HeLa Cell Lines Research Articles

Efficacy of graphene quantum dot-hyaluronic acid nanocomposites containing quinoline for target therapy against cancer cells

Abstract The study aims to assess the impact of graphene quantum dot-hyaluronic acid-quinoline nanocomposites (GQD-HA-Qu NCs) on MCF-7, HT-29, A2780, PANC-1, and HeLa cell lines. The GQD-HA-Qu NCs were characterized using dynamic light scattering (DLS), field emission scanning electron microscopy (FESEM), and Fourier-transform infrared (FTIR) spectroscopy. MTT assays and flow cytometry evaluated the cytotoxic and apoptotic effects of synthesized NCs. Additionally, real-time PCR was utilized to assess apoptotic gene expression. The DLS assay revealed a particle size of 224.96 nm with a polydispersity index (PDI) of 0.3. The FESEM analysis also confirmed the uniform spherical morphology of NCs. The MTT assessment demonstrated significant cytotoxicity in all cell lines, with MCF-7 and A2780 exhibiting pronounced sensitivity (P < 0.001). The flow cytometry analyses also revealed a dose-dependent increase in late apoptosis at higher concentrations of GQD-HA-Qu NCs. Notably, p53 expression was significantly upregulated compared to the untreated cells (P < 0.01), while caspases 8 and 9 showed no substantial change. This finding indicates that the p53 pathway is predominant in mediating GQD-HA-Qu NCs-induced apoptosis. The present study suggests that GQD-HA-Qu NCs are a promising treatment with selective cytotoxicity against cancer cells and robust antioxidant activity. These findings warrant further investigation for potential clinical applications.

Abstract A040: Survival mechanisms common to immunotherapy and drug tolerant persister tumor cells

Abstract Background: Persisters are a sub-population of tumor cells that survive anti-cancer therapy and are thought to be a major cause of tumor recurrence. Persisters have been identified following both drug- and immune-therapy but are generally considered to be very distinct entities. The focus of this study is to examine any similarities/differences between immunotherapy persister cells (IPCs) and drug tolerant persister cells (DTPs). Both pharmacological agents and immune cells often kill via apoptosis. Therefore, we investigated if reduced apoptotic sensitivity is a survival mechanism common to IPCs and DTPs and whether the use of BH3 mimetics to enhance apoptotic sensitivity could reduce the survival of IPCs and DTPs. Multi-therapy resistance is often observed in the clinic, therefore, we investigated if IPCs and DTPs exhibit a reciprocal multi-therapy resistant phenotype which could be conferred through a reduction in apoptotic sensitivity. Methods: IPCs were generated by co-culturing HeLa and U251 tumor cell lines with EGFR-CAR T cells for 3 weeks, replenishing CAR T cells every 3 days. Using three tumor cell line models (Hela, MC38 and B16), nine DTP cell lines were generated by chronically treating tumor cells for 7 days with an IC90 dose of drug (5-FU, oxaliplatin and etoposide), replenishing drug dose every 3 days. A CellTiter-glo or CellTox assay was used to determine the sensitivity of IPCs and DTPs to drugs and radiation. Sensitivity to T cell attack was assessed by flow cytometry (cell counting beads and zombie viability dye. Microscopy-based BH3 profiling was employed to measure the mitochondrial apoptotic sensitivity of persisters and to probe for therapeutically exploitable anti-apoptotic dependencies. Results: IPCs acquired a reduced sensitivity to multiple drug classes and radiotherapy. Likewise, DTPs also developed a reduced sensitivity to multiple drug classes and radiotherapy but also acquired a reduced sensitivity to CAR T cell killing. Depending on the cell line and therapy, some persisters developed either an increased or decreased sensitivity to mitochondrial apoptosis. Inhibition of specific anti-apoptotic dependencies using BH3 mimetics increased apoptotic sensitivity of persisters. Conclusions: Both immunotherapy and drug-tolerant persisters develop a reduced sensitivity to multiple therapeutic modalities, suggesting non-immune mechanisms are important in the survival of cancer cells after immunotherapy. These results suggest there are common mechanisms involved in producing persisters from dramatically different therapies. Our study highlights the potential of BH3 profiling to guide the selection of therapies to target persisters. Citation Format: Maria Davern, Cole J Turner, Lara Bencsics, Brenda Chan, Jasmine Yun-Tong Kung, Michael Olson, Cheyanne Walker-Williams, David Barbie, Shaili Soni, Patrick Bhola, Marcela V Maus, Anthony Letai. Survival mechanisms common to immunotherapy and drug tolerant persister tumor cells [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Functional and Genomic Precision Medicine in Cancer: Different Perspectives, Common Goals; 2025 Mar 11-13; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2025;85(5 Suppl):Abstract nr A040.

