Cervical Cell Carcinoma Research Articles

Peptide direct growth on poly(acrylic acid)/poly(vinyl alcohol) electrospun fibers coated with branched poly(ethylenimine): A solid-phase approach for scaffolds biofunctionalization

Due to their resemblance to the fibrillar structure of the extracellular matrix, electrospun nanofibrous meshes are currently used as porous and mechanically stable scaffolds for cell culture. In this study, we propose an innovative methodology for growing peptide sequences directly onto the surface of electrospun nanofibers. To achieve this, electrospun fibers were produced from a poly(acrylic acid)/poly(vinyl alcohol) blend that was thermally crosslinked and subjected to a covalent coating of branched poly(ethylenimine). The exposed amino functionalities on the fiber surface were then used for the direct solid-phase synthesis of the RGD peptide sequence. In contrast to established strategies, mainly involving the grafting of pre-synthesized peptides onto the polymer chains before electrospinning or onto the nanofibers surface, this method allows for the concurrent synthesis and anchoring of peptides to the substrate, with potential applications in combinatorial chemistry. The incorporation of this integrin-binding motive significantly enhanced the nanofibers' ability to capture human cervical carcinoma (HeLa) cells, selected as a proof of concept to assess the functionalities of the developed material.

Ethanolic extract from Sophora moorcroftiana inhibit cell proliferation and alter the mechanical properties of human cervical cancer

BackgroundCervical cancer is one of the most common gynecological malignancies. Previous studies have shown that the ethanol extract of Sophora moorcroftiana seeds (EESMS) possesses an antiproliferative effect on several tumors in vitro. Therefore, in this study, we assessed the impact of EESMS on human cervical carcinoma (HeLa) cell proliferation.MethodsThe proliferation and apoptotic effects of HeLa cells treated with EESMS were evaluated using 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide assay, dual acridine orange/ethidium bromide double staining, flow cytometry, and western blotting. Single-cell level atomic force microscopy (AFM) was conducted to detect the mechanical properties of HeLa cells, and proteomics and bioinformatics methods were used to elucidate the molecular mechanisms of EESMS.ResultsEESMS treatment inhibited HeLa cell proliferation by blocking the G0/G1 phase, increasing the expression of Caspase-3 and affecting its mechanical properties, and the EESMS indicated no significant inhibitory effect on mouse fibroblasts L929 cell line. In total, 218 differentially expressed proteins were identified using two-dimensional electrophoresis, and eight differentially expressed proteins were successfully identified using matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry. The differentially expressed proteins were involved in various cellular and biological processes.ConclusionThis study provides a perspective on how cells change through biomechanics and a further theoretical foundation for the future application of Sophora moorcroftiana as a novel low-toxicity chemotherapy medication for treating human cervical cancer.

A plasma-based proteomic platform for predicting clinical benefit from immune checkpoint inhibitors in multiple cancers.

2568 Background: Immune checkpoint inhibitor (ICI)-based therapies are preferred treatment options for multiple cancer types. However, there is an unmet need for predictive tests that can identify patients likely to benefit. PROphet-NSCLC is a commercially available pretreatment plasma proteomic test that predicts clinical benefit from first-line PD-(L)1 inhibitor-based therapy in patients with metastatic non-small cell lung cancer (NSCLC), guiding the choice between ICI monotherapy versus ICI-chemotherapy. Here, we evaluate the broader clinical utility of PROphet-NSCLC by testing its predictive capabilities across diverse cancer types. Methods: Pre-treatment plasma samples and clinical data were collected for an observational study from patients with metastatic melanoma (n=68) and HPV-related cancers (n=43; NCT03427411) treated with PD-(L)1 inhibitor-based therapies. HPV-related cancers included anogenital squamous cell carcinoma (n=16), cervical carcinoma (n=18), and head and neck squamous cell carcinoma (n=9). Patients were assigned a PROphet-positive or -negative result based on computational analysis of SomaScan-derived proteomic profiles in pre-treatment plasma samples. Overall survival (OS) and progression-free survival (PFS) in PROphet-positive- and -negative groups was analyzed with the Kaplan-Meier method. Hazard ratios (HR) were calculated from univariate and multivariate Cox proportional hazard models. Results: In the melanoma cohort, PROphet-positive patients (n=51) displayed a significant OS benefit in comparison to PROphet-negative patients (n=17; median OS 92.8 months vs. 9.5 months, HR=0.14, 95% CI: 0.06-0.34, p<0.0001), consistent with the clinically validated predictive performance of the test in patients with NSCLC. OS results remained significant after correcting for sex, age, histology, ECOG performance status, and treatment type (HR=0.05, 95% CI: 0.01-0.25, p<0.001). In PROphet-positive patients, PD-1+CTLA-4 inhibitor combination therapy (n=27) was superior to PD-1 inhibitor monotherapy (n=24; OS: median 118.4 vs. 48.3 months, HR=0.58, p=0.24; PFS: median not reached vs. 10.8 months, HR=0.48, p=0.04). PROphet-negative patients displayed similarly poor outcomes with either treatment, providing a rationale to consider alternative therapies for such patients. In the HPV-related cohort, the median OS in PROphet-positive (n=10) vs PROphet-negative (n=33) groups was 43.6 vs 4.4 months (HR=0.22, 95% CI: 0.08-0.59, p=0.001), with similar trends per sub-cohort. Conclusions: Our findings show that the PROphet-NSCLC test can be applied to PD-(L)1 inhibitor-treated melanoma and HPV-related cancers, suggesting applicability to cancers beyond NSCLC. Analysis of additional patient samples is needed to explore the potential utility of PROphet-NSCLC for informing treatment decisions for a broad range of cancer types.

