IC50 Dose Research Articles

Morphological analysis and cytotoxicity of acrylamide on SPC212 human mesothelioma cells: Do low doses induce proliferation, while high doses cause toxicity?

AbstractAcrylamide is broadly utilized in numerous areas with different purposes including being an additive, flocculating, sealing, dry strength improver and polymerizing agent, and so forth. Furthermore, it forms in certain food products at high temperatures. It poses serious hazard since its readily water‐soluble and very reactive nature. Besides in vivo studies, several in vitro studies with various cell lines are carried out to evaluate its toxicity. However, of these cell line studies, there are no mesothelium or mesothelioma cell lines. To fill this lacuna, we aimed at examining various dose range of acrylamide on SPC212 human mesothelioma cell line. First, we executed MTT and neutral red cytotoxicity tests and ascertained IC50 dose. Next, we performed inverted, light (haematoxylin–eosin and May Grünwald), fluorescent (DAPI) and confocal microscope (AO/EB) analyses as well as immunohistochemistry for Bax, Bcl‐2 and PCNA proteins. As a result, we found IC50 of acrylamide at 2.65 mM. Starting from 3.13 mM of acrylamide dose, a deep decrease in cell proliferation was observed. Particularly in MTT assay, a proliferative action of acrylamide was detected at 0.39 and 0.78 mM, supported with inverted microscope images. In light microscope analysis, several cellular degenerations, including condensed and kidney‐shaped nucleus were evident. In AO/EB staining, cells with apoptotic characteristics augmented dose‐dependently, being upheld by a parallel uptick in Bax and a dimunition in Bcl‐2 staining. Besides, PCNA decreased at IC50 dose of acrylamide. This is the acrylamide‐associated first study conducted on SPC212 mesothelioma cells encompassing advanced morphological analysis. We believe this study to be an incentive for future studies.

Susceptibility of Leishmania amazonensis Axenic Amastigotes to the Calpain Inhibitor MDL28170

Leishmaniasis encompasses a group of neglected diseases caused by flagellated protozoa belonging to the Leishmania genus, associated with high morbidity and mortality. The search for compounds with anti-Leishmania activity that exhibit lower toxicity and can overcome the emergence of resistant strains remains a significant goal. In this context, the calpain inhibitor MDL28170 has previously demonstrated deleterious effects against promastigote forms of Leishmania amazonensis, which led us to investigate its role on axenic amastigote forms. The calpain inhibitor MDL28170 was able to decrease the viability of amastigotes in a typically dose-dependent manner. The treatment with the IC50 dose (13.5 μM) for 72 h led to significant amastigote lysis and increased cell-to-cell aggregation. Ultrastructural analysis revealed several cellular alterations, including disruption of the trans-Golgi network and the formation of autophagosomes when treated with MDL28170 at ½ × IC50 dose. Additionally, mitochondrial swelling and the formation of concentric membranous structures inside the mitochondrion were observed after incubation with the IC50 dose. These results reinforce the potential application of the calpain inhibitor MDL28170 against L. amazonensis, highlighting its effectiveness and possible mechanism of action against the parasite.

Napabucasin Inhibits Proliferation and Migration of Glioblastoma Cells (U87) by Regulating JAK2/STAT3 Signaling Pathway.

Background and Objectives: Napabucasin (NP) was discovered as a natural compound that suppresses cancer stemness by inhibiting the signal transducer and activator of the transcription 3 (STAT3) signaling pathway. In this study, the anti-proliferative and apoptotic effects of NP and the chemotherapy agent doxorubicin (DX), a natural compound, on glioblastoma cells (U87) were investigated. Materials and Methods: In this study, the effects of NP and DX on cell viability on the glioblastoma U87 cell line were determined by MTT test. Expressions of Jak2/Stat3 genes were examined by qRT-PCR. Apoptosis was evaluated by Hoescht 33258 staining. Moreover, NP, its antagonistic-synergistic effects and IC50 doses of the combined treatment of DX were determined. Results: Napabucacin and doxorubicin were found to inhibit glioblastoma U87 cell proliferation. It was determined that NP applied in the range of 0.3-1 µM and its combination with DX killed almost all of the glioblastoma cells in 48 h of application. Additionally, it was observed that Jak2/Stat3 expressions downregulated. Conclusions: These results show that NP suppresses the proliferation of glioblastoma cells. It was shown that the combination of NP and DX can prevent invasion of the U87 cell line due to its Jak2/Stat3 inhibitory effect. Since it can suppress Jak2/Stat3, an important cancer cell proliferation pathway in glioblastoma, the combination of NP and DX can be used as an alternative treatment agent. But no synergistic effect of NP and DX on the U87 cells of the glioblastoma cell line was observed.