Surface‐Enhanced Raman Scattering and Cell Imaging Ability of Plasmonic Tags Based on Covalent Conjugates of Gold Nanorods With Aromatic Alkynes

ABSTRACTThe inherent benefits of Raman spectroscopy make it a suitable tool for the analysis of complex biological objects. Detection and imaging of mammalian cells is one of its development areas. Plasmonic tags endowed with alkyne reporter intended for bioimaging by surface‐enhanced Raman scattering in the silent region are presented. Gold nanorods with covalently conjugated 4‐aminotolane were obtained using dithiobis (succinimidyl propionate) and characterized by UV–Vis absorbance, transmission electron microscopy, and dynamic light scattering. Verification of surface amide bonds formation was performed with attenuated total reflectance and X‐ray photoelectron spectroscopy. Surface‐enhanced Raman scattering spectra of the tags were registered in various media and compared with a feedback from the similar system obtained through non‐specific adsorption of 4‐aminotolane on gold nanorods. Cytotoxicity of Raman reporter itself and its conjugates with gold nanorods were verified on HeLa cell line. Spectral mapping and scanning electron microscopy revealed the localized intracellular accumulation of the tags. The challenge associated with reduced imaging area due to formation of the tags aggregates inside the cells is pointed out.

Grandifolins A-D: three 6/6/5 abietane diterpenes and an abietane diterpene from Salvia grandifolia.

The genus Salvia is a well-established source of biologically active compounds with beneficial health effects. In this study, we aimed to isolate and structurally characterize novel compounds from Salvia grandifolia and evaluate their cytotoxic activity against human promyelocytic leukemia (HL-60) and cervical cancer (HeLa) cell lines. Three new rearranged abietane-type diterpenes, grandifolins A-C (1-3) and a new abietane diterpene, grandifolin D (4), were isolated from the roots of Salvia grandifolia, along with 12 known compounds (5-16). Their structures were determined by combining extensive 1H and 13C spectroscopy, X-ray crystallography, and electronic circular dichroism spectra (ECD). The skeletons of 1-3 had a rearranged structure with a 6/6/5 ring from an abietane-type diterpene. A plausible biosynthetic pathway for the rearranged skeleton was also proposed. Many of the isolated compounds showed weak cytotoxicity against HeLa or HL-60 cell lines.

Characterization of the Cytotoxic Effect of Naphthalenacetamides Hydrochlorides on Cervical Cancer-Derived Cells.

Cervical cancer is a global health problem due to its high incidence and prevalence in women, mainly in third-world countries. For the treatment of this disease, there are different therapeutic options, but these are not always effective, which gives rise to the search for new compounds using cheminformatics tools. The objective of this study was to design, synthesize, and biologically evaluate N-(2- morpholinoethyl)-2-(naphthalen-2-yloxy)acetamide hydrochloride (1) and 2-(naphthalen-2-yloxy)- N-(2-(piperidin-1-yl)ethyl)acetamide hydrochloride (2) on the HeLa cell line in vitro. The referenced cell line from the American Type Culture Collection (ATCC®CCL-2™) was used, and the effect on cell viability was determined by MTT metabolic reduction-based assay at 24, 48, and 72 h. Therapies directed at the σ1 receptor may be a treatment alternative since this receptor modulates the processes of cell proliferation and angiogenesis, producing cytoprotective or cytotoxic actions depending on the ligand with which it is coupled. The analysis showed that compounds 1 and 2 presented activity on HeLa cancer cells and viability at micromolar concentrations (1.923 μmol/mL and 0.374 μmol/mL, respectively). Moreover, the effect was maintained for 72 h. Naphthaleneacetamide derivatives exhibited an inhibitory effect on the HeLa cell line, and the OSIRIS program predicted less toxicity than cisplatin.

A novel de novo GFAP variant causes a juvenile-onset Alexander disease with bilateral vocal cord paralysis.