Isolation of flavonoids from Tagetes patula(French Marigold) flowers: Cytotoxic and oxidant activity in human cervical carcinoma cells

Tagetes patula, known as French Marigold, belongs to the family Asteraceae. Human papillomavirus infection is considered one of the causes of cervical cancer. This study assessedthe cytotoxic activity and intracellular oxidative capacity of compounds isolated from extract of T. patulaflowers as anti-cancer cervical agents. Fraction F6 of n-butanol extract was subjected to column chromatography and HPLC-ESI-MS. The isolated compounds of T. patulawere used to examine cytotoxic activity and the production of total reactive oxygen species in SiHa and HeLa cells; the cells were also characterized using scanning electron microscopy. Patulitrin was cytotoxic to SiHa and HeLa cells. An increase in ROS production was observed at different times of treatment of cells with patuletin and patulitrin. Scanning electron microscopy showed morphological changes in SiHa and HeLa cells. Thus, compounds isolated from T. patulahave great treatment potential against cervical cancer.

Combined Effects of Annexin A5 Overexpression, 5-Fluorouracil Treatment, and Irradiation on Cell Viability of Caski Cervical Cancer Cell Line.

Cervical cancer is the fourth leading cause of cancer deaths in women globally. Combining gene therapy with chemo- and radiotherapy may improve cervical cancer treatment outcomes. This study evaluated the effects of Annexin A5(ANXA5) overexpression alongside 5-fluorouracil (5-FU) and irradiation on the viability of CaSki cervical squamous cell carcinoma (SCC) cells. pAdenoVator-CMV-ANXA5-IRES-GFP-plasmid and mock plasmid were transfected into CaSki cells using calcium-phosphate. Seventy-two hours post-transfection, GFP expression was quantified by fluorescence microscopy and flow cytometry to evaluate transfection efficiency. ANXA5 overexpression was confirmed via qPCR. Twenty-four hours post-transfection, cells received a single dose of 8Gy and were treated with 1 and 2µg/ml of 5-FU (IC50 = 2.783µg/ml). Cell viability, apoptosis, cell cycle stage, and Bcl-2 and Bax gene expression were assessed via MTT, annexin V/7-AAD, PI staining, and qPCR assays, respectively. ANXA5 was overexpressed 31.5-fold compared to control (p < 0.0001). MTT assays showed ANXA5 overexpression dose-dependently reduced CaSki cell viability (p < 0.001). IC50 of 5-FU was reduced from 2.783μg/mL to 1.794μg/mL when combined with ANXA5 overexpression. Additive effects on cell death were observed for ANXA5 plus 5-FU or irradiation versus ANXA5 alone. Apoptosis assays indicated combinatorial treatment increased CaSki cell apoptosis over ANXA5 alone. Cell cycle analysis revealed ANXA5 arrested cell cycle at G1/S phases; the percentage of cells in the S phase further rose with combination treatment. Finally, combination therapy significantly decreased Bcl-2 expression and increased Bax versus control (p < 0.001). Altogether, ANXA5 overexpression alongside 5-FU and irradiation may improve cervical squamous cell carcinoma (SCC) treatment efficacy. Further, in vivo investigations are warranted to confirm these in vitro results.

