Cancer Cell Caco-2 Research Articles

Anticarcinogenic cationic peptides derived from tryptic hydrolysis of β-lactoglobulin.

This study investigated the tryptic hydrolysis of β-lactoglobulin (BLG) for 30, 60, 90, and 120 min at 1/200 E/S (enzyme/substrate ratio, w/w) to prepare potentially anticarcinogenic peptides. The properties of hydrolysates were characterized, including degree of hydrolysis, free amino acids, SDS-PAGE, FTIR, and antioxidant activity employing DPPH-assay, β-carotene/linoleic acid, and FRAP assay. BLG tryptic hydrolysate produced after 60 min hydrolysis recorded the highest antioxidant activity, and LCMS analysis revealed 162 peptides of molecular masses ranging from 800 to 5671Da, most of them are of hydrophobic nature. Within the low-MW peptide group (24 peptides), there were nine hydrophobic basic (HB) and seven hydrophobic acidic (HA), representing 38% and 29%, respectively. The HB peptides may be responsible for the considerable biological activity of the hydrolysate. With dominant basic character supporting the carcinogenic activity of this hydrolysate. The in vitro anticancer activity against Mcf-7, Caco-2, and A-549 human cancer cell lines proliferation by MTT assay recorded IC50% at 42.8, 76.92, and 45.93 μg/mL, respectively. Treating each cell line with IC50% of the hydrolysate for 24 h increased the apoptosis by enhancing the expression of caspase-9 by 5.66, 7.97, and 3.28 folds over the untreated control and inhibited angiogenesis by reducing VEGFR-2 expression by about 56, 76, and 70%, respectively, indicating strong anticancer and antiangiogenic actions on human cancer cells. BLG tryptic hydrolysate may serve as a natural anticarcinogenic agent. The results of this study demonstrated that BLG hydrolysates have direct anticancer and antiangiogenic effects on human cancer cells. The chemical composition and characteristics of the BLG tryptic hydrolysate influence these biological and anticancer activities. The tryptic hydrolysates were generally effective against the three cancer cell lines studied (Mcf-7, Caco-2, and A-549). This effectiveness was assessed by measuring cell proliferation using the MTT assay and by evaluating their impact on angiogenesis through inhibition of VEGFR-2 activity. Future studies may focus on enhancing the anticarcinogenic effectiveness of the peptides by isolating and evaluating the most prominent individual peptide and varying the treatment conditions.

MiRNA-sequencing and bioinformatics analyses of key miRNAs and their associated genes regulated by EC-synthetic retinoids treatment in Caco-2 cancer cells

miRNA-sequencing and bioinformatics analyses of key miRNAs and their associated genes regulated by EC-synthetic retinoids treatment in Caco-2 cancer cells

Icariside II relieves radiation enteritis by regulating PINK/Parkin-mediated mitophagy.

Radiation enteritis (RE) is one of the major side effects of radiotherapy. So far, there are no effective drugs for preventing the disease process. Icariside II (ICS II) is a highly efficient monomer compound extracted and purified from the classic Chinese medicinal herb Epimedium. It has anti-inflammatory, antioxidant, and immunomodulatory effects. However, the role and mechanism of ICS II on radiation enteritis are not clear. Here, we reveal the role of ICS II in radiation enteritis by using an irradiation-induced rat model and a human colorectal cancer cell (CaCo2). After intragastric administration, HE staining and Tunel staining to observe the histopathological changes in the colon, and TEM to observe the ultrastructure of mitochondria; The antioxidant indexes and mitochondrial function-related markers of colon tissues were determined; DCFH-DA fluorescent probe were used to detect the cellular ROS level, JC-1 staining was used to detect the changes in mitochondrial membrane potential, and Western Blot was used to detect related protein expression. The results showed that ICS II could reduce intestinal injury and attenuate the radiation-induced oxidative stress and inflammatory response. In addition, ICS II could effectively attenuate mitochondrial damage and activate mitochondrial autophagy in rats. Mechanistically, ICS II activates mitochondrial autophagy-related protein expression to rescue radiation-induced damage to mitophagy. We found that by inhibiting mitophagy, the therapeutic effect of ICS II can be eliminated and our data suggest that ICS II may be a new and effective drug candidate for the treatment of radiation enteritis.

