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The shoots of Asparagus L. are consumed worldwide, although most species belonging to this genus have a restricted range, and several taxa remain unstudied. In this work, a total of four taxa from different locations were scrutinized and compared with cultivated A. officinalis. All shoots were screened for saponins via LC-MS, and in vitro antiproliferative activities against the HT-29 colorectal cancer cell line were assessed via the MTT assay. The total saponins (TS) contained in the crude extracts ranged from 710.0 (A. officinalis) to 1258.6 mg/100 g dw (A. acutifolius). The richness of the compounds detected in this work stands out; a total of 47 saponins have been detected and quantified in the edible parts (shoots) of five taxa of Asparagus. The structure of all the saponins found present skeletons of the furostane and spirostane type. In turn, the structures with a furostane skeleton are divided into unsaturated and dioxygenated types, both in the 20–22 position. The sum of dioscin and derivatives varied largely among the studied taxa, reaching the following percentages of TS: 27.11 (A. officinalis), 18.96 (A. aphyllus), 5.37 (A. acutifolius), and 0.59 (A. albus); while in A. horridus, this compound remains undetected. Aspachiosde A, D, and M varied largely among samples, while a total of seven aspaspirostanosides were characterized in the analyzed species. The hierarchical cluster analysis of the saponin profiles clearly separated the various taxa and demonstrated that the taxonomic position is more important than the place from which the samples were acquired. Thus, saponin profiles have chemotaxonomic significance in Asparagus taxa. The MTT assay showed dose- and time-dependent inhibitory effects of all saponins extracts on HT-29 cancer cells, and the strongest cell growth inhibition was exercised by A. albus and A. acutifolius (GI50 of 125 and 175 µg/mL). This work constitutes a whole approach to evaluating the saponins from the shoots of different Asparagus taxa and provides arguments for using them as functional foods. [ABSTRACT FROM AUTHOR]

The seeds of Boraginaceae species constitute a source of γ-linolenic acid (GLA, 18:3n-6)-rich oils, mainly those of Borago officinalis. However, there are many wild unexplored Boraginaceae taxa that are potentially GLA producers. This work scrutinizes the seeds of several Greek Boraginaceae species for fatty acids (FAs), phenolic compounds, antioxidant activity, and in vitro antiproliferative activity against HT-29 colorectal cancer cells. The FA profiles were checked by GC-FID, and GLA (FA% of total FA) highlights in Symphytum bulbosum (26.2), S. creticum (23.2), and Lithodora zahnii (17.4). The total FA content ranged from 9.2 in Alkanna methanaea to 27.3 g/100 g of seeds in Alkanna corcyrensis. The antioxidant activity of the water/methanol extracts, evaluated by the ABTS and DPPH methods, was in line with other GLA producer seeds. Phenolic acids were analyzed by HPLC-DAD and LC-MS, and rosmarinic acid ranged from 160.2 in S. bulbosum to 479.7 mg/100 g in L. zahnii. The MTT assay showed dose- and time-dependent inhibitory effects of the water/methanol extracts against HT-29 cancer cells, especially those of C. major and L. zahnii (GI50 of 600 and 375 μg/mL for 72 h-exposed cells). This work constitutes the first approach to evaluate the seeds of Boraginaceae taxa from Greece as functional oil providers. [ABSTRACT FROM AUTHOR]

Background: Colorectal cancer (CRC) is among the most common cancers worldwide. In spite of promising improvements, it has a relatively poor prognosis. Therefore, there is a crucial need for discovering novel and efficient therapies. This study aimed to investigate the potential of phycocyanin as a therapeutic agent for CRC, considering its antioxidant and anti-inflammatory properties. Methods: The study used HT-29 and HUVEC cell lines. Cells were exposed to different concentrations of phycocyanin for 24 h. The cytotoxic effects of the phycocyanin on HUVEC and HT-29 cells were evaluated using MTT. Following the treatment, RNA extraction was conducted using TRI reagent. Afterward, real-time PCR (RT-PCR) was performed on Bcl-2, Bax, Akt, PTEN, and ß-actin to study their expression levels. The phycocyanin-treated cells were also stained with Annexin V-FITC to detect any apoptosis and necrosis. Results: Following the MTT assay, the number of living HT-29 cells significantly decreased, with no toxic effect on HUVEC cells. RT-PCR results indicated a significant increase in PTEN and Bax expressions in HT-29 cells due to treatment with phycocyanin, while there was no change in the expression of Bcl-2 and AKT. Flow cytometry on HT-29 cells showed induction of apoptosis and necrosis in 8.79% and 2.9% of cells, respectively, while no necrosis or apoptosis was observed in normal cells. Conclusion: Phycocyanin has a potential for treating colon cancer by affecting AKT/PTEN and Bax/Bcl- 2 and inducing apoptotic genes (Bax and PTEN) while having minimal side effects. Therefore, it could be a promising therapeutic agent for colon cancer. [ABSTRACT FROM AUTHOR]