Alexander disease (AxD), an autosomal dominant leukodystrophy, is caused by mutations in the GFAP, the gene encoding glial fibrillary acidic protein (GFAP). The disease, classified by age of onset into infantile, juvenile, and adult forms, is characterized by white matter degeneration and astrocytic inclusions called Rosenthal fibers. A patient underwent clinical, radiological, and molecular analyses to confirm a suspected diagnosis of AxD. The functional effect of the variant identified was tested using computational tools and in HeLa and astrocytoma cell lines. We report a case of juvenile AxD that clinically developed acute respiratory distress due to bilateral vocal cord paralysis. Brain and spinal cord MRI revealed the typical findings of the disease, including bulbospinal atrophy and T2-weighted hyperintensities in the frontal periventricular white matter. Molecular genetic testing identified a novel de novo c.713 T > G (p.I238S) variant of GFAP. In silico analyses revealed that the variant at evolutionarily conserved residue likely affects protein function. In vitro assays confirmed its pathogenic effect, showing that p.I238S protein expression significantly associates with aggregate formation in cellular models. Extending the clinical and molecular characterization of new cases of AxD is an important achievement to better characterize the disease.

Inhibitory effect of Curcumin on a cervical cancer cell line via the RAS/RAF signaling pathway.

Cervical cancer has a very important place in female infertility and ranks fourth among cancers affecting women. Curcumin (CUR) is closely associated with the expression and activity of various regulatory proteins. It is also known that curcumin has preventive and therapeutic effects on various types of cancer. In this study, the anticancer activities of curcumin were demonstrated in the human cervical cancer cell line (HeLa). qRT-PCR and western blot analyses were used to evaluate mRNA and protein expression of curcumin in HeLa and immortalized human skin keratinocyte cell lines (HaCaT) (proliferation and apoptosis regulatory markers of the RAS/RAF signaling pathway). MTT analysis was performed, showing HeLa and HaCaT cell proliferation depending on the dose and duration of curcumin and doxorubicin. A wound scratch healing assay was applied to examine cell migration and invasion of HeLa after curcumin application. To determine the role of curcumin and doxorubicin in the apoptosis of HeLa cells, the mRNA levels of caspase-3 were examined by qRT-PCR. The results were analyzed with a one-way ANOVA SPSS 20.0 program. CUR (IC50: 242.8 μM) and DOX (IC50: 92.1 μM) were determined to have the ability to inhibit the proliferation of HeLa cells and induce apoptosis over a 72-hour period and dose-dependently. Moreover, the results revealed that the mRNA and protein expression levels of RAF and RAS in HeLa cells were downregulated by CUR and DOX. The findings show that an alternative treatment method for cervical cancer can be developed with the application of CUR and DOX. Alternative methods for cervical cancer treatment may be developed using different methods in future studies.

The HGF/Met Receptor Mediates Cytotoxic Effect of Bacterial Cyclodipeptides in Human Cervical Cancer Cells.

Human cervix adenocarcinoma (CC) caused by papillomavirus is the third most common cancer among female malignant tumors. Bioactive compounds such as cyclodipeptides (CDPs) possess cytotoxic effects in human cervical cancer HeLa cells mainly by blocking the PI3K/Akt/mTOR pathway and subsequently inducing gene expression by countless transcription regulators. However, the upstream elements of signaling pathways have not been well studied. To elucidate the cytotoxic and antiproliferative responses of the HeLa cell line to CDPs by a transcriptomic analysis previously carried out, we identified by immunochemical analyses, differential expression of genes related to the hepatocyte growth factor/mesenchymal-epithelial transition factor (HGF/MET) receptors. Furthermore, molecular docking was carried out to evaluate the interactions of CDPs with the EGF and MET substrate binding sites. Immunochemical and molecular docking analyses suggest that the HGF/MET receptor participation in CDPs cytotoxic effect was independent of the protein expression levels. However, protein modulation of downstream Met-targets occurred due to the inhibition of phosphorylation of the HGF/MET receptor. Results suggest that the antiproliferative and cytotoxicity of CDPs in HeLa cells involve the HGF/MET receptor upstream of PI3K/Akt/mTOR pathway; assays with the human breast cancer MCF-7 and MDA-MB-231cell lines supported the finding. Data provide new insights into the molecular mechanisms involved in CDPs cytotoxicity and antiproliferative effects, suggesting that the signal transduction mechanism may be related to the inhibition of the phosphorylation of the EGF/MET receptor at the level of substrate binding site by an inhibition mechanism similar to that of Gefitinib and Foretinib anti-neoplastic drugs.

Using Cyclic Ketones to Synthesize New 3,5,6,7,8,9-Hexahydro-pyrazolo[1,5-a]quinoline Derivatives with Antiproliferative Activities: Morphological Studies.

