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Background and purpose: Today, despite the abundance of chemical drugs, the use of medicinal plants is increasing; Curcumin (curcumin) is a yellow pigment obtained from the turmeric plant and is considered the effective ingredient of turmeric. This crystalline and crystallized product is widely used in Indian medicine. Recently, the beneficial effects of curcumin on stomach, colon, and mouth cancers have been proven in mice. Curcumin usually plays its role through pharmacological processes such as antioxidant, anti-inflammatory, antithrombotic, apoptotic, and hepatoprotective effects. Antioxidative, antiproliferative, and antiangiogenic properties of curcumin have been noticed in recent years. However, the low solubility of curcumin and its severe degradation have made it impossible to introduce it in clinical use. In recent years, studies and reviews have been conducted to prepare and design polymeric micelles (PMMCs) on a nanoscale to improve the cellular transport of curcumin. Green tea flavonoids, including epicatechin, have recently received attention. In in vitro research, these compounds have been proposed as antioxidants by inhibiting lipid peroxidation, trapping free radicals, and chelating metal ions, and this property is mentioned due to their unique structure (therefore, the purpose of this study is to determine the effect The synergism of curcumin and epicatechin has been carried out in the investigation of apoptosis and cell proliferation of HT29 cell lines and apoptotic factors. Materials and methods: This study is experimental. After obtaining HT29 cells from the Pasteur Institute cell bank and completing cell passage and proliferation, the cells were distributed in triplicate wells across 7 test groups and 2 control groups (positive and negative). In test groups 1, 2, and 3, three concentrations of curcumin nanomicelles (10, 20, 50 µg/ml) were incubated in triplicate. In groups 4, 5, and 6, three concentrations of epicatechin (10, 20, 50 µg/ml) were incubated for 24 hours. Test group 7 consisted of HT29 cells treated with the IC50 concentration of nanomicelle curcumin combined with the IC50 concentration of epicatechin. Group 8, the positive control, consisted of cells treated with 5FU (100 µg/ml), while group 9, the negative control, consisted of cells exposed only to the cell culture medium. After the 24-hour incubation period, an MTT assay was performed to assess cell proliferation, and Annexin flow cytometry was used to determine the level of apoptosis. Results: In this study, epicatechin and curcumin have been able to decrease cell viability of colon cancer cells (HT29) and increase the percent of apoptotic cells P

Background and purpose: Today, despite the abundance of chemical drugs, the use of medicinal plants is increasing; Curcumin (curcumin) is a yellow pigment obtained from the turmeric plant and is considered the effective ingredient of turmeric. This crystalline and crystallized product is widely used in Indian medicine. Recently, the beneficial effects of curcumin on stomach, colon, and mouth cancers have been proven in mice. Curcumin usually plays its role through pharmacological processes such as antioxidant, anti-inflammatory, antithrombotic, apoptotic, and hepatoprotective effects. Antioxidative, antiproliferative, and antiangiogenic properties of curcumin have been noticed in recent years. However, the low solubility of curcumin and its severe degradation have made it impossible to introduce it in clinical use. In recent years, studies and reviews have been conducted to prepare and design polymeric micelles (PMMCs) on a nanoscale to improve the cellular transport of curcumin. Green tea flavonoids, including epicatechin, have recently received attention. In in vitro research, these compounds have been proposed as antioxidants by inhibiting lipid peroxidation, trapping free radicals, and chelating metal ions, and this property is mentioned due to their unique structure (therefore, the purpose of this study is to determine the effect The synergism of curcumin and epicatechin has been carried out in the investigation of apoptosis and cell proliferation of HT29 cell lines and apoptotic factors. Materials and methods: This study is experimental. After obtaining HT29 cells from the Pasteur Institute cell bank and completing cell passage and proliferation, the cells were distributed in triplicate wells across 7 test groups and 2 control groups (positive and negative). In test groups 1, 2, and 3, three concentrations of curcumin nanomicelles (10, 20, 50 µg/ml) were incubated in triplicate. In groups 4, 5, and 6, three concentrations of epicatechin (10, 20, 50 µg/ml) were incubated for 24 hours. Test group 7 consisted of HT29 cells treated with the IC50 concentration of nanomicelle curcumin combined with the IC50 concentration of epicatechin. Group 8, the positive control, consisted of cells treated with 5FU (100 µg/ml), while group 9, the negative control, consisted of cells exposed only to the cell culture medium. After the 24-hour incubation period, an MTT assay was performed to assess cell proliferation, and Annexin flow cytometry was used to determine the level of apoptosis. Results: In this study, epicatechin and curcumin have been able to decrease cell viability of colon cancer cells (HT29) and increase the percent of apoptotic cells P<0.05, which was in a dose-dependent manner, also the combination of these two drugs caused a synergistic effect in high dose P<0.05. Epicatechin in concentrations of 20, 10 μg/mL, 50, and 100 has a significant difference compared to the negative control group with P<0.05, which has caused a decrease in cell viability in colon cancer cells with increasing dose. Curcumin is the group Nanomicelle with epicatechin at IC50 concentration compared to the positive control group of 5-FU in HT-29 cell line has greatly reduced cell viability of HT-29 cell line showed that curcumin and epicatechin nano micelles had a synergistic effect. The results of cell apoptosis have shown that curcumin and epicatechin nano micelle groups at IC50 concentrations had a synergistic effect. Conclusion: epicatechin and curcumin have been able to increase the death rate of colon cancer cells (HT29) by increasing apoptosis, which indicates the anti-cancer effects of these compounds. So it can suggest these compounds in colon cancer therapeutics. [ABSTRACT FROM AUTHOR]