Chan Seeds (Hyptis suaveolens L.) Storage Proteins: Isolation, Characterization, and Cytotoxic Effect on Three Cancer Cell Lines

Since pre-Hispanic times, chan seeds (Hyptis suaveolens L.) have been used as food and in traditional medicine. However, there is still a lack of knowledge about the antioxidant capacity and cytotoxic effect of their storage proteins on cancer cells. Thus, these were investigated in this study. The total protein content of the seeds was 19.5% (dry base), and its protein fractions were confirmed via Tris-Tricine-SDS-PAGE electrophoresis as 43.1% glutelins, 30.9% albumins, 23% globulins, and 2.9% prolamins. The antioxidant capacity determined by ABTS showed the highest percentage of inhibition for the prolamins fraction of 23.6% (half-maximal inhibitory concentration [IC50]: 1.38 µg protein/mL) and the lowest percentage for the glutelins fraction of 6.3% (IC50: 4.51 µg protein/mL). The cytotoxic activity against the murine lymphoma L5178Y, human cervical (Hela), and colorectal (Caco-2) cell lines revealed that prolamin exerted superior inhibition on the Hela and Caco-2 cancer cells, with IC50 values of 0.49 and 0.44 mg protein/mL, respectively. This study underscores the potential of chan seed proteins as natural antioxidants and anticancer agents.

Berries, Leaves, and Flowers of Six Hawthorn Species (Crataegus L.) as a Source of Compounds with Nutraceutical Potential.

Designing new forms of food, food additives, and nutraceuticals is necessary due to the growing needs of consumers, as well as the inflammation of civilization diseases, the prevention and treatment of which can be significantly supported by dietary intervention. For this reason, this study aimed to obtain highly bioactive preparations in the form of powders from the fruits, leaves, and flowers of six species of hawthorn (Crataegus L.) using solid phase extraction (SPE). Ultra-performance liquid chromatography analysis (UPLC-PDA-MS/MS) showed a high concentration of phenolic compounds (in the range from 31.50 to 66.06 mg/g), including the highest concentration in hawthorn fruit preparations. Fruit preparations also showed the highest antioxidant activity (through scavenging of O2˙- and OH˙ radicals), antidiabetic activity (inhibition of α-amylase and α-glucosidase), and anticancer activity, mainly against colon cancer cells (Caco-2). At the same time, hawthorn flower preparations showed the highest biocompatibility against normal colon cells (CCD841CoN) and anti-inflammatory activity (trypsin inhibition). Correlation and principal component analysis (PCA) showed that the health-promoting potential was most influenced by the content of falavan-3-ols. The above findings provide a basis for the industrial use of the developed preparations, which is in line with the current trend in food technology related to the search for new sources of bioactive compounds and the design of highly bioactive food.

Green-synthesized silver nanoparticles induced apoptotic cell death in CACO2 cancer cells by activating MLH1 gene expression

