Studies on Anticancer Effect of Methanolic Leaf Extract of Justicia gendarussa on Lung Cancer Cell Line.

Objective: The aim of study's goal was to look into the anticancer efficacy of a methanolic extract of Justicia gendarussa against a lung cancer cell line. Materials and Methods: Cell viability assays and cell and nuclear morphology examinations were used to evaluate the anticancer efficacy against methanolic extract of Justicia gendarussa on lung cancer cell lines. The IC50 doses were calculated using different concentrations of Justicia gendarussa extract (0, 10, 20, 40, 60, and 80 µg/mL). Results: The results of MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide) assay revealed that the percentage of viability in treated cells was significantly lower as compared with untreated control groups, which represented as 100%, and an inhibitory concentration of 40 µg/mL was observed. Under a phase-contrast microscope, morphological changes revealed cell shrinkage and cytoplasmic membrane blebbing. The apoptotic nuclei (intensely colored, broken nuclei, and compacted chromatin) were examined under a fluorescence microscope. Conclusions: The outcome of the research work on Justicia gendarussa was investigated for anticancer properties. The results revealed the proapoptotic and cytotoxic effects of Justicia gendarussa extract on lung cancer cell lines. From the above results and findings, it could be concluded that the Justicia gendarussa methanolic leaf extract exhibited potent anticancer activity against a lung cancer cell line. Further study needs to be conducted to investigate the active chemicals in the extract as well as the molecular mechanisms underlying its anticancer benefits.

Evaluation of In vitro Anti-Cancer Activity of Methanolic Leaf Extract of Phoenix pusilla on Colon Cancer Cell Line.

Background: Cancer rates continue to climb, owing largely to the world population's aging and growth, as well as economically developing countries, a surge in cancer-causing behavior, particularly smoking. The third or fourth most prevalent type of cancer is colon cancer. Cancer of the large intestine (colon) is one of the primary causes of death from cancer. Colorectal cancer prevention is mostly based on adenomatous disease screening approaches. The cytotoxic and pharmacological properties of Phoenix pusilla are widely documented. As a result, there is little recorded evidence of its cytotoxic activity against colon cancer cells. Therefore, we planned to study the efficacy of a methanolic leaf extract of Phoenix pusilla against in vitro colon cancer cells. Aim: To evaluate the anti-cancer effects of the methanolic leaf extract of Phoenix pusilla on colon cancer cell lines. Materials and Methods: In vitro screening and anti-cancer effects of the methanolic effect of Phoenix pusilla on colon cancer cell lines were assessed by cell viability assays and cell and nuclear morphological studies. For the in vitro cell culture study, different concentrations of Phoenix pusilla leaf extract (0, 25, 50, 75, 100, 150 µg/ml) were used, and IC50 doses were calculated. Results: The results of the MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) assay revealed that the fraction of viability cells significantly decreased in treated cells when compared to untreated control groups, was expressed as 100%, and an inhibitory concentration of µg/ml was identified. A phase-contrast microscope was used to observe cell shrinkage and cytoplasmic membrane blebbing. A fluorescent microscope was used to examine the apoptotic nuclei (internally dyed nuclei, shattered nuclei, and condensed chromatin). Conclusion: In conclusion, the present study results showed that the leaf extracts of Phoenix pusilla had a strong cytotoxic effect and induced significant apoptosis in the colon cancer cell lines at a concentration of 75 µg/ml in the 24 h incubation period. More research is needed to investigate the extract's active components as well as the molecular mechanisms underlying its anti-cancer properties.

Human Colon Cancer HT29 Cell Line Treatment with High-Dose LAscorbic Acid Results to Reduced Angiogenic Proteins Expression and Elevated Pro-apoptotic Proteins Expression.