Background and Objectives: Colorectal cancer (CRC) is a common type of cancer that has a high death rate and is becoming more common in developed countries. Currently, there are several treatment options available for CRC patients, and clinical trials are being conducted to improve conventional therapies. This study investigates the combined impact of Bacillus coagulans (B.C) and Newcastle disease virus (NDV) on the growth of human colorectal adenocarcinoma cells (HT29 cell line). Materials and Methods: The HT29 cell line was cultured under controlled laboratory conditions. They were treated with Fluorouracil (5-FU), NDV, and B.C., after which various assessments were conducted to determine the effects of these treatments. These assessments included MTT assay for cytotoxicity, evaluation of cell viability, and measurement of caspase 8 and 9 activity levels. The significance of the data was determined at a threshold of P<0.05 following analysis. Results: The usage of NDV and B.C significantly increased cell death and reduced cell growth in the HT29 cell line, when compared to the control group. Moreover, the combined application of NDV and B.C along with 5-FU exhibited a synergistic effect in decreasing the proliferation of HT29 cells. Additionally, the results indicated that intrinsic apoptosis pathway was activated by B.C and NDV. Conclusion: It appears that utilizing oncolytic viruses (OV) and bacteria in conjunction with chemotherapy drugs could potentially aid in reducing the growth of colorectal cancer cells. However, further research is necessary, including animal studies, to confirm the efficacy of this treatment method. [ABSTRACT FROM AUTHOR]

Background and Objectives: Colorectal cancer (CRC) is a common type of cancer that has a high death rate and is becoming more common in developed countries. Currently, there are several treatment options available for CRC patients, and clinical trials are being conducted to improve conventional therapies. This study investigates the combined impact of Bacillus coagulans (B.C) and Newcastle disease virus (NDV) on the growth of human colorectal adenocarcinoma cells (HT29 cell line). Materials and Methods: The HT29 cell line was cultured under controlled laboratory conditions. They were treated with Fluorouracil (5-FU), NDV, and B.C., after which various assessments were conducted to determine the effects of these treatments. These assessments included MTT assay for cytotoxicity, evaluation of cell viability, and measurement of caspase 8 and 9 activity levels. The significance of the data was determined at a threshold of P

Arabinoxylan has prebiotic properties, as it is able to resist digestion in the small intestine and undergoes fermentation in the large intestine. In this work, arabinoxylan was extracted from corn fiber using an alkaline solution and further purified with membrane processing. It was found that the extracts were mainly composed of xylose (50–52%), arabinose (37–39%), galactose (9%) and glucose (1–4%), with an A/X ratio of 0.72–0.77. All the extracts were composed of phenolic compounds, including ferulic acid derivatives such as dimers, trimers and tetramers. The purified extract had a lower concentration of ferulic and p-coumaric acid (0.004 and 0.02 mg/mgdry_weight, respectively) when compared to raw extract (19.30 and 2.74 mg/mgdry_weight, respectively). The same effect was observed for the antioxidant activity, with purified extracts having a lower value (0.17 ± 0.02 µmol TEAC/mg) when compared to the raw extract (2.20 ± 0.35 µmol TEAC/mg). The purified extract showed a greater antiproliferative effect against the HT29 cell line with EC50 = 0.12 ± 0.02 mg/mL when compared to the raw extract (EC50 = 5.60 ± 1.6 mg/mL). Both raw and purified extracts did not show any cytotoxicity to the Caco-2 cell line in the maximum concentration tested (10 mg/mL). [ABSTRACT FROM AUTHOR]

Purpose: Colorectal cancer (CRC) is one of the most common and fatal malignancies in humans, still leading to serious morbidity and mortality. We here aimed to investigate the effects of flavonoid apigenin, which is considered to have anti-tumoral activity on CRC with high epidemiological prevalence, on cell proliferation and cell survivals, and the positive and negative dose-dependent effects of genetic or mutational alterations in SWH pathway components on HT29 CRC cell lines. Methods: Human colon cancer cell lines HT-29 were commercially available. In each flask, 5 groups were formed, each of which consists of 5,000 cells for different dose groups and the cells were plated. After a 24 and 48 h incubation period, cytotoxicity values were measured by MTT assay and gene expression was assessed by real-time polymerase chain reaction (PCR) analysis method. Results: Application of 12.5 and 25 nM of apigenin significantly increased cell death in HT29 cell lines. LATS1, STK3 and TP53 gene expression decreased in the same dose groups compared to control and other groups. Conclusion: It has been concluded that TP53 gene is strongly correlated with LATS1 and STK3 genes among the SWH pathway factors in the progression of CRC and could be used as an important marker for early detection of malignant transmission. In addition, it may be effective in CRC cases especially when 25 nM of apigenin applies for therapeutic purpose.

Purpose: Colorectal cancer (CRC) is one of the most common and fatal malignancies in humans, still leading to serious morbidity and mortality. We here aimed to investigate the effects of flavonoid apigenin, which is considered to have anti-tumoral activity on CRC with high epidemiological prevalence, on cell proliferation and cell survivals, and the positive and negative dose-dependent effects of genetic or mutational alterations in SWH pathway components on HT29 CRC cell lines. Methods: Human colon cancer cell lines HT-29 were commercially available. In each flask, 5 groups were formed, each of which consists of 5,000 cells for different dose groups and the cells were plated. After a 24 and 48 h incubation period, cytotoxicity values were measured by MTT assay and gene expression was assessed by real-time polymerase chain reaction (PCR) analysis method. Results: Application of 12.5 and 25 nM of apigenin significantly increased cell death in HT29 cell lines. LATS1, STK3 and TP53 gene expression decreased in the same dose groups compared to control and other groups. Conclusion: It has been concluded that TP53 gene is strongly correlated with LATS1 and STK3 genes among the SWH pathway factors in the progression of CRC and could be used as an important marker for early detection of malignant transmission. In addition, it may be effective in CRC cases especially when 25 nM of apigenin applies for therapeutic purpose. [ABSTRACT FROM AUTHOR]