Quinoline derivatives, often incorporating other heterocyclic structures, have shown a wide range of therapeutic potential, especially in the treatment of cancer. These compounds have demonstrated significant anticancer activity against various cell lines, including HeLa (human cervical cancer) and MDA-MB-435 (melanoma), exhibiting strong inhibitory effects. In this study, arylhydrazonopyrazole derivatives (3a-c) were employed in a series of multicomponent reactions to synthesize 3,5,6,7,8,9-hexahydropyrazolo[1,5-a]quinoline and pyran derivatives. Pyrazolo[1,5-a]quinoline derivatives, due to their structural properties, are considered valuable scaffolds for the development of novel drugs targeting similar biological pathways, with the potential for improved therapeutic efficacy. This study aimed to demonstrate the use of simple arylhydrazonopyrazole derivatives in multicomponent reactions with cyclic ketones and aromatic aldehydes. The resulting compounds were then assessed for their cytotoxic and antiproliferative activities. Following these reactions, further heterocyclization processes were conducted, incorporating the quinoline moiety into the final structures. These findings underscore the potential of pyrazolo [1,5-a]quinoline derivatives as promising candidates for drug discovery, offering new avenues for targeting diseases with related molecular mechanisms. The key starting compound in this study was 3,5-dimethyl-4-(2-phenylhydrazono)-4H-pyrazole, which has been utilized in numerous heterocyclization reactions. These reactions, involving various reagents, such as cyclic ketones and diketones in the presence of aromatic aldehydes, led to the formation of fused tetracyclic compounds. Arylhydrazonopyrazole derivatives (5a-c) were employed in multicomponent reactions to synthesize 3,5,6,7,8,9-hexahydropyrazolo[1,5-a]quinoline and pyran derivatives. The reactions were carried out using both conventional catalysts and ionic liquid-immobilized catalysts. Notably, the use of ionic liquid-immobilized catalysts resulted in higher yields of the desired compounds. In this study, new compounds were synthesized, characterized, and evaluated for their cytotoxicity against six cancer cell lines: A549, HT-29, MKN-45, U87MG, SMMC-7721, and H460. Additionally, the cytotoxic effects of the synthesized compounds were assessed against hepatocellular carcinoma (HepG2) and cervical carcinoma (HeLa) cell lines. Morphological studies of selected compounds were also conducted to further understand their effects on the cancer cells. Moreover, the cytotoxicity of the selected compounds was tested against seventeen different cancer cell lines, categorized by disease type. Morphological analyses of these selected compounds were also performed to gain deeper insights into their potential as anticancer agents. The inhibition assays of the tested compounds demonstrated significant activity against c-Met enzymatic activity, with IC50 values ranging from 0.25 to 10.30 nM. Additionally, potent inhibition was observed in the prostate PC-3 cell line, with IC50 values ranging from 0.19 to 8.62 μM. These promising results highlight the potential of these compounds and encourage further research to explore their therapeutic applications in the future.

LncRNA HIF1A-AS2 acts as an oncogene to regulate malignant phenotypes in cervical cancer

BackgroundLong noncoding RNAs (lncRNAs) HIF1A-AS2 is upregulated in multiple human cancers and are associated with various aspects of tumor progression. However, the molecular mechanisms of HIF1A-AS2 in cervical cancer (CC) remain largely unknown. In this study, we aim to investigate the expression pattern and signaling pathways of HIF1A-AS2 in CC.MethodsThe study included a group of 20 CC patients, from whom tumor tissue specimens were collected. Additionally, three distinct CC cell lines (HeLa, SiHa, CaSki) were utilized. Quantitative real-time PCR (qRT-PCR) was used to assess the transcript levels of HIF1A-AS2 in these samples. Functional studies were performed by CCK-8, Transwell and Apoptosis assays. Databases including JASPAR, miRDB and Targetscan were used for the transcription factor or target miRNA prediction, subsequent dual luciferase activity assay, chromatin immunoprecipitation (ChIP) and Ago2 immunoprecipitation (RIP) were also adopted for validation.ResultsThe study demonstrated that HIF1A-AS2 expression was elevated in clinical cervical cancer specimens and cultured cell lines in comparison to normal controls. Knockdown of HIF1A-AS2 notably inhibited the proliferation and invasion of cervical cancer cells, while inducing apoptosis. In contrast, HIF1A-AS2 overexpression promoted cellular proliferation and invasion and suppressed apoptosis. It was also identified that c-Jun functions as a transcription factor, activating HIF1A-AS2 expression. Additionally, HIF1A-AS2 was found to serve as a molecular sponge for miR-34b-5p, negatively regulating its expression. Furthermore, HIF1A-AS2 controlled the expression of radixin (RDX) by sponging the miR-34b-5p pathway.ConclusionOur findings indicate that c-Jun-activated HIF1A-AS2 acts as an oncogenic factor in CC by sponging miR-34b-5p to target radixin. These findings suggest that HIF1A-AS2 might be a viable and promising therapeutic target for cervical cancer treatment.