Chemical analysis and bioactivity evaluation of Citrus limon leaves volatile oil from Palestine: investigating phytochemical, anti-inflammatory, antimicrobial, and cytotoxic properties

IntroductionScientists consider repurposing discarded waste into valuable resources for the pharmaceutical, cosmetic, and food flavoring manufacturing industries a challenge to generate new revenue streams, lower production costs, and reduce waste. Therefore, the current study aimed to identify Citrus limon leaf volatile oil (VO) constituents from Palestine and assess its in vitro antimicrobial, anti-inflammatory, and cytotoxic properties. MethodsThe components of the VO were analyzed using Gas Chromatography/Mass Spectrometry (GC/MS). The antimicrobial activity of VO was assessed using a microdilution technique. The cyclooxygenases (COX-1 and COX-2) assay was utilized to assess the anti-inflammatory effect. An aqueous one solution cell proliferation (MTS) assay was employed to determine the cytotoxic effect of the VO. ResultsThirty-six molecules were identified in the oil, and geranial, neral, and limonene are the most abundant molecules, comprising 31.06%, 23.98%, and 14.32%, respectively. The antimicrobial results showed that the VO has notable inhibition against Proteus vulgaris, Methicillin-resistant Staphylococcus aureus, and Candida albicans. The COX IC50 calculations revealed that it has high potency against COX-2 IC50=9.32±0.88µg/ml. The most potent cytotoxic effect of C. limon VO was noticed against MCF-7, with an IC50 dose of 162.90±1.85 mg/ml. ConclusionThe GC-MS analyses revealed that geranial, neral, and limonene are the predominant compounds in the VO of C. limon leaves from Palestine. The biological test results demonstrated that the VO has potential inhibitory actions against some cancer cells, bacterial and fungal species, and COX-2 enzyme, suggesting it may be a viable choice for treating or preventing microbial infections, cancer, and inflammatory illnesses.

Development of novel dual-target drugs against visceral leishmaniasis and combinational study with miltefosine

The dual-target inhibitors (ZINC000008876351 and ZINC000253403245) were identified by utilizing an advanced computational drug discovery method by targeting two critical enzymes such as FeSODA (Iron superoxide dismutase) and TryR (Trypanothione reductase) within the antioxidant defense system of Leishmania donovani (Ld). In vitro enzyme inhibition kinetics reveals that both the compound's ability to inhibit the function of enzyme LdFeSODA and LdTryR with inhibition constant (Ki) value in the low μM range. Flow cytometry analysis, specifically at IC50 and 2X IC50 doses of both the compounds, the intracellular ROS was significantly increased as compared to the untreated control. The compounds ZINC000253403245 and ZINC000008876351 exhibited strong anti-leishmanial activity in a dose-dependent manner against both the promastigote and amastigote stages of the parasite. The data indicate that these molecules hold promise as potential anti-leishmanial agents for developing new treatments against visceral leishmaniasis, specifically targeting the LdFeSODA and LdTryR enzymes. Additionally, the in vitro MTT assay shows that combining these compounds with miltefosine produces a synergistic effect compared to miltefosine alone. This suggests that the compounds can boost miltefosine's effectiveness by synergistically inhibiting the growth of L. donovani promastigotes. Given the emergence of miltefosine resistance in some Leishmania strains, these findings are particularly significant.

Copper nanoparticles biosynthesis by Priestia megaterium and its application as antibacterial and antitumor agents

The growth of material science and technology places high importance on creating better processes for synthesizing copper nanoparticles. Thus, an easy, ecological, and benign process for producing copper nanoparticles (CuNPs) has been developed using Priestia sp. bacteria utilizing a variety of low-cost agro-industrial wastes and byproducts. The biosynthesis of CuNPs was conducted using glucose medium and copper ions salt solution, then it was replaced by utilizing low-cost agro-industrial wastes. UV–visible spectroscopy, dynamic light scattering (DLS), X-ray diffraction (XRD), High-resolution transmission electron microscope (HR-TEM), Attenuated Total Reflectance and Fourier transform infrared (ATR-FTIR), and zeta potential were used to characterize the biosynthesized CuNPs. The cytotoxicity of CuNPs using Vero -CCL-81 cell lines, and antibacterial and antitumor properties using human colon epithelial colorectal adenocarcinoma Caco-2-HTB-37 cell lines were assessed. The UV–visible and DLS studies revealed CuNPs formation, with a maximum concentration of 6.19 ppm after 48 h, as indicated by a 0.58 Surface plasmon resonance (SPR) within 450 nm and 57.73 nm particle size. The 16S rRNA gene analysis revealed that Priestia sp. isolate is closely related to Priestia megaterium and has been deposited in the NCBI GenBank with accession number AMD 2024. The biosynthesis with various agro-industrial wastes indicated blackstrap sugar cane molasses being the most effective for reducing CuNPs size to 3.12 nm owing to various reducing and stabilizing active compounds. The CuNPs were free of contaminants, with a sphere-shaped structure and a cytotoxicity assessment with an IC50 of 367.27 μg/mL. The antibacterial activity exhibited by the most susceptible bacteria were Bacillus cereus ATCC 11788 and Staphylococcus aureus ATCC 6538 with inhibition zones of 26.0 mm and 28.0 mm, respectively. The antitumor effect showed an IC50 dose of 175.36 μg/mL. Based on the findings, the current work sought to lower product costs and provide a practical solution to the environmental contamination issues brought on by the buildup of agricultural wastes. In addition, the obtained CuNPs could be applied in many fields such as pharmaceuticals, water purification, and agricultural applications as future aspects.