Harnessing THz Technology: Biosensor for Highly Accurate Cervical Cancer Cell Detection via Refractive Index.

In order to rapidly identify various species of cancer cells in the tissues of person, a unique diamond shaped hollow-core photonic crystal fiber (PCF)-formed by optical waveform is developed and computationally studied. In this investigation, we found the most prevalent cancers, such as HeLa-derived cervical carcinoma. Since normal and cancer cells differ in their refractive indices (RIs), other significant optical properties can be assessed using this information. With the use of the finite element method, a computational tool for solving simultaneous equations, the defining characteristics the suggested cancer cell sensor are examined using COMSOL-Multiphysics software. Additionally, strict mesh parts are used to preserve the utmost level of modeling realism. At 2.4 THz, the PCF detector attains a Relative Sensitivity of around 97.51% and 96.29%, Confinement Loss of 6.1 × 10 -09db/m and 4.39 × 10-07db/m with respect to cervical carcinoma cell and cervical normal cell. The straightforward PCF structure provides a wide chance of application using the continuing fabrication technique, based on these conventional values of performance indices. This biosensor utilizes the distinctive refractive characteristics of cancer cells, providing a highly accurate and dependable approach for the early identification of cervical cancer. This has the potential to significantly transform the process of cervical cancer screening. The novel method boosts the ability to detect and identify certain conditions, leading to increased diagnostic capabilities for early treatment and better results for patients.

Dissimilar effect of organometallic ruthenium complexes on the viability of MDR and non-MDR experimental models

Ruthenium complexes containing triphenylphosphine diamide ligands were prepared, characterized, and tested for their biological activity against various cancer cell lines and the malaria parasite, Plasmodium falciparum. The effect of M (mono-substituted) and B (bis-substituted) complexes on the human cervical carcinoma (HeLa) cell line was investigated using the MTT assay. Five (B2, B3, B5, B6, and B13) of the 24 synthesized ruthenium complexes showed significant effects with IC50 values ranging between 0.3 and 2.3 μM. Evaluation of the potential biomolecular targets of B2 and B13 by fluorescence spectroscopy revealed relevant interactions with BSA and only a weak affinity for ctDNA. Complexes M2, B2, M13 and B13 were selected for further biological characterization. Their effect on the viability of two ovarian cancer cell lines was compared to normal cell lines, denoting their selectivity. Upon treatment of four different drug-resistant gynaecological cancer cell lines, differing in their multidrug-resistant phenotypes, the efficacy of the bis-substituted complexes was shown to be greater than their mono-substituted counterparts. The non-MDR cells are sensitive to all the tested complexes, compared to MDR cells which are less sensitive. Upon investigation of complexes M2, M13, B2, and B13 against sensitive and multidrug-resistant parasite strains of P. falciparum, the bis-substituted complexes were again shown to be the most potent, with submicromolar activity against both strains. Furthermore, the resistance indexes for the complexes were approximately equal to 1, which is at least 5-fold lower than chloroquine diphosphate, suggesting the ability of these complexes to retain their activity in resistant forms of the parasite.

An automated approach for hemocytometer cell counting based on image-processing method

Evaluating cell viability plays an important role in routine cell culture, toxicology, and drug development. Trypan blue stain is one of the simplest assays that are used to determine the number of viable cells in a cell suspension. The stained cell suspension can then be counted under a microscope using a hemocytometer. Hemocytometers have a fixed volume, making it possible to calculate the concentration of cells in the sample and measure viability. Most of the related studies are conducted using manual counting methods. However, manual counting is time-consuming and laborious. While various automated cell counting devices have been proposed, they tend to be too expensive for widespread deployment. Although low-cost automated cell counting methods presented in the literatures provide high accuracy and cost-effective alternatives, only cells in the field of view of microscope with unknown volume are counted. This will make the concentration inestimable. Accordingly, the present study proposes a low-cost but accurate method for hemocytometer cell counting using an image-processing approach. The accuracy of the proposed method is demonstrated by comparing the cell counting results obtained for human cervical carcinoma (HeLa) cells with the manual counts. It is shown that the proposed approach has a mean absolute percentage error of less than 6.26 % for samples containing a mixture of live and dead cells. Moreover, the proposed system is equipped with a simple and intuitive human machine interface such that the counting task can be performed by individuals with no prior experience in the image-processing field.