MLH1 (Mult homolog1) gene is the main element of Multlα heterodimer and plays a role in the repair of base-base mismatches and deletion and addition loops. When the MLH1 protein is not present, the number of errors that remain unrepaired increases, and this can lead to the formation of tumors in the body. Colorectal cancer is the third most common type of cancer in terms of incidence and the third type of cancer in terms of mortality worldwide. In this regard, the present study was conducted with the aim of investigating the effect of biological silver nanoparticles with anticancer properties and MLH1 gene expression on cancer cell samples of people with colorectal cancer. In this study, the green synthesis of silver nanoparticles was carried out by precipitation method with reduction of silver ions by leaf and flower extract of Moringa oleifera plant. Then, silver nanoparticles were confirmed using UV-visible spectroscopy, transmission and scanning electron microscopy. The cytotoxic effects of nanoparticles on cells were evaluated by MTT colorimetric method within 42 h. After RNA extraction of treated cells, cDNA synthesis and primers were designed by the Exon-Exon Junction method and the Real-time Polymerase test for MLH1 and Beta-actin genes was repeated three times. The final analysis of the results was done using Graphpad Prism and Rest 2009 software. The presence of a peak at the wavelength of 234 nm for the synthesized silver nanoparticles was confirmed by ultraviolet-visible spectroscopic analysis. The morphological study on the size and shape of the silver nanoparticles showed that the nanoparticles are spherical and have a size between 40 and 34 nanometers in diameter. The leaf extract typically produces smaller, more uniform particles than the flower extract. The Green-Synthesized Silver Nanoparticles of leaf extract were used to evaluation of induced Apoptotic Cell Death in CACO2 Cancer Cells by Activating MLH1 Gene Expression. MTT results showed that the anti-proliferative effect of nanoparticles depends on the concentration of synthesized silver nanoparticles. The treatment of MLH1 cancer cell line and normal with synthesized nanoparticles at a concentration of 0.44 micrograms per ml for 42 h showed the effects of cell toxicity. The percentage of cancer cell death under the influence of quercetin present in Moringa green bio-particles depends on the concentration and time, and this difference is statistically significant compared to the control group (P < 0.05). So that the lethal dose of the extract for 50% survival IC50 in a period of 48 h for intestinal cancer cells was equal to 21 μg/ml, and the expression ratio of the MLH1 gene in the tumor tissue to the adjacent healthy tissue has increased (P ≤ 0.05).

Phosphazene Tripeptide Conjugates: Design, Synthesis, In Vitro Cytotoxicity and Genotoxicity, Molecular Interactions in Binding Pockets on Human Breast and Colon Cancer Cell Lines.

The biological activity of both cyclophosphazenes and peptides makes these compounds important for new studies in medicinal chemistry. For this purpose, five different phosphazene-peptide conjugates synthesized from dichlorocyclotriphosphazene and tyrosine-containing tripeptides. The synthesized compounds were evaluated for their in vitro cytotoxic activities against human breast (MCF-7) and colon (Caco-2) cancer cell lines using MTT assay. The derivatives induced cell death through DNA damage, with notable effects in Caco-2 cell lines. Specifically, DTVV, DTVG, and DTVA were cytotoxic at 50 and 100 μM, while DTVP and DTVM were effective at 25, 50, and 100 μM. DTVM outperformed Tamoxifen at 50 μM in the MCF-7 cell line. DNA damage studies of the compounds were performed using the comet assay method, evaluating tail length, tail density, olive tail moment, head length, and head density parameters. The findings indicated that cell death occurred via a DNA damage mechanism. The molecular intricacies of DTVA, DTVG, DTVM, DTVP and DTVV within the VEGFR2 kinase domain (3VHE) and Cyclophilin_CeCYP16-Like Domain (2HQ6) binding pockets and various interactions, docking scores and potential activities of these derivatives were investigated. The differences in docking scores and interaction profiles highlight the potential efficacy and specificity of these compounds in targeting breast and colon cancer cells. These findings highlight the potential of phosphazene-peptide derivatives as therapeutic agents in cancer treatment.

Facile synthesis of corticiolic acid-a bioactive pharmacophore from natural sources.

Fungal strains have inspired us to find the untapped sources of secondary metabolites. Corticiolic acid (CA, 2,4-dihydroxy-6-pentadecylbenzoic acid; from fungus, Hapalopilus mutans) is one of the core active scaffolds in natural compounds such as Aquastatin-A, B, & C. CA can also be isolated from the plant Lysimachia japonica. CA is a selective inhibitor of PTB1B, a crucial biomarker for anti-diabetic activity. Herein, we report the total synthesis of corticiolic acid achieved via the 9-BBN-based reductive Suzuki-Miyaura coupling of aryl bromide and pentadecane, a key reaction in this strategy. Further, this approach has been explored for the protection-free synthesis of corticiolic acid. The improved synthesis is short, requires mild reaction conditions, and avoids the use of hydrogenation and pyrophoric reagents. Further, the reaction is scalable and does not require protection-deprotection steps. Preliminary studies on cancer cells indicated that corticiolic acid and cordol significantly inhibited the proliferation of HepG2, N2A, and CaCo-2 cancer cells.