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Arabinoxylan has prebiotic properties, as it is able to resist digestion in the small intestine and undergoes fermentation in the large intestine. In this work, arabinoxylan was extracted from corn fiber using an alkaline solution and further purified with membrane processing. It was found that the extracts were mainly composed of xylose (50–52%), arabinose (37–39%), galactose (9%) and glucose (1–4%), with an A/X ratio of 0.72–0.77. All the extracts were composed of phenolic compounds, including ferulic acid derivatives such as dimers, trimers and tetramers. The purified extract had a lower concentration of ferulic and p-coumaric acid (0.004 and 0.02 mg/mgdry_weight, respectively) when compared to raw extract (19.30 and 2.74 mg/mgdry_weight, respectively). The same effect was observed for the antioxidant activity, with purified extracts having a lower value (0.17 ± 0.02 µmol TEAC/mg) when compared to the raw extract (2.20 ± 0.35 µmol TEAC/mg). The purified extract showed a greater antiproliferative effect against the HT29 cell line with EC50 = 0.12 ± 0.02 mg/mL when compared to the raw extract (EC50 = 5.60 ± 1.6 mg/mL). Both raw and purified extracts did not show any cytotoxicity to the Caco-2 cell line in the maximum concentration tested (10 mg/mL).

In this study, chemical composition (gas chromatography-mass spectroscopy), chemical structure (fourier transform infrared spectroscopy) and antioxidant potential (β-carotene bleaching assay and DPPH/ABTS-radical scavenging activity tests) of Syzygium aromaticum essential oil (SAEO) were evaluated. Eugenol (75.11%) was found to be the major compound of SAEO. Eugenol, as the main chemical constituent of SAEO, showed its signature peaks in the wavenumber range of 720 – 1250 cm-1, ascribing to the C=C region. The antimicrobial activity of SAEO on Escherichia coli, Staphylococcus aureus, Listeria innocua and Pseudomonas aeruginosa were evaluated. The scanning electron microscopy (SEM) was then applied to unravel the antibacterial mechanism of SAEO on E. coli as the most resistant strain and L. innocua as the most sensitive strain. The MTT assay was also used to investigate the cytotoxicity effect of SAEO on human colonic cancer cell lines (HT29 cell line) and the highest cytotoxic effect was observed at 200 mg.mL-1 concentration of SAEO. The SEM micrographs revealed that the SAEO treatment was able to manifestly increase the cell permeabilization and membrane integrity disruption. This means that the entirety of the cell membranes was remarkably affected by the essential oil, which could lead to cytoplasm secretion and subsequent cell death. The data strongly suggest that SAEO had a potential antioxidant, antimicrobial and cytotoxicity activity.

Background: Colon cancer is one of the most common cancers in the world, and efforts toward its treatment have not been completely successful. In recent years, more attention has been focused on herbal medicine (HM) due to their anticancer and cytotoxic properties. This study investigated the anticancer and antioxidant effects of Artemisia aucheri (A. aucheri) Boiss extract against HT29 colon cancer cells compared with HEK239 natural cells. Methods: This study was performed on human HT29 colon cancer cells. Various doses of 0, 10, 100, 500, and 1000 μg/ml of A. aucheri extract were subjected to cells at specified time intervals. After treatment, the trypan blue test was employed to determine the viability of the cells. MTT and annexin tests were used to determine cell viability and the apoptosis induced by the extract. Malondialdehyde (MDA) testing was applied to investigate the antioxidant properties of the extract on fatty acids. Data analysis was performed using SPSS version 22. One-way ANOVA and paired comparison tests were employed for data analysis. Results: The highest cytotoxicity effect of A. aucheri extract was observed at 1000 μg/ml (80.63 ± 3.66) being dose-dependent compared with the control in both cell lines (p < 0.001). Additionally, the survival rate of HT29 (IC50 = 57.88 μg/ml) and HEK (IC50 = 295 μg/ml) cancer cells decreased with increasing concentration of A. aucheri (the lowest cell viability was at 1000 μg/ml). Furthermore, the induction of membrane lipid peroxidation was significantly higher in HT29 compared with the control (p < 0.001). Another cytotoxic mechanism for the extract was the induction of apoptosis being significantly higher in HT29 colon cancer cells compared with the control group (p < 0.001). Conclusion: Cytotoxic effects of A. aucheri extract were dose-dependent. This HM exerted cytotoxic effects against HT29 cells through the induction of membrane lipid peroxidation and apoptosis. [ABSTRACT FROM AUTHOR]

The effects of a control bread (CB) and a Gluten Friendly™ bread (GFB) on intestinal epithelium mucus production and barrier function in healthy human mucus-secreting goblet cells HT-29-16E were investigated. Mucus production in cells exposed to digested breads (GFB and CB) was preliminarily investigated using staining techniques, Periodic Acid-Schiff (PAS) and Alcian blue (AB), and MUC2 and MUC3 were also quantified by ELISA assay. The barrier function of the cell monolayer was evaluated by trans-epithelial electrical resistance (TEER) measurements. GFB increased the secretion of mucins, expressed as the level of PAS and AB staining in comparison with the control. MUC3 levels were not affected, whereas higher MUC2 concentrations (P