BACKGROUND: Some studies have shown anticarcinogenic effects of high dose L-Ascorbic Acid. However, there are controversies around the therapeutic administration of Ascorbic acid as an anticancer medicine. OBJECTIVE: We conducted a case-control study to investigate the role of pharmacologic concentration of Ascorbic acid on viability and angiogenesis of the human colon cancer (HT29) cell line. METHODS: The HT29 cells were cultured in DMEM-HG and treated with 10 mM ascorbic acid for 3h. The culture medium was exchanged, and after incubation at 37 ºC for 24 h, the cells were collected and utilized to evaluate viability, ROS production, gene expression and protein expression levels. The control group consisted of untreated HT29 cells. The viability of the cells was determined using the MTT method. Moreover, Nitro Blue Tetrazolium (NBT) was used to detect the ROS production capacity. The mRNA transcript's level and protein expression were evaluated by Real-time PCR and Western blotting, respectively. RESULTS: The ascorbic acid-treated group showed a significant increase in ROS production and an obvious reduction in viability compared to the control group. The treated group showed significantly increased levels of both early apoptotic markers (Bax, Cyt C, Caspase3, and Caspase 9) and late apoptotic markers (Caspase 8). Bcl2 expression showed significantly decreased levels relative to the control group. Ascorbic acid therapy substantially reduced the expression of bFGF, bFGFR, PDGF, PDGFR and PLC- γ compared to the control group. CONCLUSION: The results confirm that high-dose L-ascorbic acid reduces HT29 cell line viability in vitro.

Microbial Azurin Immobilized on Nano-Chitosan as Anticancer and Antibacterial Agent Against Gastrointestinal Cancers and Related Bacteria.

OBJECTIVES: To discover new natural effective anticancer agents and new antibacterial agents against antibiotic-resistant bacteria which are the most serious public health concern. Another important concern is drug delivery which is the transport of pharmaceutical compounds to have a therapeutic effect in organisms having a disease. Azurin is a promising anticancer agent produced from Pseudomonas aeruginosa. This study tried to test the effectiveness of the immobilization of azurin on nano-chitosan to enhance its anticancer and antibacterial activity against gastrointestinal cancer and its related bacteria. METHODS: We purified azurin protein from Pseudomonas aeruginosa and then immobilized it on nano-chitosan. The anticancer activity of the free and nano-azurin is tested against a gastric cancer cell line (CLS-145), pancreatic cancer cell line (AsPC-1), colon cancer cell line (HCT116), esophagus cancer cell line (KYSE-410), and liver cancer cell line (HepG2). The antibacterial activity of both free and immobilized azurin also is tested against bacterial species related to the gastrointestinal cancer biopsies: Helicobacter pylori, Bacteroides fragilis, Salmonella enterica, Fusobacterium nucleatum, and Porphyromonas gingivalis. RESULTS: Both free and nano-azurin showed high anticancer and antibacterial activity. Immobilization significantly increased the anticancer and antibacterial activity of the azurin CONCLUSION: Nano-azurin can be used as an effective anticancer and antibacterial agent against gastrointestinal cancer and bacterial species related to these cancers.

Peroxiporins Are Induced upon Oxidative Stress Insult and Are Associated with Oxidative Stress Resistance in Colon Cancer Cell Lines.

Oxidative stress can induce genetic instability and change cellular processes, resulting in colorectal cancer. Additionally, adaptation of oxidative defense causes therapy resistance, a major obstacle in successful cancer treatment. Peroxiporins are aquaporin membrane channels that facilitate H2O2 membrane permeation, crucial for regulating cell proliferation and antioxidative defense. Here, we investigated four colon cancer cell lines (Caco-2, HT-29, SW620, and HCT 116) for their sensitivity to H2O2, cellular antioxidative status, and ROS intracellular accumulation after H2O2 treatment. The expression of peroxiporins AQP1, AQP3, and AQP5 and levels of NRF2, the antioxidant transcription factor, and PPARγ, a transcription factor that regulates lipid metabolism, were evaluated before and after oxidative insult. Of the four tested cell lines, HT-29 was the most resistant and showed the highest expression of all tested peroxiporins and the lowest levels of intracellular ROS, without differences in GSH levels, catalase activity, nor NF2 and PPARγ levels. Caco-2 shows high expression of AQP3 and similar resistance as HT-29. These results imply that oxidative stress resistance can be obtained by several mechanisms other than the antioxidant defense system. Regulation of intracellular ROS through modulation of peroxiporin expression may represent an additional strategy to target the therapy resistance of cancer cells.