Asperacunoid A, a new cytotoxic germacrane-type sesquiterpenoid isolated from Aspergillus aculeatinus 2022EX723

Sesquiterpenoids have obtained widespread concern because of their intriguing molecular architecture and considerable bioactivities. In the present study of chemical diversity of Aspergillus aculeatinus 2022EX723, an endophytic species derived from the marine green algae Enteromorpha prolifera, three sesquiterpenoids including a new germacrane-type sesquiterpenoid asperacunoid A (1) and two sesquiterpenoids (2 and 3) were isolated and identified. With the assistance of spectroscopic analyses (HRESIMS, NMR, and X-ray analyses), the structures as well as the absolute configuration of 1 were successfully elucidated. Cytotoxic assays against A549, HeLa, HXO-Rb44, and MKN-45 cell lines revealed that asperacunoid A (1) was active against HXO-Rb44 and MKN-45 cells, with respective IC50s of 10.9 and 23.8 μM (compared with the positive control doxorubicin with IC50s of 11.2 μM for HXO-Rb44 and 13.3 μM for MKN-45 cells). This study indicated that the new sesquiterpenoid 1 may be served as a lead compound with high pharmacological potentiality.

Bio-fabricated silver nanoparticles: therapeutic evaluation as a potential nanodrug against cervical and liver cancer cells

Nanobiotechnology has grown rapidly and is now widely used in the diagnosis and treatment of modern diseases. Biosynthesized silver nanoparticles (AgNPs) are eco-friendly, cost-effective, biocompatible route, and have biomedical properties at minimal concentrations. In the present study extract of cyanobacterium (Anabaena variabilis) was utilized to synthesize facile, reliable AgNPs, further biosynthesized AgNPs were characterized by physicochemical techniques. The atomic force microscope study confirmed the shape of AgNPs while the scanning electron microscopy study revealed 17 to 35 nm in the size range. The zeta potential value of -19.5 mV demonstrated the repulsion effect between the particles, which prevents their aggregations while the heating stability of AgNPs was confirmed by Thermogravimetry differential thermal analysis. Another important characteristic, such as elemental constituent of AgNPs was determined by inductively coupled plasma mass spectrometry and was observed 94.24, 95.19, 97.06, and 99.34% of silver present in their respective concentrations of AgNPs. In vitro cytotoxicity of AgNPs was screened on HeLa, SiHa, (Cervical carcinoma), and HepG2 (Human hepatocellular carcinoma), cell lines. To evaluate the biocompatibility of AgNPs immortalized human embryonic kidney (HEK-293) cell line was used. The IC50 values of AgNPs are were observed as 23.76 ± 2.4 µg/mL, 11.21 ± 1.7 µg/mL, and 22.27 ± 1.8 µg/mL against HeLa, SiHa and HepG2 cell lines respectively. AgNPs demonstrated the biocompatible nature against HEK-293 cells, Normal cell line (HEK-293) cytotoxicity results showed exhibited ≥ 95% cell viability at all the concentrations. During the DAPI (4’,6-diamidino-2-phenylindole) staining study IC50 dose of AgNPs on cancer cell lines (HeLa, SiHa, and HepG2) showed nuclear morphological alterations which indicate the DNA damage and apoptosis in cancer cells. AgNPs treated cancer cells increased the cells number in the S phase while decreased the number of cells in the G0-G1 and G2/M phases of the cell cycle in all three cancer cells compared to the control.Graphical

5-Chlorosalicylaldehyde Based New Chromo-Fluorogenic Schiff base Probes for Cu(II), Mn(II), CN-, S2- and CO32- ions - Appositeness in Bioimaging and Molecular Logic Gate.

Two new multifunctional fluorescent Schiff base chemosensors 2,2'-(4-chlorophenyl)methylene)bis(azanylylidene))bis(methanylylidene))bis(4-chlorophenol) (CSCB) and 2,2'-(4-methoxyphenyl)methylene)bis(azanylylidene))bis(methanylylidene))bis(4-chlorophenol) (CSMB) were synthesized by following solvent free one pot synthesis approach for the colorimetric detection of Cu(II), Mn(II), CO32-, S2- and CN- ions. The Schiff base chemosensors showed a colorimetric response towards the ions, where there was appearance of yellow color over colorless Schiff base solution. The detection process was accomplished by absorption and emission methods in aqueous methanol medium. The binding modes of the cations and anions towards both the Schiff bases were explored by IR, 1H NMR and Mass titrations. The detection limit of the Schiff bases CSCB and CSMB towards the different detected ions was found to be in the order of 10-6M for Cu(II), Mn(II), CN-, CO32- ions and 10-5M for S2- ion. DFT studies of the Schiff bases CSCB and CSMB were performed with respect to Mulliken charge analysis, HOMO-LUMO energy gap, Molecular Electrostatic Potential and Total density for the substantial support of structure and interaction with various ions. Electronic spectral transitions of the Schiff bases are endorsed by TDDFT calculations. The sensing property was applied in bioimaging study on HeLa cell lines and logic gate construction.