Colorectal cancer is a frequent malignancy, so its corresponding behaviors are essential to be understood deeply. This work was developed to explore the impacts of Livin on proliferation, migration, invasion, and epithelial-mesenchymal transition of HT-29 colorectal cancer cells. Colorectal cancer cells were extracted from pathological specimens collected from 80 colorectal cancer patients treated at our institution from June 2017 to June 2022. HT-29 human colorectal cancer cells were undertaken as the research materials. Real-time immunofluorescence polymerase chain reaction, Western blot, cell counting kit-8 experiments, cell scratch assays, and Transwell experiments were performed to assess the proliferation, migration, invasion, and epithelial-mesenchymal transition-related markers of the colorectal cancer cells. Livin expression in colorectal cancer cells increased greatly in contrast to that in healthy cells (p<0.05). Compared to normal HT-29, the overexpression of Livin promoted the proliferation, migration, and invasion abilities of colorectal cancer cells, exhibiting great differences (p<0.05). Moreover, levels of matrix metalloproteinase 2, vimentin, slug, and snail in colorectal cancer cells were sharply upregulated (p<0.05), while that of E-cadherin was remarkably downregulated (p<0.05) in contrast to the conditions in normal cells. The findings in this work revealed the crucial role of Livin in HT-29 colorectal cancer cells, suggesting its potential to be a target for colorectal cancer treatment. [ABSTRACT FROM AUTHOR]

Abstract Background Marine cyanobacteria have been known to contain several unique bioactive compounds which have different therapeutic potentials. The current research focuses to identify the efficacy of Nostoc calcicola to counteract the harmful effects of free radicals and testing its anticancer activity against colorectal adenocarcinoma cells (HT-29). Results Methanol is used as a solvent for the extraction of bioactive metabolites from Nostoc calcicola followed by phytochemical screening representing the presence of flavonoids, phenols, tannins, saponins, and steroids to find out bioactive metabolites. Furthermore, evaluation of the extract efficacy revealed the profound ability of Nostoc calcicola to scavenge free radicals by neutralizing different reactive oxygen species. At 100 µg/mL concentration, it inhibited DPPH radicals (73.4%), enhanced phosphomolybdenum reduction (53.5%), displayed ferric-reducing power (55.1%), and finally the extract revealed remarkable hydroxyl radicals scavenging capacity (94.8%), compared to the standards. These compelling results emphasize the robust antioxidant potential of the Nostoc calcicola extract. In vitro, studies demonstrated the selective cytotoxic effects of methanol extracts of Nostoc calcicola on the HT-29 human colorectal cancer cell line, as indicated by IC50 values of 25 µg/mL for the extracts. Treatment with me Nostoc calcicola decreased the cell viability of HT-29 cells followed by consistent morphological changes leading to cytotoxicity. Nuclear condensation and DNA fragmentation were observed using AO/EtBr and DAPI staining. Flow cytometry analysis further confirmed the incidence of apoptosis during the S phase of the cell cycle. Furthermore, western blotting analysis confirmed the activation of caspase 9, a pivotal enzyme in the intrinsic apoptosis pathway, suggesting the ability of Nostoc calcicola to induce apoptosis in HT-29 colorectal cancer cells. Conclusion These findings underscore the potential of Nostoc calcicola as a valuable source of bioactive compounds with antioxidant and anticancer properties, warranting further investigation for their potential therapeutic applications in colorectal cancer. Graphical Abstract