Extraction of phenolic compounds from the shells of pecan nuts with cytotoxic activity through apoptosis against the colon cancer cell line HT-29.

The water extraction of phenolic compounds from two varieties ("Mahan" and "Marameck") of pecan nutshells (Carya illinoinensis) without and with sonication, varying the solvent/solid ratio (S), the pH, and the refluxing time (t), was studied. Additionally, the in vitro cytotoxicity and the determination of the cell death mechanism of the extracts against the colon cancer cell line HT-29 were investigated. The content of total phenolic compounds (TPC) of "Marameck" nutshells resulted higher than for the "Mahan" variety, and the pH increase resulted in higher TPC contents for both cultivars. The optimized conditions for TPC extraction without and with sonication resulted: S = 33 ml/g, pH = 12, and t = 9.6 min, and yielded ≈ 70 and 90 mg/g of TPC for "Mahan" and "Marameck" nutshells, respectively. The optimized extracts of pecan nutshells without sonication from both cultivars presented similar cytotoxicity against HT-29 colon cancer cells (IC50  ≈ 50 µg/ml), higher than for sonicated extracts (IC50 ≈ 88 and 138 µg/ml for "Mahan" and "Marameck," respectively). Cell death through apoptosis was the main mechanism of cell death induced by the nutshell extracts. PRACTICAL APPLICATION: The extraction of phenolic compounds (TPC) from the residues of two varieties of pecan nutshells ("Mahan" and "Marameck") was studied. An optimal combination of variables within the pH range that minimizes the solvent-to-solid ratio (S) and the time of refluxing (t), saving at the same time, water and energy, was set up. The phenolic compound extracts obtained from the residues of the pecan nuts exhibit cytotoxic effects against colon cancer cells and could be of interest as an alternative treatment of different types of cancer. Additionally, these extracts may be of importance to the food industry as they can be used as antioxidant agents in food formulation. Also, the high levels of anthocyanidins obtained from the pecan nut extracts after proanthocyanidins' strong acid hydrolysis can be purified and employed as natural red dyes.

Anticancer potential of ascorbic acid and inorganic selenium on human breast cancer cell line MCF-7 and colon carcinoma HCT-116.

BACKGROUND AND OBJECTIVES: Till now, cancer is a major health problem and one of the main causes of mortality worldwide. Ascorbic acid and selenium are the two most popular dietary supplements used to prevent cancer proliferative, therefore, the work aims to study the antitumor effect of ascorbic acid and selenium on HCT116 and MCF7 cell lines. MATERIALS AND METHODS: In the present study, the cytotoxic effect of different concentrations of ascorbic acid and selenium on human breast cancer cell line (MCF7 cells) and human colon carcinoma (HCT116) was studied. RESULTS: Viability % of HCT116 cell line and MCF7 cell line decreased with increasing ascorbic acid concentrations (1-4 mM). The 50% inhibitory concentration (IC50) of five dilutions of each concentration of ascorbic acid was evaluated in the current study. IC50 was 0.18, 0.17, 0.16, and 0.16 mM for HCT116 cell line and was 0.86, 1.34, 1.74, and 0.47 mM for MCF7 cell line at 1, 2, 3, and 4 mM, respectively. Cell viability decreased depending on the selenium concentrations ranging from 20 to 100 mM. Selenium effect showed less cytotoxicity on MCF7 compared to HCT116 cells at all tested concentrations where the cell viability at 20, 40, 60, 80, and 100 mM selenium was 33.74, 29.48, 26.08, 54.53, and 20.89 for HCT116 cell and was 79.53, 76.01, 59.42, 54.53, and 51.98 for MCF7 cell, respectively. Ascorbic acid induced apoptosis by promoting the release of lactate dehydrogenase (LDH) in HCT116 and MCF7 cells, but reduced release of LDH was observed in selenium treatment but increased when it added to ascorbic acid because of a possible synergistic action that may produce an enhanced anticarcinogenic effect. CONCLUSION: The present study documented that a combination of ascorbic acid and selenium produces an additive chemopreventive effect on carcinogenesis.