Introduction: Niosome is a lipid carrier that can resolves some of the challenges facing the delivery of herbal medicines to tissues. The aim of the present study is to fabricate and evaluate the physiochemical properties of niosomal carriers containing Hedera helix extracts and evaluation of its toxicity to the HT29 cell line of colon cancer. Methods: Two niosomal formulations F0 (30% cholesterol and 70% tween60) and F1 (30% cholesterol and 70% spin60) containing extract of Hedera hilix were made using the thin film method. Then, according to the amount of drug loading, one of the formulations was selected. The pattern of drug release under normal and cancerous cell conditions, size and surface charge of the nanoparticles (using DLS) and the appearance of the nanoparticles (using SEM) of the selected formulation were investigated. Finally, the toxicity of the niosomal system containing extracts and free extracts on HT29 cell line was evaluated by MTT assay and niosomal system entry into HT29 cells was evaluated by system and cell staining. Results: Extract loading percentage, size and zeta potential for the selected formulation (F1) containing the extract were 95/43±2/43%, 132/5nm and -41/47±2/69mV, respectively. Release of the extract from the niosomal system is slow within 48 hours in normal and cancerous cell conditions. The appearance of the nanoparticles was smooth and spherical and extract niosomal had more toxicity to HT29 cell line colon cancer than free extract. Conclusion: The niosomal formulation of this study can be recommended for further research in colon cancer with respect to its physicochemical properties. [ABSTRACT FROM AUTHOR]

Introduction: Bifunctional radiosensitizer agents in which nitroaromatic moieties are attached through a linker to antineoplastic moieties have demonstrated higher cytotoxicity and radiosensitizer effects than the corresponding counterparts. This study was conducted to investigate the cytotoxicity and radiosensitizer activities of 2, 4-dinitrobenzene as a radiosensitizer moiety which connected to α, β unsaturated aryl ketones against the radioresistant human HT29 colon cancer cells. Methods: A series of bifunctional radiosensitizer derivatives that are composed of electron-affinic 2, 4-dinitrophenyl moiety and thiol reactive unsaturated conjugated ketones were prepared. The designed compounds were synthesized by the reaction of the corresponding 2, 4-dinitrobenzaldehyde, cyclohexanone and different aryl aldehydes. The cytotoxicity and radiosensitizer activity of the tested compounds were examined against HT29 colon cancer cells under aerobic condition. The IC50 value of the tested compounds and percent of survival cells were analyzed by the MTT assay. The clonogenic assay was used to assess the cell viability following treatment with the tested compounds with or without the combination of radiation. Results: This approach demonstrated that the tested compounds at the concentrations utilized have little or no cytotoxicity towards the radioresistant HT29 cell line but have great cytotoxicity and radiosensitizer activity when combined with irradiation. Conclusion: The novel bifunctional unsaturated conjugated aryl ketones which are thiol alkylators found to exhibit radiosensitivity activity. Consequently, these new developed compounds should be evaluated further to assess their potential efficacy with radiotherapy to combat malignancies in a pre-clinical animal model. [ABSTRACT FROM AUTHOR]

In this study, chemical composition (gas chromatography-mass spectroscopy), chemical structure (fourier transform infrared spectroscopy) and antioxidant potential (β-carotene bleaching assay and DPPH/ABTS-radical scavenging activity tests) of Syzygium aromaticum essential oil (SAEO) were evaluated. Eugenol (75.11%) was found to be the major compound of SAEO. Eugenol, as the main chemical constituent of SAEO, showed its signature peaks in the wavenumber range of 720 - 1250 cm-1, ascribing to the C=C region. The antimicrobial activity of SAEO on Escherichia coli, Staphylococcus aureus, Listeria innocua and Pseudomonas aeruginosa were evaluated. The scanning electron microscopy (SEM) was then applied to unravel the antibacterial mechanism of SAEO on E. coli as the most resistant strain and L. innocua as the most sensitive strain. The MTT assay was also used to investigate the cytotoxicity effect of SAEO on human colonic cancer cell lines (HT29 cell line) and the highest cytotoxic effect was observed at 200 mg.mL-1 concentration of SAEO. The SEM micrographs revealed that the SAEO treatment was able to manifestly increase the cell permeabilization and membrane integrity disruption. This means that the entirety of the cell membranes was remarkably affected by the essential oil, which could lead to cytoplasm secretion and subsequent cell death. The data strongly suggest that SAEO had a potential antioxidant, antimicrobial and cytotoxicity activity. [ABSTRACT FROM AUTHOR]

Graphical abstract Highlights • Temperature-treated Gluten Friendly™ bread (GFB) was studied. • GFB and control bread (CB) are not cytotoxic for the HT29-16E and HT29-19A cells. • GFB increases the secretion of mucins in human mucus-secreting goblet cells. • GFB increases significantly the level of MUC2 with respect to CB and control. • GFB treated cells have significantly higher TEER values than CB and control cells. Abstract The effects of a control bread (CB) and a Gluten Friendly™ bread (GFB) on intestinal epithelium mucus production and barrier function in healthy human mucus-secreting goblet cells HT-29-16E were investigated. Mucus production in cells exposed to digested breads (GFB and CB) was preliminarily investigated using staining techniques, Periodic Acid-Schiff (PAS) and Alcian blue (AB), and MUC2 and MUC3 were also quantified by ELISA assay. The barrier function of the cell monolayer was evaluated by trans-epithelial electrical resistance (TEER) measurements. GFB increased the secretion of mucins, expressed as the level of PAS and AB staining in comparison with the control. MUC3 levels were not affected, whereas higher MUC2 concentrations (P < 0.01) were found on cells treated with GFB compared to the control. Additionally, significantly higher TEER values were observed after treatment with both CB and GFB in comparison with the control (P < 0.01), with GFB having a significantly higher effect than CB (P < 0.01). [ABSTRACT FROM AUTHOR]