Limonoid-rich fraction from Azadirachta indica A. Juss. (neem) stem bark triggers ROS-independent ER stress and induces apoptosis in 2D cultured cervical cancer cells and 3D cervical tumor spheroids

BackgroundThe existing anticancer drugs in clinical practice show poor efficacy in cervical cancer patients and are associated with multiple side effects. Our previous study demonstrated the strong antineoplastic activity of crude extract prepared from the stem bark of Azadirachta indica (Neem) against cervical cancer. However, the active phytoconstituents of neem stem bark extract and its underlying anticancer mechanism are yet to be investigated. Thus, the present study aimed to identify the active fraction from crude neem stem bark extract to further dissect its anticancer mechanism and determine the active components.MethodsDichloromethane (DCM) extract from neem stem bark was prepared and fractionated using thin-layer chromatography. The fractions obtained were screened against HeLa and ME-180 cervical cancer cell lines to identify the most active fraction, which was then selected for further studies. Clonogenic assay, cell cycle analysis, apoptosis assay, and reactive oxygen species (ROS) assay were performed to determine the cytotoxicity of the active fraction. Gene expression was analyzed using real-time PCR and western blot to determine the mechanism. Additionally, the HeLa cells-derived 3D spheroid model was used to determine the antitumor efficacy of the active fraction. Electrospray ionization-mass spectrometry, Fourier-transform infrared spectroscopy, and proton nuclear magnetic resonance were used to identify the phytoconstituents of the fraction.ResultsInitial screening revealed fraction 2 (F2) as the most active fraction. Additionally, F2 showed the least cytotoxic effect on normal human fibroblast cells. Mechanistically, F2 induced cell cycle arrest and apoptosis in cervical cancer cells. F2 increased ROS levels, induced ER stress, and activated cell survival pathway. Treatment with N-acetyl cysteine revealed that F2 induced ROS-independent ER stress and apoptosis. 3D spheroid viability and growth delay experiments demonstrated the strong antitumor potential of F2. Finally, six compounds, including one flavonoid (nicotiflorin) and five limonoids, were identified in the F2 fraction.ConclusionThis is the first study to identify the active fraction and its phytoconstituents from neem stem bark and demonstrate the anticancer mechanism against cervical cancer. Our study highlights the importance of investigating neem stem bark-derived limonoids and nicotiflorin as a potential source to develop new anticancer therapeutic agents.Graphical

Capsicum chinense Jacq. fruit plays an immunomodulatory role in cytokine attenuation and DNA damage protection.

Capsicum chinense (C. chinense) Jacq., recognized for its bioactive compounds, has attracted interest due to its possible immunomodulatory and DNA-damage-protective effects. This study aimed to assess the immunomodulatory, anti-inflammatory, and DNA protection abilities of organic extracts (methanol, ethyl acetate, and petroleum ether) from C. chinense. The immunomodulatory effects were evaluated in Long Evans rats induced with SRBC. At the same time, the anti-inflammatory potential was investigated in LPS-stimulated RAW264.7 macrophages by quantifying pro-inflammatory mediators such as COX-2, iNOS, TNF-α, IL-β, and IL-6. Protective activity against DNA damage was assessed using a method that induces damage with a Fenton reagent. Cytotoxicity was tested on the Hela cell line to gauge the cellular effects of the extracts. The results demonstrated that higher doses (200 mg) of C. chinense methanol extract inhibited immune responses, whereas lower doses helped restore them. The extracts significantly decreased pro-inflammatory mediators and cytokines in LPS-activated macrophages. Both petroleum ether and methanol extracts showed higher cytotoxicity against Hela cells compared to the ethyl acetate extract. The protein levels recorded in the serological parameters were 5.74 ± 0.19, 5.36 ± 0.11, 5.74 ± 0.26, 6.02 ± 0.16, 6.18 ± 0.18, and 6.22 ± 0.20 gm/dL for NC, DC, ML, MLMExCC100, MLMExCC200, and MExCC100, respectively. These findings suggest that C. chinense extracts possess strong immunomodulatory effects and potential protection against DNA damage, supporting their therapeutic application in regulating the innate immunity system.