Glycyrrhizin Impairs the Progression and Metastasis of Cervical Cancer Through the Protein Kinase B/Glycogen Synthase Kinase-3β/β-Catenin Signaling Pathway

The present study assessed the impact of glycyrrhizin on the mechanisms of progression and metastasis in cervical cancer cells. To this end, cell proliferation, migration, and invasion and protein/mRNA expression levels of protein kinase B, glycogen synthase kinase-3 β and β-catenin were measured in three groups of cells: normal cervical epithelial cell line End1/E6E7 (Group A), human cervical squamous cell carcinoma cell line C33a (group B), and human cervical squamous cell carcinoma cell line C33a treated with 50μmol/L glycyrrhizin (Group C). Results showed that glycyrrhizin treatment decreased the proliferation, migration, and invasion of C33a cells and increased apoptosis compared to the cervical cancer control cells. Expression levels of proteins associated with apoptosis such as FS-7-associated surface antigen, Bcl-2-associated X protein and Fas ligand were higher, while protein kinase B pathway activity was attenuated in the glycyrrhizin-treated group compared to the cervical cancer control group. These results suggest that glycyrrhizin may impair cervical cancer cell viability and impede their progression and metastasis by modulating the protein kinase B/glycogen synthase kinase-3β/β-catenin signaling pathway.

The Micro-Immunotherapy Medicine 2LPAPI® Displays Immune-Modulatory Effects in a Model of Human Papillomavirus Type-16 L1-Protein Capsid-Treated Human Peripheral Blood Mononuclear Cells and Antiproliferative Effects in a Model of Cervical Cancer Cells.

Human papillomavirus (HPV) is the second most common infectious agent causing cancer. Persistent infection with high-risk (HR)-HPV can lead to cervical intra-epithelial neoplasia and cervical carcinomas (CC). While host immune response is necessary for viral clearance, chronic immune activation contributes to a low-grade inflammation that can ultimately lead to carcinogenesis. The micro-immunotherapy medicine (MIM) 2LPAPI® could be a valuable tool to manage the clearance of the virus and reduce the risk of developing CC. In this in vitro study, we aimed to investigate its mode of action. We showed that actives from the MIM increased the IL-6, IFN-γ, and IP-10 secretion in human peripheral blood mononuclear cells (PBMCs) exposed to peptides derived from the HPV-16 capsid (HPV16(L1)). This could reflect an increase in the immune activity toward HPV-16. At the same time, some active substances reduced the lympho-proliferation and the expression of T-cell activation markers. Finally, some of the MIM actives displayed antiproliferative effects in CC-derived HeLa cells under serum-starvation conditions. Altogether, this body of data highlighted for the first time the dual effect of MIM in the framework of HR-HPV infections as a potential (i) immune modulator of HPV16(L1)-treated PBMCs and (ii) antiproliferative agent of HPV-positive CC cells.

Parabens promotes invasive properties of multiple human cells: A potential cancer-associated adverse outcome pathway

Parabens are esters of p-hydroxybenzoic acid, which have been used as preservatives and considered safe for nearly a century, until the last two decades when concerns began to be raised about their association with cancers. Knowledge of the mode of action of parabens on the metastatic properties of different cancer cells is still very limited. In the present study, we investigated the effects of methylparaben (MP) and propylparaben (PP) on cell invasion and/or migration in multiple human cancerous and noncancerous cells, including hepatocellular carcinoma cells (HepG2), cervical carcinoma cells (HeLa), breast carcinoma cells (MCF-7), and human placental trophoblasts (HTR-8/SVneo). MP and PP at concentrations in a range of 5–500 μg/L significantly promoted the invasion of four cell lines, with a minimum effective concentration of 5 μg/L. MP and PP up-regulated the expression levels and enzymatic activities of matrix metalloproteinase 2 and 9 (MMP2 and MMP9), as well as altered the expression of the tissue inhibitors of metalloproteinase 1 and 2 (TIMP1 and TIMP2) in four cell lines, suggesting MMPs/TIMPs as potential key events (KEs) for paraben-induced cell invasion. Activation of the p38 mitogen-activated protein kinase (p38 MAPK) and c-Jun N-terminal protein kinases 1/2 (JNK1/2) signaling pathways was required for MP- and PP-promoted invasion of four cell lines, suggesting MAPK signaling pathways as candidates for KEs in cancer or noncancerous cells response to paraben exposure. This study showed for the first time that the two widely used parabens, MP and PP, promoted invasive capacity of multiple human cells through a common mode of action. This study provides evidence for the establishment of a potential cancer-associated AOP for parabens based on pathway-specific mechanism(s), which contributes towards assessing the health risks of these environmental chemicals.