Fruits and vegetables are a good source of potentially biologically active compounds. Their regular consumption in the human diet can help reduce the risk of developing chronic diseases such as cardiovascular diseases and cancer. Plants produce additional chemical substances when subject to abiotic stress or infected by microorganisms. The phytochemical profile of spinach leaves (Spinacia oleracea L.), which is a vegetable with widely recognized health-promoting activity, has been affected by applying root hypoxic and re-oxygenation stress during plant growth. Leaf juice at different sampling times has been subject to liquid chromatography mass spectrometry (LC-MSn) analysis and tested on the human colorectal adenocarcinoma cell line HT29 by using the Comet assay. The cells were previously treated with H2O2 to simulate the presence of an oxidative stress (as in colon cancer condition) and the leaf juice application resulted in a significant antioxidant and protective in vitro effect. The duration of the hypoxic/re-oxygenation stress imposed on the plant reflects the antioxidant leaf juice content. After hypoxic stress (24 h) and reoxygenation (2 h), we show a decrease (50%) of the relative abundance of the principal identified antioxidant molecules but a higher antioxidant activity of the spinach juice on HT29 cells (20%). Data shows a complex relation between plant growing conditions and the modulation of secondary metabolites content in leaf juice that results in different chemo-protective activities in colon cancer cells.

New unsymmetrical mesoporphyrinic complexes, namely 5-(4-hydroxyphenyl)-10,15,20–tris-(4-carboxymethylphenyl)–21,23-Zn(II)-porphine and 5-(4-hydroxyphenyl)-10,15,20–tris-(4-carboxymethylphenyl)–21,23-Cu(II)-porphine, were synthesized using a microwave irradiation method. The structures of the porphyrinic complexes were confirmed using FT-IR, UV–Vis, EPR and NMR spectral data. The spectral absorption and emission properties of the porphyrinic complexes were studied in organic solvents of different polarities and the influence of solvent polarity on the wavelengths of the absorbance and fluorescence band maxima is described. The cytotoxicity evaluation of the porphyrinic complexes was performed on human colon adenocarcinoma cell line HT29 for different doses and incubation times. The obtained result indicates a lack of or low toxicity for both compounds, thus recommending them for further testing in light activation protocols.

The bioactive compounds of brown rice exhibit many beneficial health effects, ranging from antioxidant to cytotoxic activities. Pulsed Electric Field (PEF) pretreatment can significantly enhance their extraction, through the induction of the electro-permeabilization of the cell membranes. This paper aims to demonstrate that PEF-assisted extraction of brown rice enables not only enhanced yields of antioxidant compounds, such as γ-oryzanol, polyphenols and phenolic acids, and of saturated and unsaturated fatty acids, but also increased cytotoxic effects on cancer cells. Initially, the PEF-assisted extraction conditions have been defined by the assessment of the cell permeabilization index via impedance measurements and the DPPH antioxidant activity. Subsequently, the biological effects of PEF have been evaluated on the cytotoxicity and anti-inflammatory properties against human colon cancer cell line HT29. The results show that PEF-assisted extraction, enhancing the yield of bioactive compounds, with respect to untreated extracts, significantly promotes their antioxidant activity, which is correlated with an increased HT29 cells cytotoxicity. In addition, PEF extracts of brown rice substantially inhibit also gene expression and interleukin production in colon cancer cells, suggesting their exploitation as natural anti-inflammatory agents. The integration of PEF pretreatment in the solvent extraction process of bioactives from brown rice appears, therefore, as a promising practice to significantly enhance their biological activity. [ABSTRACT FROM AUTHOR]

Human exposure to Bisphenol A (BPA) results mainly from ingestion of food and beverages. Information regarding BPA effects on colon cancer, one of the major causes of death in developed countries, is still scarce. Likewise, little is known about BPA drug interactions although its potential role in doxorubicin (DOX) chemoresistance has been suggested. This study aims to assess potential interactions between BPA and DOX on HT29 colon cancer cells. HT29 cell response was evaluated after exposure to BPA, DOX, or co-exposure to both chemicals. Transcriptional analysis of several cancer-associated genes ( c-fos , AURKA , p21 , bcl-xl and CLU ) shows that BPA exposure induces slight up-regulation exclusively of bcl-xl without affecting cell viability. On the other hand, a sub-therapeutic DOX concentration (40 nM) results in highly altered c-fos , bcl-xl , and CLU transcript levels, and this is not affected by co-exposure with BPA. Conversely, DOX at a therapeutic concentration (4 μM) results in distinct and very severe transcriptional alterations of c-fos , AURKA , p21 and CLU that are counteracted by co-exposure with BPA resulting in transcript levels similar to those of control. Co-exposure with BPA slightly decreases apoptosis in relation to DOX 4 μM alone without affecting DOX-induced loss of cell viability. These results suggest that BPA exposure can influence chemotherapy outcomes and therefore emphasize the necessity of a better understanding of BPA interactions with chemotherapeutic agents in the context of risk assessment. [ABSTRACT FROM AUTHOR]

Submicrometric lipid-based carriers were developed to encapsulate curcumin and deliver it to intestinal epithelial cells. A lipid matrix comprising monoolein, sunflower oil and water at weight ratio 1:1:1 was selected, upon screening of different combinations of amphiphilic molecules, vegetable oils and water, because of its high encapsulations efficiency of curcumin, retained over time and relatively lower content of amphiphilic molecules. Upon dispersion in aqueous phase, the carriers were stabilized by: (a) whey protein isolates (WPI), alone and (b) in combination with modified starch (WPI-MS), or by (c) polysorbate 20 (T20). Whereas T20-stabilized systems exhibited extremely fine particles (120 nm), WPI and WPI-MS stabilized carriers were characterized by a significantly larger mean particle size (270 nm). The thicker macromolecular layer of WPI and WPI-MS enabled better (a) physical stability, (b) controlled shell degradation during simulated digestion, and (c) curcumin bioaccessibility targeted at the intestinal digestion phase than T20-systems. However, uptake studies in HT29 cell lines, simulating intestinal epithelial cells, showed that WPI and WPI-MS carriers exhibited after 24 h a lower relative uptake than T20-stabilized systems (about 60% and 80%, respectively), as a consequence of smaller size and higher cell adherence of T20 carriers to the cell membrane. [ABSTRACT FROM AUTHOR]