CHARACTERIZATION, EVALUATION OF CYTOTOXIC ACTIVITY ASSAY AND OPTIMIZATION OF NANOEMULSION DELIVERY SYSTEM FORMULAS OF TOMATO LYCOPENE (SOLANUM LYCOPERSICUM. L)

Objective: This study aimed to evaluate the cytotoxic effects of tomato lycopene, quantify its bioactive fraction, and formulate tomato lycopene into nanoemulsions. Methods: Lycopene was fractionated using chloroform and purified using methanol. Tomato lycopene was identified using liquid chromatography-mass spectrometry and ultraviolet-visible spectrophotometry, with the content determined using thin-layer chromatography-densitometry. Cytotoxicity was assessed in T47d, DU145, and HeLa cells using the MTT assay. The composition of the oil phase (Virgin Coconut Oil), surfactant (Tween 80), cosurfactant (PEG 400), and the aqueous phase of the nanoemulsion base was determined using a pseudo ternary phase diagram. Tomato lycopene was added to the oil phase and mixed with other components by spontaneous titration. The nanoemulsions were characterized by determining droplet size, zeta potential, Poly Dispersity Index (PDI), transmittance, pH, density, and morphology using transmission electron microscopy. Results: Needle-shaped crystals were obtained, with a retention factor of 17.32 min, m/z 535.4316 (calculated for C40H56), and maximum wavelengths of 457, 484, and 517 nm. The bioactive fraction (chloroform) comprised 866.68 mcg/ml lycopene. Increasing lycopene concentration was inversely proportional to T47d, DU145, and HeLa cell viability after 96 h of incubation. Six of the 54 base formulations produced transparent solutions (droplet size: 14.10–500.50 nm). Incorporating 0.1% tomato lycopene into the base generated physically stable nanoemulsions with spherical droplets exhibiting the following features: particle size, 13.37–82.52 nm; zeta potential, (-12.4)–(-5.66) mV; PDI, 0.0813–0.4247; transmittance, 96.18–99.14%; pH, 5.49–6.40; relative density, 1.049–1.067. Conclusion: Tomato lycopene showed weak citotoxic on T47d, DU145, and HeLa cell line. Six nanoemulsions with good physicochemical properties were obtained as transparent yellow solutions. The optimized lycopene nanoemulsion formulation (TLN6) was confirmed at the compisition of 0,1% lycopene, 7% VCO, 50,4% Tween 80 and 12,6% PEG 400.

Effect of 6,6-Substitution on 2,2'-Bipyridine and Probing Biomolecular Interaction Through Experimental and Computational Studies Towards Anticancer Potency.

The current study focuses on the cytotoxicity assessment of 6,6'-substituted 2,2'-bipyridine derivatives containing functional groups, namely methyl (-CH3, L1), acid (-COOH, L2), ester (-COOCH3, L3), semicarbozone (-CONHNH2, L4). The ligands exhibited high solubility in dimethyl sulfoxide (DMSO) and moderately soluble in dimethyl formamide (DMF), acetonitrile (ACN) and low solubility in water. The ligands display a significant π→π* transition in the region of 270nm to 305nm. The emission spectra of the ligands reveal a prominent band in the 300nm to 450nm range, with a Stokes shift of 120cm- 1. UV-VIS spectra analysis, the ligands (L1-L4, 1mM) demonstrated stability in various environmental like water, glutathione (GSH, 1mM), and MTT condition (10% DMSO). The interaction of ligands with DNA was assessed through calf thymus deoxyribonucleic acid (ctDNA) binding assay, viscosity studies, and ethidium bromide (EtBr) displacement assay, with L1 and L4 exhibiting the highest interaction. The MTT assay was conducted for 24h and 48h using ligands (L1-L4) and doxorubicin in MCF-7, HeLa, and L929 cell lines. Ligand L4 showed high potency IC50 values after 48h in MCF-7 (1.28 µM) and HeLa (1.81 µM), but its potency is lower than doxorubicin. Overall, for the first time these results suggest that L4 has high anticancer activity at lower concentrations against breast and cervical cancers.