Background: Marine cyanobacteria have been known to contain several unique bioactive compounds which have different therapeutic potentials. The current research focuses to identify the efficacy of Nostoc calcicola to counteract the harmful effects of free radicals and testing its anticancer activity against colorectal adenocarcinoma cells (HT-29). Results: Methanol is used as a solvent for the extraction of bioactive metabolites from Nostoc calcicola followed by phytochemical screening representing the presence of flavonoids, phenols, tannins, saponins, and steroids to find out bioactive metabolites. Furthermore, evaluation of the extract efficacy revealed the profound ability of Nostoc calcicola to scavenge free radicals by neutralizing different reactive oxygen species. At 100 µg/mL concentration, it inhibited DPPH radicals (73.4%), enhanced phosphomolybdenum reduction (53.5%), displayed ferric-reducing power (55.1%), and finally the extract revealed remarkable hydroxyl radicals scavenging capacity (94.8%), compared to the standards. These compelling results emphasize the robust antioxidant potential of the Nostoc calcicola extract. In vitro, studies demonstrated the selective cytotoxic effects of methanol extracts of Nostoc calcicola on the HT-29 human colorectal cancer cell line, as indicated by IC50 values of 25 µg/mL for the extracts. Treatment with me Nostoc calcicola decreased the cell viability of HT-29 cells followed by consistent morphological changes leading to cytotoxicity. Nuclear condensation and DNA fragmentation were observed using AO/EtBr and DAPI staining. Flow cytometry analysis further confirmed the incidence of apoptosis during the S phase of the cell cycle. Furthermore, western blotting analysis confirmed the activation of caspase 9, a pivotal enzyme in the intrinsic apoptosis pathway, suggesting the ability of Nostoc calcicola to induce apoptosis in HT-29 colorectal cancer cells. Conclusion: These findings underscore the potential of Nostoc calcicola as a valuable source of bioactive compounds with antioxidant and anticancer properties, warranting further investigation for their potential therapeutic applications in colorectal cancer. [ABSTRACT FROM AUTHOR]

Isorhamnetin has gained research interest for its anti-inflammatory, anti-proliferative and chemoprotective properties. In this study, human colon adenocarcinoma cells were cultured in the presence or absence of different isorhamnetin concentrations (5–150 μM) for 24 h or 48 h of cultivation to explore the impact on several parameters of viability/proliferation (mitochondrial function using an MTT test, metabolic activity, cell membrane integrity and lysosomal activity using a triple test). The intracellular generation of superoxide radicals using an NBT test and ELISA analysis was performed to observe the biosynthesis of interleukin 8 (IL-8) in cells stimulated with zymosan, as well as in basal conditions. The antiproliferative activity of isorhamnetin was demonstrated by significantly reduced values of mitochondrial and metabolic activity, integrity of cell membranes and lysosomal activity. Its high prooxidant potential was reflected by the significantly elevated generation of superoxides even in cells with low viability status. The anti-inflammatory effect of isorhamnetin was evident due to decreased IL-8 production, and the most significant decline in IL-8 concentration was observed after 24 h treatment in cells with induced inflammation. We demonstrated that isorhamnetin can suppress the proliferation of HT-29 cells, and this effect was correlated with pro-oxidative and anti-inflammatory activity of isorhamnetin. [ABSTRACT FROM AUTHOR]

Colorectal cancer (CRC) is a prevalent malignancy of the digestive tract with the second highest mortality rate globally. Piper nigrum is a widely used traditional medicinal plant, exhibiting antitumor activity against various tumor cells. At present, research on the effect of Piper nigrum on CRC is limited to in vitro cytotoxicity, lacking comprehensive mechanism investigations. This study aimed to explore the inhibitory effect and mechanism of Piper nigrum extract (PNE) on HT-29 cells. Firstly, we identified the chemical components of PNE. Then, MTT assay, colony formation assay, JC-1 staining, and flow cytometry were used to analyze the effect of PNE on HT-29 cells in vitro. A xenograft model, histopathological examination, immunohistochemistry, and western blot were used to evaluate the tumor growth inhibitory activity and mechanism of PNE in vivo. The results indicated that PNE could inhibit cell proliferation and colony formation, reduce mitochondrial membrane potential, induce cell apoptosis in vitro, and inhibit tumor growth in vivo. Furthermore, PNE could regulate p53 and its downstream proteins, and subsequently activate the caspase-3 pathway. In summary, PNE probably induced apoptosis of HT-29 cells through the mitochondrial pathway mediated by p53. All these results suggested that PNE might be a potential natural-origin anti-CRC drug candidate. [ABSTRACT FROM AUTHOR]