2,3'-Biindole (2) was synthesized via bromination of indole (1) with molecular bromine and underwent simultaneous dimerization. Antiproliferative and cytotoxic activity of 2 was investigated in vitro on C6 (rat brain tumor), HeLa (human cervix carcinoma), and HT29 (human colon carcinoma) cells lines by using BrdU cell proliferation ELISA and lactate dehydrogenase (LDH) assays. In contrast to 5-fluorouracil (5-FU), 2,3'-biindole (2) significantly inhibited proliferation of HeLa and HT29 cell lines. According to LDH assay, the cytotoxicity of compound 2 was low on HT29 cell lines and high on HeLa and C6 cell lines. Moreover, 2 did not cause any DNA laddering on the DNA of tested cells; therefore, it is suggested that the mechanism of action of this compound may not involve apoptosis. In addition, 2 inhibited relaxation of supercoiled plasmid DNA by topoisomerase activity. Results of the present study indicates that biindole (2) may have promising anticancer and antitopoisomerase potential with an unknown mechanism of action. [ABSTRACT FROM AUTHOR]

Aim of the study was to compare radiobiological effects of multiple vs. single low-dose pre-irradiation on the HT29 cell line. This regime is designed to be as similar as possible to fractionated tumour radiotherapy treatment, and to provide data on radiobiological effects on human tumour cells. Material and methods: The cell line used in the study was HT29 (human colorectal adenocarcinoma, American Type Culture Collection HTB-38™). Also, for comparison, the MRC5 cell line (human foetal lung fibroblasts, American Type Culture Collection CCL 171) was used. Four-day treatment in a 4 x 2 Gy regime was performed. Cell viability was evaluated by tetrazolium colorimetric MTT assay. Results: Multiple low-dose pre-irradiation induced a stronger radioadaptive response compared to single low-dose application in the HT29 cell line. Multiple pre-irradiation with 0.03 Gy and 0.05 Gy caused radioadaptive effects, while in both single and multiple low-dose pre-irradiation regimes 0.07 Gy led to radiosensitivity. Radiobiological effects induced in the HT29 cell line by low-dose pre-irradiation were evidently weak during the treatment time, because a single low-dose applied only on the first day gave no radioadaptive effects. In the MRC5 cell line different effects were registered, since radioadaptive response has not been observed after multiple or single pre-irradiation. Conclusions: The obtained data are interesting, especially for the possible application of low-dose pre-irradiation in radiotherapy. [ABSTRACT FROM AUTHOR]

Abstract: During the last years, special interest has been focused on the recovery of bioactive compounds from food industry wastes in an effort to find added value applications for residues. In this work, Opuntia spp. residues from fruit juice production were explored as potential sources of natural chemotherapeutic ingredients towards colon cancer. Hydroalcoholic extraction and adsorption separation processes (with Amberlite XAD16 resin) were used to produce natural extracts from residues of Opuntia ficus-indica and Opuntia robusta and their antiproliferative effect was evaluated in human colon carcinoma HT29 cell line. Our results showed that the natural extracts efficiently inhibited cancer cell growth and induced cell cycle arrest in different checkpoints—G1, G2/M and S. The phytochemical constituents presented in the samples, namely betacyanins, flavonoids (isorhamnetin derivatives) and phenolic acids (ferulic acid) were identified as the main responsible compounds for the cell cycle arrest. Moreover, the extracts that presented the lowest effective dose values in terms of polyphenols (ED50 value<0.5mgGAE/mL) showed to increase intracellular ROS accumulation in HT29 cells, suggesting that cancer cell death may be induced by the pro-oxidant effect of these compounds. [Copyright &y& Elsevier]

Purpose: To develop PEGylated multi-walled carbon nanotubes as a sustained release drug delivery system. Methods: Oxaliplatin was incorporated into inner cavity of PEGylated multi-walled carbon nanotubes (MWCNT-PEG) using nano-extraction. Oxaliplatin release rates from MWCNT-PEG-Oxaliplatin were investigated using dialysis tubing. Cytotoxicity of oxaliplatin, MWCNT-Oxaliplatin and MWCNT-PEG-Oxaliplatin were evaluated in HT29 cell by MTT assay, Pt-DNA adducts formation, γ-H2AX formation and cell apoptosis assay. Results: Loading of oxaliplatin into MWCNT-PEG was ~43.6%. Sustained release occurred to MWCNT-PEG-Oxaliplatin, with only 34% of oxaliplatin released into medium within 6 h. In MTT assay, MWCNT-PEG-Oxaliplatin showed slightly decreased cytotoxic effect when cell viability was assessed at 12 and 24 h. A drastic increase of cytotoxicity was found when cell viability was assessed at 48 and 96 h. Pt-DNA adducts formation, γ-H2AX formation and cell apoptosis assay results showed the same trend as the MTT assay, suggesting sustained-release for MWCNT-Oxaliplatin and MWCNT-PEG-Oxaliplatin formulations. Conclusions: PEGylated multi-walled carbon nanotubes can be used as sustained release drug delivery system, thus remarkably improving cytotoxicity of oxaliplatin on HT-29 cells. [ABSTRACT FROM AUTHOR]