The cytotoxicity of palladium (II) complexes containing 1,2‐ or 1,4‐diazine bridging ligands on squamous cell carcinoma cells in vitro: Insights in the mechanisms of action

Binuclear [{Pd (dien)}2(μ‐pz)](ClO4)4 (Pd1) and [{Pd (dien)}2(μ‐pydz)](ClO4)4 (Pd2) complexes, where pz is 1,4‐diazine and pydz is 1,2‐diazine bridging ligands and dien (diethylenetriamine) is a tridentately coordinated triamine ligand, were synthesized and characterized using elemental analysis, NMR (1H and 13C) spectroscopy, and UV–Vis spectrophotometry. Electronic absorption spectrophotometry and fluorescence spectroscopy were used to study the interactions of the prepared Pd1 and Pd2 complexes with deoxyribonucleic acid (CT‐DNA) and bovine serum albumin (BSA). According to the UV–Vis spectrophotometric data, the complexes interact with CT‐DNA and bind to the BSA as a possible carrier to the target site. The antitumor properties of the tested palladium compounds were evaluated on three types of human cancer cells, namely, squamous cell carcinoma cells SCC‐4, cervical carcinoma cells HeLa and colorectal carcinoma cells HCT116, and one type of malignantly unchanged cells, MRC‐5 fibroblasts. The highest cytotoxicity was shown by the Pd1 compound against SCC‐4 cell, with an IC50 value of 5.4 ± 1.2 μM. The results showed that the cytotoxic potential against SCC‐4 cells of our new palladium complexes Pd1 and Pd2 was comparable with the cytotoxic potential of cisplatin and oxaliplatin. However, Pd1 and Pd2 displayed significantly lower cytotoxicity in comparison with cisplatin and oxaliplatin against non‐malignant cells, fibroblasts MRC5, so these results may suggest that local and systemic complications of antitumor therapy could be reduced and/or avoided. Both compounds selectively decreased the viability of SCC‐4 cells by induction of apoptosis. Also, Pd1 and Pd2 induced S‐phase cell cycle arrest in human SCC‐4 cells, so that could be another mechanism to halt the uncontrolled growth and division of cancer cells. Moreover, the precise molecular mechanism of Pd1 and Pd2 complexes‐induced apoptosis was the activation of the intrinsic signaling pathway of apoptosis, which also included activation of caspase‐3. Therefore, our new palladium (II) complexes might have a promising role as the potential future cytostatic agents.

Plumbagin as a preferential lead molecule to combat EGFR-driven matrix abundance and migration of cervical carcinoma cells.

The handshake between the complex networks of matrix components in the tumor micro-environment (TME) is considered as a crucial event in the progression of several cancers including cervical carcinoma (CC). A number of studies report a connection between epidermal growth factor (EGF) and matrix component production. Studies demonstrate that the mechano-transduction trigger by collagen, influences the tumor cells to undergo epithelial-mesenchymal transition (EMT) and block the entryof drugs. We hypothesize that the intervention to prevent EGF triggered deposition of matrix components could sensitize several therapies for CC cells. We utilized morphological assessment, MTT assay, mitored tracking, acridine orange (AO)/ ethidium bromide (EtBr) staining and bromodeoxyuridine (BrdU) assay to measure the cell viability, mitochondrial activity, cellular apoptosis, and DNA synthesis. Clonogenic assay and scratch healing assay were executed to address the stemness and migratory potential. Detection of glycosaminoglycan's (GAGs), collagen, matrix metalloproteinase (MMP)-2/9 secretion and calcium (Ca2+) ions were performed to assess the production of matrix components. Finally, the interaction between EGFR and plumbagin was evaluated by employing molecular dynamics (MD) simulation. Pre-treating the cells with plumbagin inhibited the EGF-induced EMT along with reduction in cell proliferation, migration, clonogenesis and depletion of matrix components. The actions of EGF and plumbagin were more pronounced in HPV-positive CC cells than HPV-negative CC cells. This study identified that increased matrix production triggered by EGF-rich milieu is inhibited by plumbagin in human papilloma viral (HPV) 68 positive ME180, HPV 16 positive SiHa and HPV-negative C33A cell lines. Delivery of plumbagin directly to TME would effectively accelerate the clearance of CC cells, reduce metastasis and matrix abundance by employing targeted delivery to minimize the undesired effects of plumbagin.