Effects of Curcumin and Doxorubicin on the Viability of Neuroblastoma Cancer Cell Line

Aim: Neuroblastoma has a very important place among childhood diseases, and despite all the methods used in treatment, it is very difficult to prevent neuroblastoma invasion. The number of studies showing that curcumin, the most active component of turmeric, is not toxic, is increasing day by day. In this study, the anticancer activities of curcumin (Cur), one of the important active compounds, were demonstrated in the Neuroblastoma cancer cell line (Na2B). Method: Neuroblastoma cell line was used in the study. To determine the IC50 doses of Dox and Cur, Na2B cells line were cultivated with an automatic pipette. MTT analysis was performed to analyze cell survival (viability). Inhibition levels in the cells were determined at 24 and 48 hours. Results: While the IC50 of Na2B cell proliferation was approximately 124.5 uM at the 48th hour in Dox-treated cells, the IC50 value of Cur at the 48th hour was found to be 224.6 uM. Conclusion: These results showed that Cur could be an alternative agent in the treatment of neuroblastoma, and its fewer side effects compared to other chemotherapeutic agents such as Dox would increase its preferability.

Phytochemical profiling and bioactivity analysis of Citrus japonica leaves volatile oil from Palestine

BackgroundExploring the therapeutic potential of unutilized plant parts from agricultural crops represents a promising strategy for discovering novel medications with high positive economic value. This study aimed to characterize the composition, antidiabetic, anti-obesity, antioxidant, and cytotoxic effects of volatile oil (VO) extracted from the leaves of Citrus japonica trees. This is the first research to assess the C. japonica VO’s anti-obesity, anti-lipase, and cytotoxic properties. Using 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assays and gas chromatography–mass spectrometry (GC–MS) analysis, the components of VO and its capacity to suppress the growth of cancer and other abnormal cells were ascertained, respectively. Stable 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radicals, p-nitrophenyl butyrate (PNPB), and dinitrosalicylic acid (DNSA) assays were employed to determine antioxidant, anti-obesity, and antidiabetic activities, respectively.ResultsThe C. japonica leaf showed significant diversity in type and percentage of VO molecules. Overall, 45 compounds were identified in the VO, constituting 99.69% of the total oil composition. γ-Muurolene (28.12%), β-eudesmol (10.93%), γ-eudesmol (8.44%), germacrene B (7.39%), and elemol (7.27%) are the major characterized molecules. According to the inhibition percentage results of DPPH free radicals, porcine pancreatic lipase, and α-amylase, the VO exhibits strong antioxidant properties and weak inhibitory effects on lipase and α-amylase enzymes. The C. japonica VO showed a moderate cytotoxic effect against Hep3B and considerable activity on B16F1, CaCo-2, HeLa, MCF-7, and HepG2, with IC50 doses in the range of 69.7–171.96 µg/mL. The VO cytotoxic effect IC50 against the normal cell line LX-2 was 224.95 µg/mL.ConclusionThe current study collectively presented the chemical constituents of C. japonica leaf VO from Palestine for the first time and demonstrated its inhibitory effects against DPPH free radicals, porcine pancreatic lipase, and α-amylase. The results suggest that C. japonica leaf VO has the potential to be used as a natural supplement to prevent or treat cancer, as well as in the food industry as a natural antioxidant.Graphical

Abstract C073: FOLFIRINOX with Glycogen Synthase Kinase-3 Beta (GSK-3β) Inhibitor Elraglusib and Transforming Growth Factor- β (TGFβ) Inhibitor Losartan in Untreated Metastatic Pancreatic Ductal Adenocarcinoma(PDAC): Interim analysis of safety cohort