Isorhamnetin has gained research interest for its anti-inflammatory, anti-proliferative and chemoprotective properties. In this study, human colon adenocarcinoma cells were cultured in the presence or absence of different isorhamnetin concentrations (5–150 μM) for 24 h or 48 h of cultivation to explore the impact on several parameters of viability/proliferation (mitochondrial function using an MTT test, metabolic activity, cell membrane integrity and lysosomal activity using a triple test). The intracellular generation of superoxide radicals using an NBT test and ELISA analysis was performed to observe the biosynthesis of interleukin 8 (IL-8) in cells stimulated with zymosan, as well as in basal conditions. The antiproliferative activity of isorhamnetin was demonstrated by significantly reduced values of mitochondrial and metabolic activity, integrity of cell membranes and lysosomal activity. Its high prooxidant potential was reflected by the significantly elevated generation of superoxides even in cells with low viability status. The anti-inflammatory effect of isorhamnetin was evident due to decreased IL-8 production, and the most significant decline in IL-8 concentration was observed after 24 h treatment in cells with induced inflammation. We demonstrated that isorhamnetin can suppress the proliferation of HT-29 cells, and this effect was correlated with pro-oxidative and anti-inflammatory activity of isorhamnetin.

Colorectal cancer (CRC) is a prevalent malignancy of the digestive tract with the second highest mortality rate globally. Piper nigrum is a widely used traditional medicinal plant, exhibiting antitumor activity against various tumor cells. At present, research on the effect of Piper nigrum on CRC is limited to in vitro cytotoxicity, lacking comprehensive mechanism investigations. This study aimed to explore the inhibitory effect and mechanism of Piper nigrum extract (PNE) on HT-29 cells. Firstly, we identified the chemical components of PNE. Then, MTT assay, colony formation assay, JC-1 staining, and flow cytometry were used to analyze the effect of PNE on HT-29 cells in vitro. A xenograft model, histopathological examination, immunohistochemistry, and western blot were used to evaluate the tumor growth inhibitory activity and mechanism of PNE in vivo. The results indicated that PNE could inhibit cell proliferation and colony formation, reduce mitochondrial membrane potential, induce cell apoptosis in vitro, and inhibit tumor growth in vivo. Furthermore, PNE could regulate p53 and its downstream proteins, and subsequently activate the caspase-3 pathway. In summary, PNE probably induced apoptosis of HT-29 cells through the mitochondrial pathway mediated by p53. All these results suggested that PNE might be a potential natural-origin anti-CRC drug candidate.

Purpose: Radiotherapy is one of the most important therapeutic options used to treat cancers. Radiation effects can be improved using nanoparticles and chemotherapeutic drugs as radiosensitizing agents. The aim of the present study was to evaluate the effects of folic acidconjugated gold nanoparticles (GNP-F) in combination with doxorubicin (DOX) and x-Ray irradiation in colorectal cancer (CRC) cell line (HT-29). Methods: The cell viability assay (WST-1) was performed to study the cytotoxic effects of different concentrations of DOX and GNP-F after 24 and 48 hours treatments. Then, the effects of the GNP-F, X-Ray irradiation, and DOX drug in single and combined treatments were examined after 24 and 48 hours treatment with effective doses. Likewise, the caspase 3 gene expression ratio and the caspase 3 activity were assessed after 48 h treatment. Moreover, the malondialdehyde (MDA) level was determined in treated and untreated cells. Results: When GNP-F (at a concentration of 70 μM) was combined with X-ray irradiation (2 Gy) and DOX drug, induced more cytotoxic effects compared to the control group. The results of cell viability assay showed that GNP-F + X-Ray in combination with a low concentration of DOX (0.25 x IC50) enhanced the cytotoxic effects of cells compared to related single treatments. Caspase 3 gene expression ratio and caspase 3 activity increased in double and triple combination treatments in comparison with the single groups. Moreover, the MDA level increased in triple combination compared to the single treatments. Conclusion: Our findings confirmed the potential anti-cancer effects of the GNP-F and DOX in combination with X-Ray irradiation in CRC cells. [ABSTRACT FROM AUTHOR]