Spontaneous gradient copolymers of N-vinylsuccinimide/N-vinylsuccinamic acid with O-cholesteryl (meth)acrylate via RAFT polymerization as potential drug delivery systems

Today, biocompatible and bioinspired well-defined copolymers with the ability to form nanoparticles are of great interest as potential drug delivery systems. In this study, we report the synthesis of novel biocompatible copolymers from monomers with different activity and capable of forming gradient copolymers by RAFT polymerization. In particular, the copolymerization of N-vinylsuccinimide (VSI) and O-cholesteryl (meth)acrylate (Ch(M)A) mediated by S,S’-dibenzyl trithiocarbonate (DBTTC) has been thoroughly studied by varying the monomer ratio and the ratio of monomer to RAFT agent. Important dependencies such as molecular weight and monomer conversion versus time, and molar fraction of monomer units as a function of monomer conversion were investigated. The obtained copolymers were thoroughly characterized using a number of physicochemical methods such as 1H NMR, 1H–13C HSQC and ATR-IR-spectroscopy, size-exclusion chromatography, static and dynamic light-scattering, as well as thermogravimetric analysis. In addition, the reactivity ratios of VSI and ChMA were determined and the dyad and triad compositions of the copolymers were calculated from the obtained values. The synthesized P(VSI-co-Ch(M)A) were subjected to selective hydrolysis of succinimide ring to convert it into succinamic acid. This approach yields a set of bioinspired amphiphilic copolymers based on N-vinylsuccinamic acid (VSAA) and Ch(M)A. The synthesized series of P(VSI-co-Ch(M)A) and P(VSAA-co-Ch(M)A) were used to obtain nanoparticles by nanoprecipitation or self-assembly via direct dissolution in aqueous medium. In addition, the method of surface hydrolysis of VSI units in pre-formed P(VSI-co-Ch(M)A) nanoparticles was applied to produce nanoparticles with hydrophilic negatively charged surface and enhanced stability. All techniques were optimized to prepare nanoparticles with characteristics suitable for systems considered for drug delivery. Successful loading of the antitumor drug irinotecan into nanoparticles was achieved with high encapsulation efficacies. The storage stability of empty and irinotecan loaded nanoparticles were studied in various media (water, saline solution, serum containing cell culture medium) under room and refrigerator conditions. The developed empty nanoparticles exhibited low rate of uptake by macrophages and low cytotoxicity to irinotecan-sensitive colon cancer cells (Caco-2). In turn, the irinotecan-loaded nanoparticles demonstrated inhibitory activity against Caco-2 cells comparable to the free drug.

Terminalia ivorensis demonstrates antioxidant properties and alters proliferation, genomic instability and migration of human colon cancer cells in vitro.

Colorectal cancer is a global killer that causes approximately 940 thousand deaths annually. Terminalia ivorensis (TI) is a tropical tree, the bark of which is used in African traditional medicine for the treatment of diabetes, malaria and ulcer. This study investigated TI as a potential anticancer agent in human colon cells in vitro. TI was extracted sequentially with petroleum ether, chloroform, ethyl acetate and ethanol. Antioxidant activity was assessed by DPPH and FRAP, and differential effects on cell viability, growth, DNA damage, DNA repair, and migration were measured in human colon cancer cells (CaCo-2) and/or non-cancerous human colonocytes (NCM460). The TI phytochemicals most strongly associated with these effects were identified by partial least-squares discriminant analysis. DPPH and FRAP activity were highest in TI ethyl acetate and ethanol extracts (p=0.001). All TI extracts significantly inhibited cell viability and growth and induced DNA damage and inhibited DNA repair in both cell models. The majority of TI extracts were significantly (p=0.01) more toxic to cancer cells than non-cancerous colonocytes. DNA repair was significantly (p=0.001) inhibited in CaCo-2 cells by ethyl acetate extract compared with NCM460 cells. Migration was also significantly inhibited (p<0.001) in CaCo-2 by ethyl acetate (80%) and ethanol extracts (75%). Specific benzoic acids, flavonoids and phenols were identified to be strongly associated with these effects. TI displayed strong antioxidant activity and specific anticancer effects by inducing cell death and DNA damage, and by inhibiting DNA repair, cell proliferation and migration.