Abstract Introduction: Epithelial-mesenchymal transition (EMT) is one of the key drivers of aggressive biology and development of chemoresistance in PDAC. Inhibition of glycogen synthase kinase 3β (GSK3β) may offer a promising novel approach for modulating dynamic EMT plasticity to overcome resistance to FOLFIRINOX (5-Fluororuacil, Irinotecan, Oxaliplatin, FFX). Methods: Cell line panels comprising both epithelial (E) and quasi-mesenchymal (QM) subtypes were used and treated with IC50 (0.3125 mM) and IC100 (1.25 mM) doses of elraglusib (ELR). Gene expression patters in response to treatment were analyzed using bulk RNA sequencing. 6 patients in the safety run-in cohort of NCT05077800 treated with standard doses of FFX, ELR and losartan were included in this interim analysis. Biopsy specimens were processed for RNA-in situ hybridization (RNA-ISH), a clinically validated assay, and Nanostring (GeoMx) spatial transcriptomics. Results: In preclinical models, exposure to the higher IC100 (1.25 mM) doses of ELR in the QM cell lines caused a shift in transcriptional expression patterns similar to the E cell line, with strong downregulation of EMT. FFX exposure alone in QM cell lines was associated with significant resistance, but addition of ELR (even at IC50 dose) caused a synergistic effect with decreased cell viability. However, in the E cell lines, there was no benefit from addition of ELR. In the safety cohort, clinical results are encouraging with a long-term (>6 month) response in 3/6 patients. 2/6 achieved partial response, 1/6 had stable disease. Nanostring (GeoMx) spatial transcriptomic profiling on available biopsy samples showed significant global gene expression changes before and after treatment. There were concordant changes in 97 differentially expressed genes (DEGs) within the epithelial compartment with treatment, similar to ELR induced changes in PDAC cell lines, with a shift in transcriptional expression patterns from QM to E subtype. Gene set enrichment analysis demonstrated downregulation of the EMT pathway, as well as decreased TGF-β signaling. RNA-ISH analysis on baseline biopsy samples showed that the two partial (best) responders had higher degrees of QM marker expression (82.58% and 63.76%). The regimen was well tolerated with no dose limiting toxicity noted in the safety cohort. Conclusion: Our experiments demonstrate that the E subtype PDAC cell lines are inherently sensitive to FFX chemotherapy with no added advantage with GSK3β inhibition. Meanwhile, GSK3β inhibition in QM cell lines induces mesenchymal to epithelial transition, thereby rendering it sensitive to FFX. The data from our safety cohort provide a clinical proof of concept that GSK-3β inhibition with ELR, in combination with FFX, may be an effective therapeutic strategy in metastatic PDAC. NCT05077800 is currently enrolling. Citation Format: Priyadarshini Pathak, Eric Lin, Ildiko Phillips, David Ting, Colin Weekes. FOLFIRINOX with Glycogen Synthase Kinase-3 Beta (GSK-3β) Inhibitor Elraglusib and Transforming Growth Factor- β (TGFβ) Inhibitor Losartan in Untreated Metastatic Pancreatic Ductal Adenocarcinoma(PDAC): Interim analysis of safety cohort [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Advances in Pancreatic Cancer Research; 2024 Sep 15-18; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(17 Suppl_2):Abstract nr C073.

The Potential Anticancer Potency of Kolaviron on Colorectal Adenocarcinoma (Caco-2) Cells.

Globally, colorectal cancer (CRC) is categorized as the third type of cancer associated with mortalities. Chemotherapeutic drugs such as cisplatin can be used to treat cancer-affected patients. However, several adverse effects are associated with its application. This motivated the researchers to search for alternatives that are more efficient and have fewer undesirable effects. Kolaviron is a bioflavonoid that has been reported to have antioxidant and anti-inflammatory properties. This study aimed to compare the anticancer effects of kolaviron and cisplatin on Caco-2 cells. The IC50 of kolaviron and cisplatin were calculated, and redox status, apoptotic-related proteins and the cell cycle were also examined. Caco-2 cells were treated with kolaviron ( ⅓, and ½ of IC50 dose) and cisplatin (IC50 dose) for 24 h and 48 h. Cell viability was assessed using the MTT protocol. Redox status and apoptotic-related proteins, in addition to the cell cycle, were examined. The MTT assay showed the IC50 of kolaviron is 9.49 μg/mL, and that of cisplatin is 2.71 μg/ml against Caco-2 cells. Further, both doses of kolaviron significantly increased the leakage of lactate dehydrogenase (LDH), the production of reactive oxygen species (ROS), and lipoperoxidation (LPO), besides decreasing the antioxidant potency of tumor cells as revealed by the diminished reduced glutathione (GSH). At the molecular level, a significant increase in the levels of p53, cytochrome c, Bax, and caspase 3 was recorded, coupled with a decrease in the level of Bcl2, after treating the Caco-2 cells with kolaviron and cisplatin. Furthermore, kolaviron demonstrated asserted more effects on apoptosis and increased cell percentage in the subG1 phase. In addition, a notable decrease in the expression of proliferating cell nuclear antigen (PCNA) and cyclin D1 is associated with an increase in the expression of tumor protein P53 (TP53) in kolaviron-treated Caco-2 cells cancerous cells. Conclusively, these data suggest that kolaviron has a potential antitumor capacity against colorectal cancer via multiple pathways, including enhancement of ROS production, redox status, p53 pathway, and apoptosis. Therefore, this study authenticated the capability of kolaviron as a valuable chemotherapeutic agent.

Comparison of Anticancer, Antioxidant, Enzyme Inhibitory Effects and Phytochemical Contents Between Edible Lettuce (Lactuca sativa) and a New Wild Species (Lactuca anatolica).