Colorectal cancer is the third most malignant cancer occurring around the world. Effective prevention and treatment have been increasingly the focus of global attention. Long‐term diet of fermented dairy inhibits proliferation of colon cancer cell, which is considered that not only live lactic acid bacteria but also the secreted exopolysaccharides exert the function. In this scenario, this study aimed to investigate the mechanism of growth inhibition on HT‐29 cells induced in vitro by exopolysaccharides isolated from Lactobacillus paracasei subsp. paracasei M5L (M5‐EPSs). HT‐29 cells which were treated by a set of concentrations of M5‐EPSs have been investigated of cell viability, characteristic changes, cell cycle distribution, and redox system. The results demonstrated that M5‐EPSs treatments induced HT‐29 cell apoptosis and resulted in upregulation of ROS levels and downregulation of antioxidant enzyme activities, leading to an imbalance in the oxidation system in HT‐29 cells. In response to M5‐EPSs, endogenous ER stress (ERS) markers, including GRP78, ATF4, and CHOP, were transcriptionally altered so that activating the ERS in HT‐29 cells. After NAC treatment, the oxidative stress was inhibited, and the expression of GRP78 and CHOP was significantly decreased, indicating that oxidative stress can significantly affect the ERS pathway. Furthermore, it suggested that the occurrence of apoptosis was associated with Bcl‐2 gene family. In conclusion, this study demonstrated that M5‐EPSs can induce HT‐29 cells apoptosis by destroying the redox system through activation of the ERS signaling pathway. [ABSTRACT FROM AUTHOR]

This research sought to assess the anticancer effects of Lactobacillus acidophilus CICC 6074 and its underlying mechanism of action. L. acidophilus CICC 6074 inhibited HT-29 cell activity and induced its apoptosis in a dose- and time-dependent manner. HT-29 cells exposed to L. acidophilus CICC 6074 showed mitochondrial membrane potential (ΔΨm) loss, as well as Cyt c is released from the mitochondria into the cytoplasm, causing Bax, Caspase-9 and Caspase-3 to be up-regulated and Bcl-2 to be down-regulated. To further explore the mechanism by which L. acidophilus CICC 6074 achieves its anti-cancer effect, a HT-29 cell-induced colon cancer mouse model was used. Compared with a colon cancer model control group, the high-dose L. acidophilus CICC 6074 group showed obvious colon cancer cell apoptosis as measured by histological morphology, electron microscopy, and terminal dexynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) assay. An in vivo study showed that L. acidophilus CICC 6074 induced colon cancer apoptosis by up-regulating Bax, down-regulating Bcl-2, releasing Cyt c from the mitochondria into the cytoplasm, and activating Caspase-3 and Caspase-9. These results demonstrate that L. acidophilus CICC 6074 exerts anticancer effects via the activation of mitochondrial pathways which then induce apoptosis. The research could provide new ideas and theoretical basis for development of probiotics functional food market.

α-Linolenic acid (ALA, 18:3n-3) and γ -gamma linolenic acid (GLA, 18:3n-6) are polyunsaturated fatty acids (PUFA) that improve the human health. The present study focused on testing the in vitro antitumor actions of pure ALA and GLA on the HT-29 human colorectal cancer cell line. Cell viability was checked by MTT ((3-(4,5-Dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide) test, cell membrane damage by the lactate dehydrogenase assay, apoptosis was tested by both caspase-3 activity trial and transmission electron microscopy images, and protein composition was analyzed by quantitative proteomics analysis. MTT test revealed IC50 values of 230 and 255 μM for ALA and GLA, respectively, at 72 h. After 24 h of incubation, both ALA and GLA induced apoptosis on HT-29 colorectal cancer cells according to the caspase-3 assay and microscopy images. SWATH/MS analysis evidenced that ALA significantly affected the mitochondrial protein import pathway and the citric acid cycle pathway, while GLA did not significantly affect any particular pathway. In summary, both ALA and GLA showed concentration-dependent inhibitory effects on HT-29 cells viability and induced cell death by apoptosis. ALA significantly affected cellular pathways, while GLA does not have specific actions on either pathway. Both n-3 and n-6 C18 PUFA are bioactive food components useful in the colorectal cancer prevention. [ABSTRACT FROM AUTHOR]