Biological Effect of Different Spinach Extracts in Comparison with the Individual Components of the Phytocomplex.

The Mediterranean-style diet is rich in fruit and vegetables and has a great impact on the prevention of major chronic diseases, such as cardiovascular diseases and cancer. In this work we investigated the ability of spinach extracts obtained by different extraction methods and of the single main components of the phytocomplex, alone or mixed, to modulate proliferation, antioxidant defense, and genotoxicity of HT29 human colorectal cells. Spinach extracts show dose-dependent activity, increasing the level of intracellular endogenous reactive oxygen species (ROS) when tested at higher doses. In the presence of oxidative stress, the activity is related to the oxidizing agent involved (H2O2 or menadione) and by the extraction method. The single components of the phytocomplex, alone or mixed, do not alter the intracellular endogenous level of ROS but again, in the presence of an oxidative insult, the modulation of antioxidant defense depends on the oxidizing agent used. The application of the phytocomplex extracts seem to be more effective than the application of the single phytocomplex components.

Anti-Tumor Mechanisms of Novel 3-(4-Substituted Benzyl)-5-Isopropil-5- Phenylhydantoin Derivatives in Human Colon Cancer Cell Line.

BACKGROUND: Hydantoin and its newly synthesized derivatives have recently become a focus of interest due to their numerous biological activities and newly emerging beneficial effects in different pathological conditions, including cancer. OBJECTIVE: The aim of this study was to evaluate the possible anti-tumor mechanisms of a series of newly synthesized 3-(4-substituted benzyl)-5-isopropyl-5-phenylhydantoin derivatives in different aspects of cell physiology of human colon cancer cell line, HCT-116. METHODS: The increasing concentrations of derivatives (0.01µM up to 100µM) were applied to cells during 24h, 48h, and 72h after which the evaluation of proliferation, apoptosis, oxidative/anti-oxidative status, nitrite production, and migration/invasion potential of treated cells was performed. RESULTS: All tested compounds expressed the dose- and time-dependent anti-proliferative and pro-apoptotic activities against HCT-116 cells. The investigated derivatives induced a decrease in levels of oxidative stress parameters and an increase in levels of nitrite production by treated cells suggesting their significant antioxidative effects. The cell migration index and expression level of tumor invasion-promoting metalloproteinase- 9 (MMP-9) gene were significantly decreased after treatment with the tested hydantoin derivatives implicating their inhibitory role in colon cancer cell motility and invasion processes. The mRNA level of cyclooxygenase-2 (COX-2) gene as a pro-inflammatory gene related to colorectal carcinogenesis was reduced compared to values in the non-treated control cells indicating the significant anti-inflammatory/anti-tumor effects of these compounds. CONCLUSION: The obtained results show the significant anti-tumor potential of tested derivatives, especially 3- benzyl-5-isopropyl-5-phenylhydantoin and 3-(4-chlorobenzyl)-5-isopropyl-5-phenylhydantoin, suggesting their potential usage in the development of more effective chemotherapies.

Metal complexes driven from Schiff bases and semicarbazones for biomedical and allied applications: a review.

Schiff bases are versatile organic compounds which are widely used and synthesized by condensation reaction of different amino compound with aldehydes or ketones known as imine. Schiff base ligands are considered as privileged ligands as they are simply synthesized by condensation. They show broad range of application in medicine, pharmacy, coordination chemistry, biological activities, industries, food packages, dyes, and polymer and also used as an O2 detector. Semicarbazone is an imine derivative which is derived from condensation of semicarbazide and suitable aldehyde and ketone. Imine ligand-containing transition metal complexes such as copper, zinc, and cadmium have shown to be excellent precursors for synthesis of metal or metal chalcogenide nanoparticles. In recent years, the researchers have attracted enormous attention toward Schiff bases, semicarbazones, thiosemicarbazones, and their metal complexes owing to numerous applications in pharmacology such as antiviral, antifungal, antimicrobial, antimalarial, antituberculosis, anticancer, anti-HIV, catalytic application in oxidation of organic compounds, and nanotechnology. In this review, we summarize the synthesis, structural, biological, and catalytic application of Schiff bases as well as their metal complexes.