In this study, the bioactive components, enzyme inhibitory, antioxidant and anticancer potentials of edible (L. sativa) and a new species (L. anatolica) of Lactuca were evaluated and compared. The quantitative analyzes of the bioactive components of L. sativa (LS) and L. anatolica (LA) were analyzed quantitatively by GC-MS and Orbitrab HPLC-HRMS. Antioxidant, enzyme inhibitory and anticancer properties were analyzed by various assays. In general, LA exhibited more stronger antioxidant properties compared to LS. The extracts showed similar inhibitory effects on these enzymes. It was determined that LS was dominant in terms of linoleic acid (23.71 %), while LA contained a high level of α-linolenic acid (31.70 %). LA and LS inhibited the viability of A549 and MCF-7 cells in a dose-dependent manner. IC50 values for LA, LS and cisplatin were determined as 120.3, 197.5, 4.3 μg/mL in A549 cell line and 286.2, 472.8, 7.2 μg/mL in MCF-7 cell line, respectively. It was revealed that LA and LS treatment at 50 μg/mL concentrations in A549 cells completely suppressed the colony forming capacity, and treatment with IC50 doses inhibited cell migration, and triggered apoptosis by regulating caspase-3, cPARP, p53 and p21. The findings of this study suggested that these species have significant pharmacological potential.

Insights into free radicals scavenging, α-Amylase inhibition, cytotoxic and antifibrotic activities unveiled by Peganum harmala extracts

BackgroundPeganum harmala L. is used in traditional medicine to treat several health ailments. Hence, the present work aimed to investigate the DPPH free radical scavenging, α-amylase, cytotoxic, and antifibrotic effects of the hydrophilic extract and fixed oil obtained from P. harmala seeds.MethodsThe hydrophilic extract and fixed oil of P. harmala were assessed for their abilities to scavenge DPPH free radicals and inhibit α-amylase using reference bioassays. The cytotoxicity was assessed on several cancer and normal cell lines, including B16F1, Caco-2, COLO205, HeLa, Hep 3B and Hep G2, MCF-7, and HEK-293 T cells. The MTS assay was used to evaluate the antifibrotic capabilities utilizing the human hepatic stellate (LX-2) cell line.ResultsP. harmala plant fixed oil has potent DPPH free radical scavenging activity with an IC50 dose of 79.43 ± 0.08 µg/ml. Besides, the hydrophilic extract has a poor anti-α-amylase effect compared with the antidiabetic drug Acarbose, with IC50 doses of 398 ± 0.59 and 25.11 ± 1.22 µg/ml, respectively. In addition, the growth of MCF-7, Hep3B, HepG2, HeLa, COLO205, CaCo2, B16F1, and HeK293t was inhibited by P. harmala hydrophilic extract with IC50 doses of 121.34 ± 1.71, 268.3 ± 0.75, 297.20 ± 1.00, 155.60 ± 1.14, 150.01 ± 0.51, 308.35 ± 0.53, 597.93 ± 1.36, and 5.38 ± 0.99 µg/ml, respectively. In addition, at 1000 µg/ml, 5-Fluorouracil reduced fibrosis cells by 0.089%, while the hydrophilic extract decreased the number of LX-2 cells by 5.81%.ConclusionP. harmala plant-fixed oil exhibits potential antioxidant properties. While the hydrophilic extract showed limited effectiveness as an anti-α-amylase agent and demonstrated notable cytotoxic effects against various tested cancer cell lines. Furthermore, this extract significantly reduces the number of LX-2 fibrotic cells. These findings emphasize the therapeutic potential of these products in managing various health disorders and warrant further investigation into their mechanisms of action and clinical applications.

Evaluation of Boric Acid Treatment on microRNA-127-5p and Metastasis Genes Orchestration of Breast Cancer Stem Cells.

Coregulation of microRNAs (miRNAs) and cancer stem cells (CSCs) is very important in carcinogenesis. miR-127-5p is known to be downregulated in breast cancer. In this study, we aimed to investigate how boric acid (BA), known for its previously unstudied anti-cancer properties, would affect the expression of miR127-5p and genes responsible for breast cancer stem cells (BC-SCs) metastasis. BC-SCs were isolated from human breast cancer cells (MCF-7) by immunomagnetic cell separation and characterized with flow cytometry and sphere formation. The viability of BC-SCs and the determination of its IC50 value in response to boric acid (BA) were assessed via the MTT assay. Boric acid exhibited dose- and time-dependent inhibition of cell viability in cells. The IC50 doses of boric acid in MCF-7 cells and BC-SCs were 45.69mM and 41.27mM, respectively. The impact of BA on the expression of metastatic genes and miR127-5p was elucidated through RT-qPCR analysis. While the expression of the COL1A1 (p < 0.05) and VIM (p < 0.01) was downregulated, the expression of the miR-127-5p, ZEB1 (p < 0.01), CDH1 (p < 0.05), ITGB1 (p < 0.05), ITGA5 (p < 0.05), LAMA5 (p < 0.01), and SNAIL (p < 0.05), was up-regulated in dose-treated BC-SCs (p < 0.001) to the RT-qPCR results. Our findings suggest that boric acid could induce miR-127-5p expression. However, it cannot be said that it improves the metastasis properties of breast cancer stem cells.