Chemical composition, cytotoxic potential and antioxidant properties of Punica granatum peel extract

Purpose: To evaluate the antioxidant activity of pomegranate peel extract and its effects on the survival of colorectal cancer cells (Caco-2) and kidney cells (HEK293). Methods: An ethanol extract was prepared from the outer layer of pomegranate peels. The chemical composition of the extract was examined using high-performance liquid chromatography with diodearray detection (HPLC-DAD) and liquid chromatography with tandem mass spectrometry (LC-MS-MS). Antioxidant activity was determined using DPPH (2,2-diphenyl-1-picrylhydrazyl) and ABTS (2,2'- azinobis(3-ethylbenzothiazoline-6-sulfonic acid)) assays. Cytotoxicity was tested against Caco-2 colorectal cancer cells and HEK293 normal kidney cells using MTT assay. Results: The pomegranate peel extract was rich in phenolic compounds and demonstrated significant antioxidant activity, with IC50 values comparable to Trolox in DPPH and ABTS assays. The extract exhibited selective and concentration-dependent cytotoxicity against Caco-2 cells, with an EC50 of 121.05 ± 13.12 µg/mL, while showing minimal toxicity towards HEK293 cells. Conclusion: Pomegranate peel extract's high phenolic content and potent antioxidant properties highlight its potential as a therapeutic agent against colorectal cancer. The findings suggest that this extract could serve as a viable, natural alternative with reduced toxicity for colorectal cancer treatment.

Synthesis, Crystal Structure, DFT Calculations, Bioactivity Study, and Docking Results of Bis‐Hydrazone Based on 1,2,4‐Triazol‐3‐Thione

AbstractThe new, unexpected bioactive bis‐hydrazone derivative (4) was obtained, in 74 % yield, by reacting two molar equivalents of pyrazole‐4‐carbaldehyde (1) with one molar equivalent of 2,5‐dihydrazineyl‐1,3,4‐thiadiazole (2). The compound was comprehensively characterized, including X‐ray single crystal, DFT calculations, and bioactivity assessments. Hirschfeld surface analysis confirmed the presence of hydrogen bonding interactions, particularly N−H⋅⋅⋅N and C−H⋅⋅⋅π interactions, which influence the overall crystal packing. The target bis‐hydrazone exhibited significant antimicrobial activity against multi‐drug‐resistant bacterial strains, with the largest activity against S.typhimurium with an inhibition zone of 17.1±0.6 mm and MIC 31.25 μg/mL. The compound also demonstrated significant cytotoxic effects on cancer cells, with a higher IC50 ratio of 134.43 μg/mL against the normal cell line Wi38 and the lowest IC50 value of 45.88 μg/mL against the cancer cell line Caco2. Molecular docking was carried out with estrogen receptor alpha (ERα) and sodium‐glucose transporter SGLT1, which are relevant to Mcf7 and Caco2 cancer cell lines, respectively. Docking suggests the presence of specific amino acids that may influence the binding affinity between the ligand and receptor active sites through residue overlaps in chains A for SGLT1 and B for Erα, offering the ligand as a promising anticancer consistent with the IC50 outcomes.

Enhancement of apoptosis in Caco-2, Hep-G2, and HT29 cancer cell lines following exposure to Toxoplasma gondii peptides.