Bioassay-guided isolation of cytotoxic compounds from Chrysophthalmum montanum (DC.) Boiss.

Bioassay-guided isolation of the 80% methanol extract of the aerial parts of Chrysophthalmum montanum (DC.) Boiss. (Asteraceae) led to the isolation of four known guaianolide-type sesquiterpene lactones, 6α-acetoxy-4α-hydroxy-1ßH-guaia-9.11(13)-dien-12.8α-olide (1), 6α-acetoxy-4α-hydroxy-9ß.10ß-epoxy-1ßH-guaia-11(13)-en-12.8α-olide (2), 4α,6α-dihydroxy-1ß,5α,7αH-guaia-9(10),11(13)-dien-12,8α-olide (3), and (4α,5α,8ß,10ß)-4,10-dihydroxy-1,11(13)-guaidien-12,8-olide (4), along a steroidal glycoside mixture (5a and 5b). The structures of the compounds were identified on the basis of spectroscopic data. Among them, 2, 4 and a steroidal glycoside mixture were obtained from C. montanum for the first time. All isolates were also first time assayed for in vitro cytotoxicities against four human cancer cell lines, i.e. breast (MCF-7, MDA-MB 231), colon (HT-29), and lung (PC3). Among the isolates, 1-3 showed significant inhibitory effect on the proliferation of cancer cells with viability ranging from 6.86 to 26.51%, while steroidal glycoside mixture showed no cytotoxicity, except against HT-29 (viability 61.99%). Compound 4 exhibited strong and selective cell growth inhibition against HT-29 with viability 20.99% and was identified as a promising compound with high selectivity between cancer cells and normal human lung cells (BEAS-2B), especially against HT-29 (IC50 = 12.2 µg/mL) compared to that of cisplatin. These results suggested that 4 is worthy of further study to determine its cytotoxicity mechanisms.

Study on Synthesis and Anti-tumor Activity of 6 Kinds of Tectorigenin Mannich Base Derivatives / 中国药房

OBJECTIVE： To conduct structural modification of tectorigenin to search for new compounds with anti-tumor activity. METHODS： Tectorigenin was used as a lead compound， and then added into amine reagents as ethanolamine， methylamine， ethylamine， dimethylamine， diethylamine， n-propylamine and formaldehyde solution. Tectorigenin Mannich base derivatives were synthesized by mannich reaction with as the lead compound. The structures of the derivatives were identified according to IR， UV， MS and NMR data. Solubility of tectorigenin and its derivatives were investigated by solubility test method. MTT assay was used to investigate the inhibitory effects of tectorigenin and its derivatives on the proliferation of human colon cancer cell line HCT116， human lung cancer cell line A549 and human hepatoma cell line HepG2， and half inhibitory concentration （IC50） was calculated. The inhibition rate of tectorigenin and its derivatives （100 mg/kg） on H22 hepatoma-bearing mice in vivo was studied. RESULTS： Totally of 6 kinds of tectorigenin mannich base derivatives were synthesized， such as 8-（N-hydroxyethyl）-methyleneamino-5，7，4′-trihydroxy-6-methoxyisoflavone， 8-（N-methyl）-methyleneamino-5，7，4′-trihydroxy-6- methoxyisoflavone， 8-（N， N-diethyl）-methyleneamino-5，7，4′-trihydroxy-6-methoxyisoflavone， 8-（N， N-dimethyl）-methyleneamino- 5，7，4′-trihydroxy-6-methoxyisoflavone， 8-（N-ethyl）-methyleneamino-5，7，4′-trihydroxy-6-methoxyisoflavone， 8-（N-propyl）- methyleneamino-5，7，4′-trihydroxy-6-methoxyisoflavone （compounds 1-6 in turn）. Compared with tectorigenin， the water solubility of six derivatives was significantly improved， and the solubility was 5-20 times higher than that of tectorigenin. IC50 of compounds 1， 3 and 5 to HCT116 cells were （34.82±3.27）， （16.21±4.13）， （33.12±3.25） μmol/L， which were stronger than that of tectorigenin [（45.23±5.74） μmol/L]； IC50 of compounds 1， 3 and 5 to A549 cells were （37.05±5.74）， （26.88±4.52）， （30.13±6.23） μmol/L， which were stronger than that of tectorigenin [（53.24±6.34） μmol/L]； IC50 of compounds 1， 3 and 5 to HepG2 cells were （23.74±1.45）， （18.96±2.34）， （30.95±2.87） μmol/L， which were stronger than that of tectorigenin [（48.98±2.58） μmol/L]. Compounds 1， 3 and 5 showed higher inhibition rates （55.51％， 57.20％ and 49.15％） than tectorigenin （33.05％） on H22 hepatoma-bearing mice， respectively. The other three compounds had no obvious advantage over tectorigenin in anti-tumor activity. CONCLUSIONS： In this study， compounds 1， 3 and 5 of six tectorigenin mannich base derivatives synthesized in this study have stronger antitumor activity than tectorigenin.