Biogenic synthesis of silica nanoparticles from industrial hemp waste for sustainable applications: Characterization and potential environmental benefits

SiO2, commonly referred to as silica, represents a versatile material utilized across various domains including biomedical, construction, food, energy, and cosmetic sectors. This study is anticipated to offer insights into the synthesis of SiO2-NP from industrial hemp waste, thus contributing to sustainability efforts and waste management systems. Within this investigation, SiO2-NP were biogenically synthesized utilizing waste leaves from industrial hemp, a sustainable plant source. The biogenically synthesized SiO2-NP underwent characterization via several techniques including UV–visible absorption spectroscopy (UV–Vis), X-ray diffraction (XRD), Energy Dispersion X-ray Spectrometer (EDX), and Scanning Electron Microscopy (SEM) analysis. The UV–Vis spectrum indicated an absorption peak of 360 nm for SiO2-NP. XRD analysis determined an average particle size of 28 nm. SEM analysis revealed spherical morphology of the synthesized SiO2-NP. Additionally, EDX analysis unveiled a composition consisting of 50.6 % O and 27.2 % Si. SiO2-NP exhibited antibacterial activity against both gram-positive and gram-negative bacteria strains. According to anticancer activity analyses, IC50 doses of SiO2-NP in MCF7 and MDA-MB-231 breast cancer cells were found to be 146.73 ± 7.54 and 54.93 ± 5.54, respectively, for 48 h. After treating the cells with these doses, an increase in the expression of apoptosis-related genes was detected. The SiO2-NP has an IC50 value with 2.69 ± 0.42 mg/mL against AChE enzyme activity.

Natural products to enhance anti-tumor effects of trastuzumab and paclitaxel on the SKBR3, which is human breast cancer cell line.

e13051 Background: Neoadjuvant chemotherapy (ChT) with targeted therapy is the standard treatment for human epidermal growth factor 2 (HER2)- positive breast cancer (BC). In this context, Trastuzumab (Tz) and ChT significantly improved the pathological complete response (pCR) rate. Tz and Paclitaxel (PTX) combination therapy is also one of the neoadjuvant treatments used in HER2+BC, and despite the effectiveness of this treatment, there is a critical need for innovative approaches and further research that will improve the neoadjuvant response rate. Addressing this, the NaturaProDB tool presents a novel avenue for predicting in silico natural non-toxic compounds with potential anti-cancer properties. Our study aims to investigate synergistic effect of natural products proposed by this database on theurapeutic action of Tz and PTX on BC. Methods: The NaturaProDB database was searched using the ‘Target Cancer’ option in order to identify those natural products with anti-BC potential and the corresponding experimentally verified target genes. Using this database, Biochanin A (BCA) and Delphinidin (Dp) were selected as natural compounds because they both commonly inhibit the MAPK signaling pathway. We used the HER2+ SKBR3 cell line to determine IC20, IC30 and IC50 doses for BCA, Dp, Tz, and PTX through MTT assay. Notably, all experiments were done after 72 hrs treatment of all compounds. Further, various combinations within different concentrations of natural products and Tz and PTX were tested to define additive anti-proliferative effects on SKBR3 cell line. Results: We first defined two natural products with drug-like properties, BCA and Dp, which have anti-proliferative effect on BC. Treatment of SKBR3 with increasing concentrations of BCA and Dp caused dose-dependent proliferation inhibition and IC50 dose of compounds were defined as 42,70uM and 40,70uM, respectively. In addition, IC20 dose of Tz was 2,8µM, while IC30 dose of PTX was 2,3nM.Then, we showed that combinations of increasing doses of Dp (from 30µM to 90 µM) with IC20 Tz caused significant dose-dependent cell death compared to the control group (p&lt;0.0001). And the combinations of increasing doses of BCA (from 45µM to 360 µM) with IC20 Tz led similar significant results (p&lt;0.0001). Conclusions: Herein, it is shown that combining natural compounds (Dp, BCA) with conventional treatments (PTX,Tz) predicted for the SKBR3 cell line can enhance the inhibitory effect on cell proliferation. This study is one of the rare first cell line studies in which the synergistic effect of BCA and Dp, which are natural compounds selected using the NaturaProDB database, were combined with ChT and Tz to show their synergistic effect, and these findings indicate that synergistic combination treatments acting through different pathways (MAPK signaling pathway) can be used in the treatment of HER2+ and TN BC. It opens avenues for further research.

A sustainable green-approach for biofabrication of chitosan nanoparticles, optimization, characterization, its antifungal activity against phytopathogenic Fusarium culmorum and antitumor activity.