Cancer or neoplasm is a cosmopolitan catastrophe that results in more than 20 million new cases and 10 million deaths every year. Some factors lead to carcinogenesis like infectious diseases. Parasites like Toxoplasma gondii, by its components, could modulate the cancer system by inducing apoptosis. The objective of this investigation is to assess the potential of peptides derived from T. gondii in combating cancer by examining their effects on Caco-2, Hep-G2, and HT29 cell lines. Candidate peptide by its similarity to anticancer compounds was predicted through the computer-based analysis/platform. The impact of the peptide on cell viability, cell proliferation, and gene expression was evaluated through the utilization of MTT assay, flow cytometry, and real-time polymerase chain reaction (PCR) methodologies. The cell viability rate exhibited a significant decrease (p < 0.001) across all cell lines when exposed to a concentration of ≤160 μg. Within the 48-hour timeframe, the half maximal inhibitory concentration (IC50) for HT29 and Hep-G2 cell lines was determined to be 107.2 and 140.6 μg/mL, respectively. Notably, a marked decrease in the expression levels of Bcl2 and APAF1 genes was observed in both the Hep-G2 and HT29 cell lines. These findings indicate that the T. gondii peptide affected cancer cell mortality and led to changes in the expression of genes associated with apoptosis.

Preparation of new compounds with anticancer properties via the reaction of Kryptofix 5 with two organic acceptors

The new designed complexes via CT interaction between kryptofix 5 and two organic acceptors to introduce basic data that can be used to assessment of crown ether and tested the final complexes against cancer cells. This target was achieved by synthesis of a new CT- complexes of kryptofix 5 as a type of mixed nitrogen–oxygen crown ethers with π-acceptors as (2 hydroxy-3.5-dinitrobenzoic acid and 2,3-dichloro-5,6-dicyano-1,4-benzoquinone). These obtained complexes were structurally characterized in both solid and liquid state via various chemical methods as elemental analysis, FT-IR, NMR, and UV–Visible spectroscopy, thermal degradation and physical properties were calculated. Finally, the CT complex was also tested for its anticancer activity against two human cancer cell lines; colon cancer CaCo2 and liver cancer cell line (hepG2). KEY WORDS: Anticancer, Kryptofix 5, HDNS, DDQ, Spectroscopy Bull. Chem. Soc. Ethiop. 2024, 38(6), 1639-1651. DOI: https://dx.doi.org/10.4314/bcse.v38i6.11

Phenolic changes in propolis during in vitro digestion and cytotoxic effects on human cancer cell lines

PurposeThis study aimed to evaluate the compositional changes and bioaccessibility of phenolics and antioxidants in propolis during in vitro digestion as well as the cytotoxic effects of digested propolis on various cancer cell lines.Design/methodology/approachSix propolis samples were obtained and subjected to in vitro oral, gastric and intestinal digestion. Both digested and undigested samples were analyzed for their total phenolic, flavonoid and antioxidant activities. Additionally, changes in phenolic composition in the in vitro digestion system were revealed by the HPLC-DAD system. The cytotoxic effects of the digested samples were assessed on lung (A549, H1299), skin (A431), liver (Hep-G2) and colon (Caco-2) cancer cells as well as on fibroblast (Bj) cells.FindingsThe mean bioaccessibility values of phenolic and flavonoid compounds were found to be less than 35 and 24%, respectively, while the TEAC and CUPRAC antioxidant results ranged between 225.08–649.04 and 398.68–1552.28 µmol TE/g, respectively. The release of p-coumaric, ferulic, 3,4-dimethoxycinnamic acids, naringenin, pinocembrin and chrysin increased progressively from the oral to the intestinal stage. The cytotoxic effects of samples on cell lines were ranked, based on IC50 results, as A431 > Hep-G2 > Caco-2 > A549 > H1299 > Bj.Originality/valuePropolis has been recognized for centuries as a natural remedy, and numerous studies have explored its bioactive components. However, no studies have previously examined the changes in the phenolic compositions of propolis samples during digestion or their cytotoxic effects on cancer cells. Therefore, this study provides novel insights and an approach to the existing literature on this topic.Graphical abstract

Nuciferol C, a new sesquineolignan dimer from Cocos nucifera L.: bioactivity and theoretical investigation.