Thermal Co-reduction engineered silver nanoparticles induce oxidative cell damage in human colon cancer cells through inhibition of reduced glutathione and induction of mitochondria-involved apoptosis.

Silver nanoparticles (AgNPs) are being commercialized in a number of consumer products including food and cosmetics where there is a direct exposure of AgNPs to human body. An extensive toxicological evaluation is necessary to understand the mechanism for its safe use, since the toxicity effect varies greatly with the synthesis protocol followed. In this study, we report the detailed toxicological analysis of AgNPs fabricated by thermal co-reduction approach. Our study was analysed in human colon cancer cell line (HCT 116) and the IC50 was calculated as 28.11 µg/ml. It was also observed that AgNP induces oxidative stress on HCT116 by increased levels of lipid peroxidation and reduced levels of glutathione. Mitochondrial membrane depolarization was also analysed and Western blot analysis confirms the increased level of Bcl and Caspase-3 which indicates the mitochondrial -mediated apoptosis. Additionally, flow cytometric analysis suggests cell cycle arrest in G2/M phase. Thus, our study can be a basis for further research to design safe AgNPs in various consumer products. Additionally, similar research can be conducted for different size and shape of AgNP or nano-silver can be engineered using different approaches.

Bioaccessibility during In Vitro Digestion and Antiproliferative Effect of Bioactive Compounds from Andean Berry ( Vaccinium meridionale Swartz) Juice.

Berry consumption is associated with colorectal-cancer chemoprevention, but digestive conditions can affect this property. The bioaccessibility and apparent permeability coefficients of bioactive compounds from Andean Berry Juice (ABJ) after in vitro gastrointestinal digestion and colonic fermentation were analyzed. The antiproliferative effect of the fermented nondigestible fraction was evaluated against SW480 colon-adenocarcinoma cells. Gallic acid displayed the highest bioaccessibility in the mouth, stomach, small intestine, and colon. However, chlorogenic acid exhibited the highest apparent permeability coefficients (up to 1.98 × 10-4 cm/s). The colonic-fermentation fraction showed an increase of ≥50% antiproliferative activity against SW480 cells (19.32%, v/v), equivalent to those of gallic acid (13.04 µg/g), chlorogenic acid (7.07 µg/g), caffeic acid (0.40 µg/g), ellagic acid (7.32 µg/g), rutin (6.50 µg/g), raffinose (0.14 mg/g), stachyose (0.70 mg/g), and xylose (9.41 mg/g). Bioactive compounds from ABJ are bioaccessible through the gastrointestinal tract and colon fermentation, resulting in antiproliferative activity.

Inhibitory effects of aesculetin on the proliferation of colon cancer cells by the Wnt/ß-catenin signaling pathway.