Abstract: The aim of the present study was to reduce 5-FU side effects by targeted nanostructured lipid carriers (NLCs) to LDL receptors that are over expressed in colorectal carcinoma and also use of a new synthesized conjugate of retinoic acid as a cytotoxic agent. Fatty acyl amide derivative of retinoic acid was synthesized by its conjugation to octadecylamine with the expectation to improve its loading capacity in NLCs of 5-FU. The NLCs were prepared by an emulsification-solvent evaporation method using cholesterol and cholesteryl stearate. Physical properties and drug release were studied in NLCs. The cytotoxicity of NLCs loaded with 5-FU and retinoic acid conjugate was studied on colon cancer cells (HT29) using MTT assay. To confirm that drug targeting has been done through LDL receptors, APO-E was omitted from the cell culture and the MTT assay was repeated. FTIR and 1H NMR spectra confirmed successful production of the conjugate. Results showed the IC50 of free 5-FU was about 7.6 μM while in comparable concentration, the cytotoxicity of 5-FU loaded in NLCs containing the retinoic acid conjugate was nearly 2 fold of NLCs just loaded with 5-FU and more than 5 fold of free 5-FU. The retinoic acid conjugate loaded NLCs prepared by cholesterol can target LDL receptors of HT29 cells and seems promising in reducing 5-FU dose in colorectal cancer. [Copyright &y& Elsevier]

Apigenin (4',5,7-trihydroxyfiavone) is one of the leading components supporting targeted treatment options. We explored the cytotoxic and apoptotic effects of various doses of apigenin administered alone and together with 5-fluorouracil (5-FU)--a chemotherapeutic agent with high cytotoxicity--for different incubation periods, on morphologic, DNA, RNA (messenger RNA [mRNA]), and protein levels on the p53 mutant HT29 human colon adenocarcinoma cell line. Treatment with apigenin alone for a 72-hour incubation at 90-µM dose resulted in an apoptotic percentage of 24.92% (P = .001). A higher percentage (29.13%) was observed after treatment with the same dose of apigenin plus 5-FU for the same incubation period (P=.001). These results were confirmed as mRNA and protein expression levels of caspase-3 increased 2.567-fold and mRNA expression levels of caspase-8 increased 3.689-fold compared with the control group. On the other hand, mRNA expression levels of mammalian target of rapamycin (mTOR) and cyclin D1 (CCND1) decreased by 0.423-fold and 0.231-fold, respectively. To our knowledge this is the first study showing that treatment with apigenin alone results in cell cycle arrest through activation of caspase cascade and stimulation of apoptosis in HT29 cells. It also shows that use of apigenin plus 5-FU further increases this effect. This study draws attention to the probable clinical effectiveness of apigenin plus a chemotherapeutic agent with high cytotoxicity. It also highlights the induction of desirable apoptotic effects by targeting the caspase cascade pathway through administration of reduced doses for shorter incubation periods. [ABSTRACT FROM AUTHOR]

New unsymmetrical mesoporphyrinic complexes, namely 5-(4-hydroxyphenyl)-10,15,20-tris-(4-carboxymethylphenyl)-21,23-Zn(II)-porphine and 5-(4-hydroxyphenyl)-10,15,20-tris-(4-carboxymethylphenyl)-21,23-Cu(II)-porphine, were synthesized using a microwave irradiation method. The structures of the porphyrinic complexes were confirmed using FT-IR, UV-Vis, EPR and NMR spectral data. The spectral absorption and emission properties of the porphyrinic complexes were studied in organic solvents of different polarities and the influence of solvent polarity on the wavelengths of the absorbance and fluorescence band maxima is described. The cytotoxicity evaluation of the porphyrinic complexes was performed on human colon adenocarcinoma cell line HT29 for different doses and incubation times. The obtained result indicates a lack of or low toxicity for both compounds, thus recommending them for further testing in light activation protocols. [ABSTRACT FROM AUTHOR]

Background: Photodynamic therapy (PDT) involves killing of diseased cells by excitation of photosensitizer chemicals with high-energy light to produce cytotoxic oxygen species from surrounding dissolved oxygen. However, poor tissue penetration of high-energy light and hydrophobic photosensitizers limits the effectiveness to superficial pathologies. Upconversion phosphor nanoparticles convert low-energy radiation to higher-energy emissions. Aim: To create upconverting 'nanotransducers' to enable PDT in deep tissues. Results: Monodisperse, 50 nm PEI/NaYF4:Yb3+,Er3+ nanoparticles producing green/red emission on near-infrared (NIR) excitation were targeted to folate receptors on human colon cancer cells and imaged with high signal-to-background ratio. It was demonstrated that these particles could be excited after deep intramuscular injection in rats. On NIR excitation, the particles, modified with zinc phthalocyanin photosensitizer, released singlet oxygen and, after targeted binding to cancer cells, resulted in significant cell destruction. Conclusion: Potential clinical use of these nanoparticles includes imaging and PDT of cancer in deep tissues. [ABSTRACT FROM AUTHOR]