Aim: The aim of this investigation was to evaluate the potential of folic acid-tailored solid lipid nanoparticles (SLNs) for encapsulation as well as for in vitro cytotoxicity study of irinotecan hydrochloride trihydrate (IHT) against colorectal cancer (CRC) by using HT-29 cells. Methods: Solvent diffusion technique was employed for the preparation of SLNs. Further, the formulations were optimised via three-level, three-factor Box–Behnken design (BBD). Results: The uncoupled SLNs (IRSLNs) and folic acid-coupled SLNs (IRSLNFs) formulations revealed not only high %entrapment efficiency but also small particle size. Moreover, in vitro drug release results from IRSLNs and IRSLNFs confirmed that they followed sustained-release effect for up to 144 h. Whereas, in vitro cell viability study against HT-29 cell line suggested significantly (p < 0.05) higher cytotoxicity (IC50 = 15 µg/ml) of IRSLNFs over IRSLNs and IHT solution. Conclusions: Outcomes suggested that the engineered IRSLNFs hold great potential for targeting CRC for an extended period of time. [ABSTRACT FROM AUTHOR]

Efficient microwave-assisted chemical processes were applied to the synthesis of an array of novel N-(4-methoxyphenylamino)-2-methyl benzo-, pyrido- or pyrazino-thieno[3,2-d]pyrimidin-4-amine derivatives. These heteroaromatic systems were envisioned as potent bioisosteric analogues of MPC-6827, an anticancer agent previously developed until phase II clinical studies. A brief evaluation and comparison of their antiproliferative activity on HT-29 and Caco-2, two human colorectal cancer cell lines, were also reported. At the tested concentrations (5 and 10 µM), thieno[3,2-d]pyrimidin-4-amines 4a and 4c exhibited an inhibitory effect similar to MPC-6827 on human colorectal cancer cell proliferation.

The research for innovative treatments against colon adenocarcinomas is still a great challenge. Acacia catechu Willd. heartwood extract (AC) has health-promoting qualities, especially at the gastrointestinal level. This study characterized AC for its catechins content and investigated the apoptosis-enhancing effect in human colorectal adenocarcinoma HT-29 cells, along with its ability to spare healthy tissue. MTT assay was used to describe the time course, concentration dependence and reversibility of AC-mediated cytotoxicity. Cell cycle analysis and AV-PI and DAPI-staining were performed to evaluate apoptosis, together with ROS formation, mitochondrial membrane potential (MMP) changes and caspase activities. Rat ileum and colon rings were tested for their viability and functionality to explore AC effects on healthy tissue. Quantitative analysis highlighted that AC was rich in (±)-catechin (31.5 ± 0.82 mg/g) and (−)-epicatechin (12.5 ± 0.42 mg/g). AC irreversibly decreased cell viability in a concentration-dependent, but not time-dependent fashion. Cytotoxicity was accompanied by increases in apoptotic cells and ROS, a reduction in MMP and increases in caspase-9 and 3 activities. AC did not affect rat ileum and colon rings’ viability and functionality, suggesting a safe profile toward healthy tissue. The present findings outline the potential of AC for colon cancer treatment.