Abstract 1830: Bergenin inhibits growth of human cervical cancer cells by decreasing galectin-3 and MMP-9 expression

Abstract Cervical cancer is still the leading of cancer mortality worldwide, even after introducing a vaccine against Human papillomavirus (HPV), due to low vaccine coverage, especially in the developing world. Cervical cancer is primarily treated by Chemo/Radiotherapy, depending on the disease stage, with Carboplatin/Cisplatin-based drug regime. These drugs being non-specific, target rapidly dividing cells, including normal cells, so safer options are needed for more targeted therapy and lower off-target toxicity. Natural products offer an attractive option compared to synthetic drugs due to their well-established safety profile and capacity to target multiple oncogenic hallmarks of cancer like inflammation, angiogenesis, etc. In the current study, we investigated the effect of Bergenin (C-glycoside of 4-O-methylgallic acid), a natural polyphenol compound that is isolated from medicinal plants such as Bergenia crassifolia, Caesalpinia digyna, and Flueggea leucopyrus. Bergenin has been shown to have anti-inflammatory, anti-ulcerogenic, and wound healing properties. Still, its anticancer potential has been realized only recently. We performed a proteomic analysis of cervical carcinoma cells treated with bergenin and found it to influence multiple hallmarks of cancers, including apoptosis, angiogenesis, and tumor suppressor proteins. It was also involved in many different cellular processes unrelated to cancer, as shown by our proteomic analysis. Further analysis showed bergenin to be a potent -angiogenic agent by reducing key angiogenic proteins like Galectin 3 and MMP-9 (Matrix Metalloprotease 9) in cervical carcinoma cells at cellular level as shown by western blotting assay. Further understanding of this interaction was carried out using molecular docking analysis, which indicated MMP-9 has more affinity for Bergenin as compared to Galectin-3. Cumulatively, our data provide novel insight into the anti-angiogenic mechanism of Bergenin in cervical carcinoma cells by modulation of multiple angiogenic proteins like Galectin-3 and MMP-9 which warrant its further development as an anticancer agent in cervical cancer. Citation Format: Mayank Singh, Ravi Chauhan, Lakshay Malhotra, Ashna Gupta, Gunjan Dagar, Dayasagar Das, Shahab Uddin, Ethayathulla Abdul Samath, Muzafar A. Macha, Ammira Al shabeeb Akil, Ajaz A. Bhat. Bergenin inhibits growth of human cervical cancer cells by decreasing galectin-3 and MMP-9 expression [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 1830.

Single extract of Forsythia Suspense versus the prepared drug in pieces: comparison of their anti-inflammatory, antitumor and antibacterial effects in zebrafish

To compare the anti-inflammatory, antitumor and anti-bacterial effects of the single extract (in granules) and the prepared drug in pieces of Forsythia Suspense (Lianqiao, a traditional Chinese herbal medicine). In zebrafish embryo models of CuSO4 exposure, tail transection and LPS microinjection-induced inflammation, the anti-inflammatory effects of 10 μg/mL DEX, single extract of Forsythia Suspense, and the water extract of the prepared drug (400, 600, and 800 μg/mL) were evaluated by observing neutrophil counts, RT- qPCR, HE staining and survival analysis. Zebrafish embryo models bearing different human tumor cell xenografts were used to assess the anti-tumor effect of the drugs in different dosage forms by fluorescence staining and HE staining. The microbroth dilution method was used to evaluate the antibacterial efficacy of the drugs. In the zebrafish embryo models of inflammation, both of the two dosage forms of Forsythia Suspense significantly inhibited neutrophil aggregation, reduced the mRNA expressions of TNF-α, IL-6, P38, Jnk, Erk and P65, and increased the survival rate of zebrafish. They both showed obvious inhibitory effects against xenografts of different human cancer cells including colon cancer cells (HCT116), pancreas adenocarcinoma cells (PANC-1), lung cancer cells (A549), liver cancer cells (Hep3B) and cervical carcinoma cells (Hela) in zebrafish embryos, and exhibited strong anti-bacterial effects at the concentration of 15.63 mg/mL. The two dosage forms of Forsythia Suspense have similar anti-inflammatory, antitumor and antibacterial effects, but their effects for inhibiting IL-6, P65, and Jnk mRNA expressions and HCT116 cell proliferation differ significantly at low doses in zebrafish.