The inhibitory effect of aesculetin on the growth of colon cancer cell line SW480 through the Wnt/ß-catenin signaling pathway was studied. The appropriate concentration of aesculetin was selected by cell counting kit-8 (CCK-8) assay, and the effect of aesculetin on the proliferation of SW480 cells was investigated by bromodeoxyuridine (BrdU) assay. The expression level of the messenger ribonucleic acid (mRNA) in ß-catenin and Wnt signaling pathway target genes, c-Myc and cyclin D1, was detected by reverse transcription-polymerase chain reaction (RT-PCR). The expression levels of ß-catenin, c-Myc and cyclin D1 proteins were detected by western blotting. CCK-8 detection results showed that compared with the control group, aesculetin effectively inhibited the proliferation of SW480 cells. BrdU detection results indicated that the number of BrdU positive cells in all the groups treated with drugs was significantly decreased. The of RT-PCR results suggested that aesculetin reduced the expression level of ß-catenin mRNA and inhibited the expression of mRNA in the Wnt signaling pathway target genes, c-Myc and cyclin D1. Western blotting detection results revealed that aesculetin downregulated the expression level of ß-catenin, c-Myc and cyclin D1 proteins. Aesculetin can inhibit tumor growth by suppressing the Wnt signaling pathway. This study provides a new idea and direction for the antitumor mechanism of aesculetin.

Green synthesis of silver nanoparticles using Anthemis atropatana extract: characterization and in vitro biological activities.

This study aim to synthesize silver nanoparticles (AgNPs) using Anthemis atropatana extract and to evaluate their chemical characteristics and antimicrobial and cytotoxic effects. The biosynthesis of AgNPs is verified using UV-visible spectrum which showing maximum absorption in 430 nm wavelength. Transmission electron microscope (TEM) and scanning electron microscope (SEM) results revealed that AgNPs has a spherical shape with an average size of 38.89 nanometres. The crystalline structure of green synthesized AgNPs in optimal conditions was confirmed by XRD analysis. The pattern of XRD peaks related to face-centred cubic (fcc) (111), (200), (220), (311) and (222) observed. Also, FTIR results verified the AgNPs synthesis using plant extract. In biological tests, the MTT results indicate the dose dependence of cytotoxic effects of AgNPs on colon cancer cell lines (HT29). The AgNPs had maximum cytotoxicity on HT29 cancer cell line at 100 µg/ml concentration, which were statistically significant comparing control cells (p < .001). Moreover, real time PCR and flow cytometry results confirmed the apoptotic effects of AgNPs. According to the results, it seems that the green synthesis of AgNPs is an eco-friendly and cost effective approach. This research provides insight into the development of new anticancer and antibacterial agents.

Oxidative stress-induced mitochondrial dysfunction in a normal colon epithelial cell line.

AIM: To determine how a normal human colon cell line reacts to microbial challenge as a way to study oxidative stress-induced responses associated with inflammatory bowel disease. METHODS: Normal human colon epithelial cells (ATCC® CRL.1790™) were stimulated with either heat killed E. coli or heat killed murine cecal contents (HKC) and examined for several relevant biomarkers associated with inflammation and oxidative stress including cytokine production, mitochondrial autophagy and oxidant status. TNFα, IL-1ß and IL-8 protein concentrations were measured within the supernatants. Fluorescent microscopy was performed to quantify the production of reactive oxygen species (ROS) using an oxidation responsive fluorogenic probe. Mitochondrial morphology and mitochondrial membrane potential was assessed by dual staining using COXIV antibody and a dye concentrating in active mitochondria. Mitochondrial ROS scavenger was used to determine the source of ROS in stimulated cells. Autophagy was detected by staining for the presence of autophagic vesicles. Positive controls for autophagy and ROS/RNS experiments were treated with rapamycin and chloroquine. Mitochondrial morphology, ROS production and autophagy microscopy experiments were analyzed using a custom acquisition and analysis microscopy software (ImageJ). RESULTS: Exposing CRL.1790 cells to microbial challenge stimulated cells to produce several relevant biomarkers associated with inflammation and oxidative stress. Heat killed cecal contents treatment induced a 10-12 fold increase in IL-8 production by CRL.1790 cells compared to unstimulated controls at 6 and 12 h (P < 0.001). Heat killed E. coli stimulation resulted in a 4-5 fold increase in IL-8 compared to the unstimulated control cells at each time point (P < 0.001). Both heat killed E. coli and HKC stimulated robust ROS production at 6 (P < 0.001), and 12 h (P < 0.01). Mitochondrial morphologic abnormalities were detected at 6 and 12 h based on reduced mitochondrial circularity and decreased mitochondrial membrane potential, P < 0.01. Microbial stimulation also induced significant autophagy at 6 and 12 h, P < 0.01. Lastly, blocking mitochondrial ROS generation using mitochondrial specific ROS scavenger reversed microbial challenge induced mitochondrial morphologic abnormalities and autophagy. CONCLUSION: The findings from this study suggest that CRL.1790 cells may be a useful alternative to other colon cancer cell lines in studying the mechanisms of oxidative stress events associated with intestinal inflammatory disorders.