Therapeutic implications of endoplasmic reticulum stress gene CCL3 in cervical squamous cell carcinoma

This study investigated ERS-related gene expressions in CESC, identifying two molecular subtypes, P1 and P2, and constructing a precise prognostic model based on these subtypes. TCGA's whole-genome expression profiles were used to recognize these subtypes through the ConsensusClusterPlus method, further refining prognostic models with univariate and Lasso Cox regression analyses validated by the GSE39001 dataset. The study analyzed the expression distribution of ERS marker genes within T cell subgroups using scRNA-seq data (GSE168652), highlighting T cell diversity. The critical role of the CCL3 gene in prognostic models was examined explicitly in CD8 + T cells from healthy individuals and CESC patients. Elevated CCL3 levels were observed in patients' CD8 + T cells compared to healthy controls. Functional experiments involving CCL3 knockdown and overexpression in HeLa and SiHa CESC cell lines were conducted to investigate its impact on cell proliferation, migration, and invasion. These findings were subsequently validated in a nude mouse model. The results demonstrated that suppressing CCL3 inhibited cell proliferation, migration, and invasion significantly, while its overexpression promoted these processes. In the mouse model, CCL3 silencing reduced tumor growth and decreased Ki-67 labeling within the tumor tissues, indicating the therapeutic potential of targeting CCL3 in CESC treatment, possibly through CD8 + T cell regulation. This study contributes new prognostic assessment tools and personalized treatment options for CESC patients, paving the way for more targeted therapies in CESC by discovering the CCL3 gene, presenting significant clinical implications.

Synergistic Antiproliferative Effect of Linagliptin-Metformin Combination on the Growth of Hela Cancer Cell Line

This in-vitro study explores the cytotoxic properties of the linagliptin-metformin combination on cervical cancer cells and examines the synergistic interaction between the two drugs. An MTT assay was used to explore the anti-cancer effects of the linagliptin-metformin mixture on a cervical cancer cell line (HeLa cell line) across 24 and 72-hour incubation periods. The concentrations of metformin, linagliptin, and their combination ranged from 0.1 to 1000 µg/ml. while the concentrations in the mixture were kept at fifty percentage of the individually used drug. The study included an estimated combination index value (CI) and the dosage reduction index (DRI) to ascertain the possibility of a synergistic effect between combined drugs and mixture safety. study finding exhibited that all studied drugs- metformin, linagliptin, and their combined mixture- inhibited the growth of cervical cancer cells with a superior efficacy of the mixture over individual drugs. Inhibition patterns of the drugs were directly proportional to the drug's concentration and the incubation time. The combination index finding revealed that the mixture's cytotoxic effect of metformin and linagliptin was synergistic. The dose reduction index value revealed that lower drug concentrations were required in the combination mixture than when used individually indicating a greater cytotoxic potential of the mixture. The study findings of MTT, CI, and DRI indicate that the mixture is an effective, safer, and promising anticancer therapy for cervical cancer. Conclusion: This study explores the cytotoxic potential of metformin and linagliptin individually and in combination. The greater cytotoxic potential of the drugs in combination highlights their lower effective concentrations, paving the way for further research on using these drugs for effective cancer treatment.

Eco-friendly synthesis of ZnO, CuO, and ZnO/CuO nanoparticles using extract of spent Pleurotus ostreatus substrate, and their antioxidant and anticancer activities

Biosynthesis techniques for nanomaterials have advanced significantly, promoting eco-friendly synthesis chemistry as a sustainable alternative to conventional methods. This study presents a novel and environmentally friendly approach for synthesizing nanoparticulate ZnO, CuO, and ZnO/CuO nanocomposites using aqueous extracts of Pleurotus ostreatus spent substrate, is reported. The structural, optical, and morphological properties of the synthesized NPs were analysed. A hexagonal phase of ZnO NPs and a monoclinic phase of CuO NPs were obtained according to the X-ray diffraction analysis. A reduction in the peak intensity of these metal oxides was observed in the ZnO/CuO NPs due to reduced crystallinity. The absorption spectra, obtained from the UV–vis analysis, showed peaks at 354, 365, and 525 nm for the ZnO, CuO, and ZnO/CuO NPs, respectively. An anticancer assay of the NPs was conducted using human embryonic kidney (HEK 293) and cervical carcinoma (HeLa) cell lines, while a 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay was used for the antioxidant evaluation. The ZnO, CuO, and ZnO/CuO NPs showed higher antioxidant potency with IC50 of 2.15, 2.16, and 3.18 µg/mL, respectively, than the ascorbic acid (4.25 µg/mL). This indicates that the nanoparticles were more effective in capturing DPPH free radicals. Anticancer assays showed strong cytotoxic effects for all nanoparticles, with ZnO NPs exhibiting the highest activity (IC50: 1.94 μM for HEK 293 cells, 3.23 μM for HeLa cells), surpassing CuO and ZnO/CuO NPs. Cell viability for both HEK 293 and HeLa cells decreased as nanoparticle concentration increased, confirming dose-dependent cytotoxicity. The green synthesized metal oxides and their composite have the potential for biomedical applications.