Nuciferol C (NC), an undescribed dimer of nuciferol B (NB), was isolated from the endocarp of Cocos nucifera L. The planar structure of NC was determined using 1D- and 2D-NMR spectroscopy as well as high resolution MS spectrometry. The absolute configuration was concluded based on analysis of NOESY spectra. NC showed cytotoxic activity against colon cancer cells (CaCo-2) with an IC50 value of 76 μM, and significantly decreased the expression of human epidermal growth factor receptor (EGFR) and tumor necrosis factor alpha (TNF-α) in CaCo-2 as compared with untreated cells by 39% and 33%, respectively (p < 0.05). In addition, NC exhibited anti-herpes simplex virus (HSV-I) activity with an IC50 value of 23 μM. In silico study of NC was implemented at three levels: density functional theory (DFT) was used to study its electronic properties, molecular mechanics was used to estimate the docking results, and finally, molecular dynamic simulation was used to study the behavior and stability of NC inside the active site of the target protein of HSV-1.

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.

Antimicrobial and cytotoxic activities of flavonoid and phenolics extracted from Sepia pharaonis ink (Mollusca: Cephalopoda)

BackgroundSeveral studies have been reported previously on the bioactivities of different extracts of marine molluscs. Therefore, we decided to evaluate the cytotoxic and antimicrobial activities of S. pharaonis ink as a highly populated species in the Red Sea. We extracted the flavonoids from the ink and analyzed their composition. Then we evaluated systematically the cytotoxic and antimicrobial properties of this extract. A pharmacokinetic study was also conducted using SwissADME to assess the potential of the identified flavonoids and phenolic compounds from the ink extract to be orally active drug candidates.ResultsCytotoxic activity was evaluated against 5 cell lines (MCF7, Hep G2, A549, and Caco2) at different concentrations (0.4 µg/mL, 1.6 µg/mL, 6.3 µg/mL, 25 µg/mL, 100 µg/mL). The viability of examined cells was reduced by the extract in a concentration-dependent manner. The highest cytotoxic effect of the extract was recorded against A549 and Hep G2 cancer cell lines cells with IC50 = 2.873 and 7.1 µg/mL respectively. The mechanistic analysis by flow cytometry of this extract on cell cycle progression and apoptosis induction indicated that the extract arrests the cell cycle at the S phase in Hep G2 and MCF7, while in A549 cell arrest was recorded at G1 phase. However, it causes G1 and S phase arrest in Caco2 cancer cell line. Our data showed that the extract has significant antimicrobial activity against all tested human microbial pathogens. However, the best inhibitory effect was observed against Candida albicans ATCC 10,221 with a minimum inhibitory concentration (MIC) of 1.95 µg/mL. Pharmacokinetic analysis using SwissADME showed that most flavonoids and phenolics compounds have high drug similarity as they satisfy Lipinski’s criteria and have WLOGP values below 5.88 and TPSA below 131.6 Å2.ConclusionS. pharaonis ink ethanolic extract showed a promising cytotoxic potency against various cell lines and a remarkable antimicrobial action against different pathogenic microbial strains. S. pharaonis ink is a novel source of important flavonoids that could be used in the future in different applications as a naturally safe and feasible alternative of synthetic drugs.

Tripeptide linked dispiro cyclotriphosphazene conjugates: Synthesis, molecular docking analysis of compounds binding within cancer cell line receptors and in vitro cytotoxic and genotoxic activities