Chitosan is a natural non-toxic, biocompatible, biodegradable, and mucoadhesive polymer. It also has a broad spectrum of applications such as agriculture, medical fields, cosmetics and food industries. In this investigation, chitosan nanoparticles were produced by an aqueous extract of Cympopogon citratus leaves as a reducing agent. According to the SEM and TEM micrographs, CNPs had a spherical shape, and size ranging from 8.08 to 12.01 nm. CNPs have a positively charged surface with a Zeta potential of + 26mV. The crystalline feature of CNPs is determined by X-ray diffraction. There are many functional groups, including C꞊C, CH2-OH, C-O, C-S, N-H, CN, CH and OH were detected by FTIR analysis. As shown by the thermogravimetric study, CNPs have a high thermal stability. For the optimization of the green synthesis of CNPs, a Face centered central composite design (FCCCD) with 30 trials was used. The maximum yield of CNPs (13.99 mg CNPs/mL) was produced with chitosan concentration 1.5%, pH 4.5 at 40°C, and incubation period of 30 min. The antifungal activity of CNPs was evaluated against phytopathogenic fungus; Fusarium culmorum. A 100% rate of mycelial growth inhibition was gained by the application of 20mg CNPs/mL. The antitumor activity of the green synthesized CNPs was examined using 6 different cell lines, the viability of the cells reduced when the concentration of green synthesized CNPs increased, the IC50 dose of the green synthesized CNPs on the examined cell lines HePG-2, MCF-7, HCT-116, PC-3, Hela and WI-38 was 36.25 ± 2.3, 31.21 ± 2.2, 67.45 ± 3.5, 56.30 ± 3.3, 44.62 ± 2.6 and 74.90 ± 3.8; respectively.

Imidazole-Derived Alkyl and Aryl Ethers: Synthesis, Characterization, In Vitro Anticancer and Antioxidant Activities, Carbonic Anhydrase I-II Inhibition Properties, and In Silico Studies.

Imidazole derivatives display extensive applications in pharmaceutical chemistry and have been investigated as bioactive compounds for medicinal chemistry. In this study, besides the starting materials (3a-c and 4a-c), synthesis, characterization, and biological activity studies were conducted on a total of 18 compounds, nine of which are known and the other nine are original. The compounds investigated in the study are a series of alkyl (7-15) and aryl (16-24) ether derivatives bearing substituted phenyl and imidazole rings, which were characterized using various methods including 1H NMR, 13C NMR, FT-IR analysis, elemental analysis, and mass spectroscopy. Computer-aided drug design studies have been carried out to predict the biological activities of compounds. Besides DFT calculations, the binding affinities of the compounds to EGFR, VEGFR2, FGFR1, HSP90, hCA I, and hCA II were investigated. Additionally, drug-likeness and ADME analyses were performed on the compounds. Anticancer, antioxidant, and enzyme inhibition activity tests were performed in biological activity studies on the synthesized compounds. Among the synthesized compounds, compounds 17 and 19-24 generally exhibited inhibition profiles against the widespread cytosolic hCA I isozyme with IC50 values ranging from 4.13 to 15.67 nM and cytosolic hCA II isozyme with IC50 values ranging from 5.65 to 14.84 nM. L929 (mouse fibroblast cell line) was used as the control healthy cell line, and MCF7 (breast cancer), C6 (rat glioblastoma), and HT-29 (colon cancer) cells were used in cell culture studies as cancer cell lines. Before the study on cancer cells, all compounds were examined on healthy cells, and their cytotoxicity was determined. As a result of these data, studies continued with six compounds determined to be nontoxic. On cancerous cells, it was determined that compounds 3a, 3b, 4a, 4b, 4c, and 7 had cytotoxic effects on both colon cancer and brain tumors. It was found that compound 3b had a more toxic effect than cisplatin on the glioma cell line with an IC50 value of 10.721 ± 0.38 μM, and compound 3a had a more toxic effect on the colon cancer cell line with an IC50 value of 20.88 ± 1.02 μM. However, it was determined that the same compounds did not have a statistically significant effect on breast cancer. Flow cytometry studies also showed that when the IC50 dose of compound 3b was applied to the C6 cell line, the cells tended to early and late apoptosis. Additionally, it has been shown by flow cytometry that the cell cycle stops in the G0/G1 phase. A similar effect was observed in the colon cancer cell line with compound 3a. Compound 3b caused early and late apoptosis of the colon cancer cell line with the applied IC50 dose and stopped the cell cycle in the G0/G1 phase. Finally, the FRAP method studied all synthesized compounds' antioxidant effects. According to the measured antioxidant power results, it was determined that no compound had a more effective reducing power than vitamin E.