In vitro evaluation and characterization of cisplatin loaded nanofibers for local chemotherapy

Cancer is a disease that affects the quality of life of the patients that are treated with Cisplatin (CDDP), which is needed for adjuvant therapy, however it leads to many secondary and adverse effects. In this study, we manufactured and characterized poly-(lactic acid) (PLA) non-woven fibers loaded with Cisplatin (CDDP) by electrospinning technique to evaluate their cytotoxicity in in vitro assays on HeLa cells (cervical carcinoma cell line). PLA–CDDP solutions with increasing concentrations of CDDP (0.5, 1 and 2% w/w) were used in a TL-01 electrospinning equipment with the same system parameters. We analyzed the chemical, thermal and morphological characteristics of PLA and PLA–CDDP fiber mats. Furthermore, hydrolytic degradation, hemolysis and toxicity in HeLa cells were evaluated. By adding the CDDP to the fibers, the degradation, glass transition and melting temperatures were modified; the 3 µm fiber diameter of pristine PLA fibers was decreased in half the size and the degradation time was extended over 5 months. However, the hemocompatibility of the material with and without CDDP was maintained, while cytotoxicity in HeLa cells increased in the three concentrations of fiber mats of PLA–CDDP compared to the intravenous drug at 24 h (p < 0.01). We concluded that the fiber mats PLA–CDDP could be used for localized therapy in the adjuvant treatment when resection panels are expose after a surgical extirpation of solid tumors.

Cassia fistula L. bark fraction modulated GSK3β/ p53 expression for mitochondrial mediated apoptosis in HeLa cells

In Ayurveda, Cassia fistula L. commonly known as Aragvadha (disease killer) is highly valuable medicinal plant. In the present study, hexane fraction (CaMH), chloroform fraction (CaMC), ethyl acetate fraction (CaME) and methanol fraction (CaMM) isolated from C. fistula bark were explored for their cytotoxic potential against cervical carcinoma (HeLa), breast carcinoma (MCF-7) and osteosarcoma (MG-63) cell lines using MTT assay. Our results unveiled that C. fistula bark fractions were selectively cytotoxic to carcinoma cells and among the tested fractions CaMC fraction had highest selectivity index (SI) with a value of 9.77 in HeLa cells. CaMC fraction actuated apoptosis in HeLa cells, as evident from chromatin condensation, DNA fragmentation, externalization of phosphatidylserine, altered mitochondrial membrane potential and ROS generation. Moreover, CaMC fraction upregulated the expression of GSK3β and p53 which modulated the Bax/ Bcl-2 ratio and initiated the release of cytochrome c from mitochondria. The expression of caspase 3/9 was also found to be upregulated which ultimately induced apoptosis. We also found that CaMC induced cell cycle arrest at G0/G1 phase and significantly inhibited the migration in HeLa cells. GC–MS analysis showed Oleic Acid, Piperine, Isopiperine and Tris(2,4-di‑tert-butylphenyl) phosphate as major constituents which could be attributed to anticancer activity of CaMC. The present study affirms the potential of CaMC fraction in modulating the aberrant signaling pathway for the induction of apoptosis in HeLa cells.

Tris-Functionalized Polyoxotungstovanadate-Mediated Antitumor Efficacy Involves Multiple Cell Death Pathways.

