β-TrCP1), a substrate recognition subunit for SCFβ−TrCP E3 ubiquitin ligase. In vitro ubiquitination and cycloheximide (CHX) chase assays showed that via interacting with BTRC, KIF20A reduces BTRC-mediated CDC25A poly-ubiquitination and enhances its stability. Ginsenoside Rg3 treatment partly abrogates KIF20A overexpression-induced CDC25A upregulation. ConclusionThis study revealed a novel anti-tumor mechanism of ginsenoside Rg3. It can inhibit KIF20A transcription and promote CDC25A proteasomal degradation in epithelial ovarian cancer.

Synthesis, characterization, and anticancer properties of iminopyridine platinum(II) complexes containing long aliphatic chains

A series of iminopyridine ligands were prepared from the condensation reaction of 6-methylpyridine-2-carboxaldehyde and long-chain aliphatic primary amines. Square planar platinum(II) complexes were synthesized from the reaction of [PtCl2(η2-coe)]2 (coe = cis-cyclooctene) with two equivalents of ligand. The synthesis of the platinum compounds appears to go through a five-coordinate trigonal pyramidal species where coe remains coordinated initially but eventually dissociates to afford the four-coordinate complexes. Ligands and platinum complexes were characterized using multi-nuclear NMR and FT-IR spectroscopies as well as an X-ray diffraction study for the platinum complex derived from n-hexylamine. The cytotoxic properties of the platinum complexes against the ovarian cancer cell line SKOV-3 using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide colorimetric method were examined. Relative cytotoxicities of the platinum complexes follow a trend of greater potency with increasing chain length against the SKOV-3 cell line, with the longest aliphatic hydrocarbon chain (C16) exhibiting anticancer effects comparable to that of cisplatin.