Colorectal cancer is the third most common malignancy in the world having a high mortality rate. Flavonoids possess many biological activities including anti-cancer activity. lawsonaringenin (LSG) is a flavonoid isolated from leaves of Lawsonia alba Lam. The objective of this study was to demonstrate the anti-cancer potential of LSG in colorectal cancer for the first time. The HT-29 cells were treated with LSG or 5-fluoruracil, as a positive control, to determine its effect on cell cytotoxicity by a MTT cell proliferation assay, and cell cycle progression and apoptosis using flowcytometry. We also determined the mechanisms underlying LSG-mediated growth inhibition of HT-29 cells by by investigating the expression of key oncogenes and apoptosis genes using q-RT PCR and immunocytochemical analysis. The cell cytotoxicity data showed that the IC50 value of LSG was significantly less than the IC50 value of 5-FU (50 µM). The anti-proliferative effect of LSG was mediated by arresting cells in the S phase of the cell cycle which then led to the induction of apoptosis the q-RT PCR and immunocytochemical analysis showed that LSG reduced the expression of ß-catenin (non-phosphorylated) and its downstream signalling target c-Myc, whereas it increased the phosphorylation of ß-catenin. Furthermore, LSG also downregulated the expression of oncogene K-Ras and anti-apoptotic proteins, Bcl-2, and Bcl-xL. In conclusion, our data demonstrates that LSG exerted its anti-tumor activity by arresting the cell cycle in S phase, and by downregulating the expression of oncogenes including ß-catenin, c-Myc, K-Ras and anti-apoptosis proteins Bcl-2 and Bcl-xL. This study suggests a potential use of natural flavonoid, lawsonaringenin, to attenuate colorectal cancer growth; however, further pre-clinical/clinical studies are required to establish its role as a therapeutic agent. [ABSTRACT FROM AUTHOR]

• Isolation and characterization of nine sappinane-4-one type homoisoflavonoids from the bulbs of Scilla bisotunensis Speta. • In vitro cytotoxic effects of the homoisoflavonoids against HT-29 colon cancer cells. • Determination of absolute configuration of sappinane-4-one homoisoflavonoids by circular dichroism experiment. • A salient binding affinity of bioactive homoisoflavonoids, 8 and 9 , with the Src kinase domain enzyme. • Homoisoflavonoids 1 ‒ 3 and 5 are suggested as chemotaxonomic markers for S. bisotunensis. Phytochemical investigation of the chloroform extract of Scilla bisotunensis Speta. bulbs led to the isolation and structure elucidation of six sappinane-4-one (1-4, 7, 9) and three 3,9-dehydrosappinan-4-one (5, 6, 8) homoisoflavonoids. The structures of compounds 1 - 9 were established based on extensive NMR, ESIMS, UV, and ECD spectra, and also by comparison with the previously reported spectroscopic data. The in vitro cytotoxicity of sappanin-type homoisoflavonoids 1 - 9 was assayed against HT-29 human colorectal cancer cells using sulforhodamine B assay. Compounds 8 and 9 showed higher cytotoxic effects with IC 50 of 5.3 and 6.1 µ g/mL, respectively, while compounds 3, 5 , and 7 exhibited moderate activity with IC 50 ranging from 25 to 37 µ g/mL. Interestingly, molecular docking studies revealed that compounds 1‒9 could be potential sarcoma kinase domain inhibitors with binding energies ranging from ‒ 7.37 (6) to ‒8.34 (9) kcal/mol when compared with the native inhibitor, purvalanol A of ‒ 7.90 kcal/mol. All homoisoflavonoids, except 1 and 5 are reported for the first time in a member of the Scilla genus, while, to the best of our knowledge this is the first phytochemical and biological study on S. bisotunensis. The plant exhibited good chemotaxonomic relationships to the genus Eucomis L'Hér. on the basis of comparing their chemical constituents with those reported in the related plant families. [Display omitted] [ABSTRACT FROM AUTHOR]