Using Oral and Colon Cancer Cells for Studying the Anticancer Properties of Antimicrobial Peptides.

Antimicrobial peptides (AMPs) are of importance in defense mechanism of many organisms and are potential candidate for treatment of infections in animals and humans. AMPs exhibit a wide range of immunomodulatory activities related to innate immunity, wound healing, and inflammation. AMPs also serve as drug delivery vectors, antitumor agents, and mitogenic agents. Here, we describe the investigation of anticancer and cytotoxic activities of antimicrobial peptides by colorimetric MTT assay using smooth muscle, dental pulp stem cell, human colon cancer cell line (SW620), and human oral squamous carcinoma cell line (HSC4).

Synthesis and Biological Evaluation of Novel Indomethacin Derivatives as Potential Anti-Colon Cancer Agents.

The molecular structure of indomethacin was used as a starting scaffold for the synthesis of 20 novel analogs and to study their effects on the proliferation of three human colon cancer cell lines, HCT-116, HT-29, and Caco-2, by MTT assay. The synthesized indomethacin analogs were characterized on the basis of IR, 1 H NMR, 13 C NMR, mass spectral data, and elemental analysis results. Cytotoxicity assay results showed that the indomethacin amide analog 2 was the most potent anticancer agent (IC50 = 0.78, 0.09, and 0.0127 µg/mL) against the three colon cancer cell lines, respectively, being more potent than the standard 5-fluorouracil (IC50 = 1.8, 0.75, and 5.45 µg/mL). Interestingly, the indomethacin oxazin analog 3 and the indomethacin amide analog 8 displayed very potent anticancer activity against the HCT-116 cell line with IC50 = 0.421 and 0.27 µg/mL, respectively, much better than the reference (IC50 = 1.8 µg/mL). Additionally, analogs 3, 4b, 11, 12c, and 13a exhibited excellent antitumor activity against Caco-2 cells, with IC50 ranging from 1.5 to 4.5 µg/mL. Furthermore, analogs 2 and 8 were additionally examined for their effect on the cell cycle of HCT-116 and HT-29 cells, respectively, using flow cytometric analysis. Analog 2 arrested the cell cycle of HT-29 cells at the S phase, while 8 was found to arrest the cell cycle of HCT-116 cells at the G0/G1 phase.

[Alpha-1 Antitrypsin Affects U0126-Induced Cytotoxicity in Colon Cancer Cell Line (HCT116)].

Alpha-1-antitrypsin (AAT), an acute phase protein, is the principal circulatory anti-protease. This multifunctional protein is encoded by the SERPINA1 gene. Although AAT was recognised as a potential tumour marker, its role in cancer biology remains unknown. Given that it has been demonstrated that AAT has an anti-apoptotic property against non-malignant cells, we aimed to investigate whether AAT affects apoptosis in a colon cancer cell line (HCT116). The presence of AAT in the HCT116 cell culture antagonized cytotoxicity of blockers of MEK1/2, PI3K/Akt pathways as well as NF-κB. The dominantly recovered cell viability was observed in the co-treatment with MEK1/2 inhibitor U0126. In addition, it was revealed that AAT almost completely abolished U0126-induced apoptosis through maintenance of the autophagy process. Our study revealed for the first time that the observed cyto-protection triggered by AAT was accompanied by sustained autophagy which opposed apoptosis. These results may contribute to understanding of the role of AAT in cancer development and evaluation of efficacy of cancer therapy.