Modulation of IKK/NF-κB Signaling: A Therapeutic Mechanism of Shilajit in Breast Cancer Cells

Background: Shilajit is a natural pale-brown to blackish-brown phytocomplex that exudes from mountain rocks in various parts of the world. In recent years, numerous pharmacological effects of shilajit have been explored through extensive experiments. Shilajit is known as an antioxidant and anti-inflammatory agent. Activation of the nuclear factor kappa B (NF-κB) signaling pathway could be an important player in inflammation-driven tumor progression. Therefore, the current study evaluated the anti-inflammatory effect of shilajit on a breast cancer cell line (MCF7). Methods: In this experimental study, the MCF7 cell line was treated with shilajit, and an MTT assay was applied to analyze the half-maximal inhibitory concentration (IC50) after 72 hours of treatment. Following this, the apoptotic rate was assessed using flow cytometry, and p50, RelB, and IKKα/β gene expression were evaluated using real-time PCR assay. Results: Shilajit had potent cytotoxic activity in a dose- and time-dependent manner with an IC50 of 280 μg/mL. Based on the Annexin-PI analysis, the IC50 concentration of this compound induced significant apoptosis in the cells, possibly through suppression of NF-κB (p50, RelB, and IKKα/β)-regulated genes. The real-time PCR results indicated that treating MCF7 cells at the IC50 dose of shilajit for 72 hours could reduce the mRNA expression levels of p50, RelB, and IKKα/β in the cells. Conclusion: The findings of the present experiment showed that shilajit may have promising anti-breast-tumor and anti-inflammatory activity through inhibition of the NF-κB signaling pathway. Therefore, shilajit can help treat breast cancer in combination with other standard treatments.

Rhein inhibits cell proliferation of glioblastoma multiforme cells by regulating the tgfß and apoptotic signaling pathways

Rhein (4,5-dihydroxyanthraquinone-2-carboxylic acid) is a plant metabolite found in rhubarbs. It inhibits cell proliferation and stimulates apoptosis in in vivo and in vitro. However, research into the molecular mechanisms of action is insufficient for recommending it as a therapeutic agent. Therefore, this study aims to investigate the antiproliferative, apoptotic, and antimetastatic effects of rhein by targeting the TGF-β signaling pathway, and apoptotic pathway in glioblastoma cells (U87 GBM). In this study, the XTT assay was utilized to determine cell viability, the colony formation assay to measure cell survival and proliferation, RT-qPCR for the analysis of gene expressions, and ELISA for the detection of proteins. U87 GBM cells were treated with varying concentrations of rhein (5-100 µM) in a time-dependent manner (24, 48 h), after which the percentage of cell viability was calculated. The colony formation assay was performed by treating cells with the IC50 dose of rhein. According to the XTT assay, the IC50 dose of rhein was determined as 10 µM at 24 h. The ability to form colonies was significantly decreased in the cells of the treatment group. According to the gene expression analysis, rhein increased the mRNA levels of CASP3, -8, -9, BAX, and TGF-β1 genes, while a notable decrease was observed in the BCL-2, SMAD2, SMAD3, and TIMP1 genes. In conclusion, it was determined that rhein induces apoptosis via the non-canonical TGF-β pathway.

Abstract LB027: Identifying natural inhibitors of androgen receptor (AR) and its variants from NCI Natural Product Repository

Abstract Second-generation androgen ablation therapies (ADT), enzalutamide, and abiraterone are initially effective on metastatic castration-resistant prostate cancer (mCRPC). However, nearly 30-40% of patients eventually become resistant to these treatments due to the presence of androgen-receptor variants (AR-V), which lack a ligand-binding domain. The grim reality that metastatic CRPC is virtually incurable for most patients poses a significant challenge for clinicians striving to enhance these individuals' quality of life and lifespan. Considering drug discovery efforts, we partnered with the National Cancer Institute's (NCI) Natural Product Repository, which possesses a diverse collection of natural product extracts derived from plant, marine, and microbial organisms. We developed high-throughput phenotypic screening assays to screen the crude natural product extract libraries on AR and AR-variants CRPC cell lines (C4-2B and 22Rv1). Several extracts inhibited the growth of both CRPC cell lines and were further confirmed for dose-dependent cell viability inhibition assays. Once we determined the IC50 dose, we performed a Western blot analysis that confirmed the inhibition of AR and AR-V7 expression in CRPC cells.In subsequent studies, we narrowed down by fractionating the most potent extracts and confirmed their ability to inhibit AR expression and signaling on CRPC cell lines. Currently, we are characterizing the potent compounds and their inhibitory activity on AR and AR-variants, in vitro and in vivo CRPC models. Many nutraceuticals have been shown to target AR signaling, synthesis, or degradation which effectively inhibited the growth of CRPC. Our collaborative efforts with NCI will permit us find N-terminal AR degrading molecules (AF-1), which are essential for the AR transcriptional activities of CRPC. In sum, discovery of ovel small molecules from our drug development program have the potential to translate these effective natural products into drugs with clinical benefits to treat CRPC. Citation Format: Neha Tyagi, Sepideh Hosseiniporgham, Ashish Tyagi, Balaji Chandrasekaran, Katherine Bonilla, Inna Kriger, James Saccchettini, Chendil Damodaran. Identifying natural inhibitors of androgen receptor (AR) and its variants from NCI Natural Product Repository [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr LB027.