Abstract: The application of reversed-phase high-pressure liquid chromatography under gradient conditions and electrospray ion trap mass spectrometry (LC–ESI–MS) to the analysis of global modification levels of core histones is described. The optimised LC–ESI–MS method was applied for the first time to the characterisation of histones extracted from HT29, a human colon cancer cell line. Eight histones (H1-1, H1-2, H2A-1, H2A-2, H2B, H3-1, H3-2, H4) were separated on a C4 stationary phase with complete resolution, never reached in previous HPLC–MS methods, by using a gradient elution with the combined presence of heptafluorobutyric acid and formic acid as acidic modifiers in the mobile phase. Heptafluorobutyric acid was found to improve selectivity, whereas the presence of formic acid decreased ion suppression. Histones eluted from the column were detected with an ion trap mass spectrometer with an electrospray source. The peak averaged mass spectra were reconstructed by Mag Tran 1.0 software and the mass of the various isoforms of histones were derived. Method validation was conducted by performing the same sample analysis by coupling LC–ESI to a quadrupole-time-of-flight mass spectrometer (Q-TOF). The number of histone forms and their mass were found to differ not significantly from those obtained by ion trap mass spectrometer. Also the relative modifications abundance within the same histone type was found following the same trend as the two mass analysers. This method was then applied to the characterisation of changes in histone modification in HT29, never analysed by LC–MS before, treated with histone deacetylase inhibitors such as valproate and sodium butyrate, also used in preclinical trials as anticancer drugs. In particular, both the inhibitors produced a significant increase in H4 histone acetylated forms: 89% increase of the diacetyl dimethyl H4 form was observed with 1mM valproate supplementation, whereas 5mM butyrate led to a 68% increase of the same form. Triacetyl monomethyl H4 (11377Da) and triacetyl dimethyl H4 (11390Da) were found only in cells treated with butyrate. Selective changes of H3 histone were detected with butyrate, in agreement with recently reported western blotting studies. Modifications in the H2A histone degree of acetylation were revealed by treatment of the cells with butyrate (H2A-1, H2A-2) and valproate (H2A-2). The results of the proposed methodology confirmed that inhibition of histone deacetylases caused histone hyperacetylation, responsible for decondensation and reorganization of interphase dynamic chromatin. This method resulted in selective and sensitive method to monitor variations in the acetylation and methylation state of histones after treatment of HT29 with inhibitors, and is therefore suitable for further application in new drug discovery for tumour therapy. [Copyright &y& Elsevier]

Due to ingestion of contaminated foods, the human gastrointestinal tract is the most likely site of exposure to microplastics (MPs) with gut barrier dysfunction and intestinal inflammation. Aimed to assess the effects induced by MPs with different granulometry (polystyrene (PS) 3 and 10 µm), we performed an in vitro study by using the human intestinal cell line HT29. As a novelty, we assessed the sub-chronic exposure extending the treatment up to 48 days simulating the in vivo situation. In the range of 100-1600 particles mL -1 , both the PS suspensions had moderate cytotoxicity after 24 h with percentages of mortality between 6.7 and 21.6 for the 10 µm and 6.1 and 29.6 for the 3 µm PS. Microscopic observation highlighted a more pronounced lysosomal membrane permeabilization in HT29 exposed to PS 3µm. Reactive oxygen species production was higher in cells exposed to PS 10 µm, but sub-chronic exposure highlighted the ability of the cells to partially neutralize this effect. Comet-assay confirmed the temporary oxidative damage that was PS-induced. Overall, considering the very fast turnover of intestinal cells, the increase in cell mortality, equal to 25% and 11% for 3 and 10 µm PS-MPs for each time point, could trigger intestinal disorders due to prolonged exposure.

Fruits and vegetables are a good source of potentially biologically active compounds. Their regular consumption in the human diet can help reduce the risk of developing chronic diseases such as cardiovascular diseases and cancer. Plants produce additional chemical substances when subject to abiotic stress or infected by microorganisms. The phytochemical profile of spinach leaves (Spinacia oleracea L.), which is a vegetable with widely recognized health-promoting activity, has been affected by applying root hypoxic and re-oxygenation stress during plant growth. Leaf juice at different sampling times has been subject to liquid chromatography mass spectrometry (LC-MSn) analysis and tested on the human colorectal adenocarcinoma cell line HT29 by using the Comet assay. The cells were previously treated with H2O2 to simulate the presence of an oxidative stress (as in colon cancer condition) and the leaf juice application resulted in a significant antioxidant and protective in vitro effect. The duration of the hypoxic/re-oxygenation stress imposed on the plant reflects the antioxidant leaf juice content. After hypoxic stress (24 h) and reoxygenation (2 h), we show a decrease (50%) of the relative abundance of the principal identified antioxidant molecules but a higher antioxidant activity of the spinach juice on HT29 cells (20%). Data shows a complex relation between plant growing conditions and the modulation of secondary metabolites content in leaf juice that results in different chemo-protective activities in colon cancer cells. [ABSTRACT FROM AUTHOR]

Background: Gymnema sylvestre is an ethno-pharmacologically important medicinal plant used in many polyherbal formulations for its potential health benefits. Silver nanoparticles (SNPs) were biofunctionalized using aqueous leaf extracts of G. sylvestre. The anticancer properties of the bioactive compounds and the biofunctionalized SNPs were compared using the HT29 human adenoma colon cancer cell line., Methods: The preliminary phytochemical screening for bioactive compounds from aqueous extracts revealed the presence of alkaloids, triterpenes, flavonoids, steroids, and saponins. Biofunctionalized SNPs were synthesized using silver nitrate and characterized by ultraviolet-visible spectroscopy, scanning electron microscopy, energy-dispersive X-ray analysis, Fourier transform infrared spectroscopy, and X-ray diffraction for size and shape. The characterized biofunctionalized G. sylvestre were tested for its in vitro anticancer activity against HT29 human colon adenocarcinoma cells., Results: The biofunctionlized G. sylvestre SNPs showed the surface plasmon resonance band at 430 nm. The scanning electron microscopy images showed the presence of spherical nanoparticles of various sizes, which were further determined using the Scherrer equation. In vitro cytotoxic activity of the biofunctionalized green-synthesized SNPs (GSNPs) indicated that the sensitivity of HT29 human colon adenocarcinoma cells for cytotoxic drugs is higher than that of Vero cell line for the same cytotoxic agents and also higher than the bioactive compound of the aqueous extract., Conclusion: Our results show that the anticancer properties of the bioactive compounds of G. sylvestre can be enhanced through biofunctionalizing the SNPs using the bioactive compounds present in the plant extract without compromising their medicinal properties.