The novel dioxybiphenyl bridged-cyclotriphosphazenes (DPP) bearing tripeptide were synthesized and investigated for their molecular docking analysis, visualizing their binding profiles within various cancer cell line receptors and in vitro cytotoxic and genotoxic properties. The dipeptide compound (Tyr-Phe) was treated with various amino acids to obtain the tripeptide compounds (Tyr-Phe-Gly, Tyr-Phe-Ala, Tyr-Phe-Val, Tyr-Phe-Phe, and Tyr-Phe-Leu). These synthesized tripeptides were subsequently treated with DPP to obtain novel phosphazene compounds bearing tripeptide structures. As a result, the synthesis of target molecules with phosphazene compound in the center and biphenyl and tripeptide groups in the side arms was obtained for the first time in this study. Examining the cytotoxic studies in vitro of our newly synthesized compounds demonstrated the anticancer properties against four selected human cancer cell lines, including breast (MCF-7), ovarian (A2780), prostate (PC-3), and colon (Caco-2) cancer cells. The Comet Assay analysis determined that the cell death mechanism of most of the compounds with cytotoxic activity stemmed from the DNA damage mechanism. Among the compounds, the DPP-Tyr-Phe-Phe compound seems to have the best anticancer activity against the subjected cell lines (Except for A2780) with IC50 values equal to 20.18, 72.14, 12.21, and 5.17 μM against breast, ovarian, prostate, and colon cancer cell lines, respectively. For this reason, the molecular docking analysis was conducted for the DTPP compound to visualize its binding geometry and profile within the target enzyme’s binding site associated with the specific cancer cell line. The analysis revealed that the DTPP derivative exhibited an optimal binding conformation and characteristics within the target enzyme’s binding site, aligning well with the experimental data. Based on the data, these compounds are believed to be strong candidate molecules for both pharmaceutical and clinical applications.

Transforming microbial pigment into therapeutic revelation: extraction and characterization of pyocyanin from Pseudomonas aeruginosa and its therapeutic potential as an antibacterial and anticancer agent

BackgroundThe objectives of the current study were to extract pyocyanin from Pseudomonas aeruginosa clinical isolates, characterize its chemical nature, and assess its biological activity against different bacteria and cancer cells. Due to its diverse bioactive properties, pyocyanin, being one of the virulence factors of P. aeruginosa, holds a promising, safe, and available therapeutic potential.Methods30 clinical P. aeruginosa isolates were collected from different sources of infections and identified by routine methods, the VITEK 2 compact system, and 16 S rRNA. The phenazine-modifying genes (phzM, phzS) were identified using polymerase chain reaction (PCR). Pyocyanin chemical characterization included UV-Vis spectrophotometry, Fourier Transform Infra-Red spectroscopy (FTIR), Gas Chromatography-Mass Spectrometry (GC-MS), and Liquid Chromatography-Mass Spectrometry (LC-MS). The biological activity of pyocyanin was explored by determining the MIC values against different clinical bacterial strains and assessing its anticancer activity against A549, MDA-MB-231, and Caco-2 cancer cell lines using cytotoxicity, wound healing and colony forming assays.ResultsAll identified isolates harboured at least one of the phzM or phzS genes. The co-presence of both genes was demonstrated in 13 isolates. The UV-VIS absorbance peaks were maxima at 215, 265, 385, and 520 nm. FTIR could identify the characteristic pyocyanin functional groups, whereas both GC-MS and LC-MS elucidated the chemical formula C11H18N2O2, with a molecular weight 210. The quadri-technical analytical approaches confirmed the chemical nature of the extracted pyocyanin. The extract showed broad-spectrum antibacterial activity, with the greatest activity against Bacillus, Staphylococcus, and Streptococcus species (MICs 31.25–125 µg/mL), followed by E. coli isolates (MICs 250–1000 µg/mL). Regarding the anticancer activity, the pyocyanin extract showed IC50 values against A549, MDA-MB-231, and Caco-2 cancer cell lines of 130, 105, and 187.9 µg/mL, respectively. Furthermore, pyocyanin has markedly suppressed colony formation and migratory abilities in these cells.ConclusionsThe extracted pyocyanin has demonstrated to be a potentially effective candidate against various bacterial infections and cancers. Hence, the current findings could contribute to producing this natural compound easily through an affordable method. Nonetheless, future studies are required to investigate pyocyanin’s effects in vivo and analyse the results of combining it with other traditional antibiotics or anticancer drugs.