Polyoxometalates (POMs) are promising inorganic drug candidates for cancer chemotherapy. They are becoming attractive because of their easy accessibility and low cost. Herein, we report the synthesis and antitumor activity studies of four Lindqvist-type POMs with mixed-addenda atoms Na2[V4W2O16{(OCH2)3CR}] (R=-CH2OH, -CH3, -CH2CH3) and (Bu4N)2[V3W3{(OCH2)3CH2OOCCH2CH3}]. Compared with the current clinical applied antitumor drug 5-fluorouracil (5-FU) or Gemcitabine, analysis of MTT/CCK-8 assay, colony formation and wound healing assay revealed that the {V4W2} POMs had acceptable cytotoxicity in normal cells (293T) and significant inhibitory effects on cell proliferation and migration in three human tumor cell lines: human lung carcinoma cells (A549), human cervical carcinoma cells (HeLa), and human breast cancer cells (MCF-7). Interestingly, among the POMs analyzed, the therapeutic index (TI) of the {V4W2} POM with R= -CH2OH was relatively the most satisfactory. Thus, it was subsequently used for further studies. Flow cytometry analysis showed it prompted cellular apoptosis rate. qRT-PCR and Western blotting analysis indicated that multiple cell death pathways were activated including apoptosis, autophagy, necroptosis and pyroptosis during the POM-mediated antitumor process. In conclusion, our study shows that the polyoxotungstovanadate has great potential to be developed into a broad-spectrum antitumor chemotherapeutic drug.

Investigation of antioxidant and anticancer potential of fucoidan (in-vitro &in-silico) from brown seaweed Padina boergesenii

The present study was aimed to evaluate the antioxidant, anticancer activity and in-silico analysis of fucoidan extracted from the brown seaweed Padina boergesenii (SF-PB). The overall dry mass of the isolated SF-PB and sulphate residues was found to be 4.9 ± 0.12 % and 17.72 ± 0.25 % respectively. The Fourier Transform Infrared Spectroscopy (FTIR), X-Ray diffraction analysis (XRD), Nuclear Magnetic Resonance (NMR) Spectroscopy, High-performance liquid chromatography (HPLC), and Electrospray Ionisation Mass Spectrometry (ESI-MS) are used to further characterize the fucoidan. The antioxidant assays results showed significant antioxidant activity in a dose-dependent manner. The highest antioxidant activity was observed in the 2,2-diphenyl-1-picrylhydrazyl (DPPH)-76.75 %, Reducing power-66.55 %, 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) -(ABTS)-70.55 %, Hydrogen peroxide (H2O2)-59.9 %, Nitric oxide, and Total antioxidant scavenging (TAC) -68.92 % at 1000 μg/mL with an IC-50 value of 55.62, 59.70, 57.43, 64.40, 61.88 and 60.73 respectively. The cytotoxic effect of SF-PB on human cervical carcinoma cells (HeLa) demonstrated a significant percentage of inhibition 54 % at 40 μg/mL. Fucoidan IC 50 value against cancer cells was determined and found to be 38 μg/mL. Further, the apoptotic staining analysis depicted that fucoidan acts on HeLa cells and induces apoptosis through nuclear fragmentation, inhibition of the cell cycle at the G1 phase, and also with a high level of ROS production. Furthermore, Mesocyclops leuckarti copepod showed no toxicity when treatsed with fucoidans at a concentration of 250 μg/mL (100 % survival), demonstrating that fucoidan are not hazardous and are environmentally benign. In a docking investigation, fucoidan demonstrated the highest binding affinity against apoptotic proteins. Fucoidan may be an alternative for patients with cervical cancer. The findings showed that isolated fucoidan from P. boergesenii has substantial antioxidant and anticancer effects.

The effects of meloxicam, lornoxicam, ketoprofen, and dexketoprofen on human cervical, colorectal, and mammary carcinoma cell lines

Targeting the inflammation-related molecules with nonsteroidal anti-inflammatory drugs (NSAIDs) represents a promising approach for cancer prevention/therapy. We evaluated the in vitro anticancer effects of meloxicam, lornoxicam, ketoprofen, and dexketoprofen on the proliferation, migration, and apoptosis of human cervical, colorectal, and mammary carcinoma cells. The antiproliferative activity and cytotoxicity of tested NSAIDs on HeLa, HT-29, and MCF-7 cell lines were assessed by the MTT test. The apoptosis-inducing potential was analyzed by fluorescent staining with acridine orange/ethidium bromide and DAPI. Migration activity was assessed by a wound-healing scratch assay. The tested NSAIDs reduced the viability of the used tumor cell lines. The cytomorphological analysis revealed reduced cell density and mitotic activity and the presence of cells with morphological features of early and late apoptosis. Significant inhibition of the migration capacity was established as well. In conclusion, NSAIDs could be candidates for the development of new pharmacological strategies for the treatment and prevention of cancer.