[Display omitted] • Seeds of several Buglossoides taxa were surveyed for fatty acids (FA) and phenolics. • Stearidonic acid reached high amounts in wild B. arvensis samples. • Rosmarinic and lithospermic acids were the predominant phenolics. • Unsaturated FA allow grouping Buglossoides taxa as botanical criteria do. • The phenolics of B. arvensis were active against HT-29 colorectal cancer cells. Ahiflower oil© is extracted from the seeds of Buglossoides arvensis , which contains high amounts of stearidonic acid (SDA, 18:4 n -3), while its phenolic composition still is unreported. Moreover, several Buglossoides taxa remain unstudied and could become natural sources of SDA. In this work, seeds of several Buglossoides taxa and Ahiflower oil© were screened for fatty acids, phenolic compounds, and in vitro antiproliferative activities against colorectal cancer cells. Four flavonoids and 16 phenolic acids were identified and quantified. Among Buglossoides taxa, the highest amounts of phenolic compounds were found in samples collected in Spain, under a warm Mediterranean climate. Rosmarinic and lithospermic acids were the main phenols found in Buglossoides seeds. The MTT assay showed dose- and time-dependent inhibitory effects of B. arvensis extracts on HT-29 cancer cells, with a GI 50 value of ∼280 μg/mL after 72 h of cell exposure to seed extracts. The latter showed lower antiproliferative activity than that of pure phenolics due to the simultaneous presence of other compounds in the extracts, as evidenced by 1H NMR. This work constitutes the first approach to evaluate the seeds of several Buglossoides taxa as functional oils-providers to use them as functional foods. [ABSTRACT FROM AUTHOR]

The Ca2+-activated Cl− channel ANO1 is widely expressed in epithelial cells, and ANO1 upregulation is implicated in the oncogenesis of many epithelium-originated cancers. However, whether ANO1 plays a causal role in the tumorigenesis of colorectal cancer remains largely unknown. Here, we show that ANO1 channel protein is upregulated in human colorectal cancer tissue samples and its upregulation is correlated with the TNM staging, histological type, pathological differentiation and poor prognosis. Knockdown or pharmacological inhibition of ANO1 suppresses colorectal cancer cell proliferation and induces cell apoptosis. Furthermore, ANO1 knockdown inhibits the growth of subcutaneous xenograft tumors implanted with colorectal cancer HT-29 cells in nude mice. Mechanically, knockdown of endogenous ANO1 inactivates the Wnt/β-catenin signaling through downregulating critical components, such as Frizzled protein 1, β-catenin and upregulating GSK3β. Taken together, our results demonstrate that ANO1 upregulation is involved in the tumorigenesis of colorectal cancer, and inhibition of ANO1 upregulation or inactivating downstream Wnt/β-catenin signaling may have therapeutic potential for colorectal cancer. • TMEM16A/ANO1 channel protein is upregulated in human tissue samples of colorectal cancer (CRC). • TMEM16A/ANO1 protein upregulation is correlated with the clinical TNM staging, and poor survival rate of CRC patients. • Inhibition of TMEM16A/ANO1 suppresses CRC cell proliferation, xenograft tumor growth and induces cell apoptosis. • TMEM16A/ANO1 downregulation suppresses tumor growth through inactivating the Wnt/β-catenin signaling. [ABSTRACT FROM AUTHOR]

Objective: Neurotrophin and neuropeptide pathways are emerging targets in cancer. Here we show that brain-derived neurotrophic factor (BDNF) and its receptor, TrkB, are present in colorectal cancer and that BDNF levels are increased in tumors compared to nontumor tissue. In addition, we investigate the role of BDNF in influencing the response of colorectal cancer cells to inhibition of gastrin-releasing peptide receptors (GRPR). Methods: Fresh-frozen sporadic colorectal adenocarcinoma specimens and adjacent nonneoplastic tissue from 30 patients, as well as paraffin-embedded colorectal cancer samples from 21 patients, were used in this study. Cell proliferation and mRNA and protein levels were examined in HT-29 or SW620 cells treated with a GRPR antagonist, human recombinant BDNF (hrBDNF), a Trk antagonist K252a, or cetuximab. Results: Expression of BDNF and TrkB was detected in tumor samples and cell lines. BDNF levels were higher in tumor samples compared to nonneoplastic tissue. BDNF expression and secretion were increased by GRPR blockade in HT-29 cells through a mechanism dependent on epidermal growth factor receptors. Treatment with hrBDNF prevented the effect of GRPR blockade on cell proliferation, whereas a Trk inhibitor reduced proliferation. Conclusions: BDNF and TrkB are present in colorectal cancer and might contribute to resistance to GRPR antagonists. Copyright © 2011 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Calystegia soldanella is a halophyte and a perennial herb that grows on coastal sand dunes worldwide. Extracts from this plant have been previously revealed to have a variety of bioactive properties in humans. However, their effects on colorectal cancer cells remain poorly understood. In the present study, the potential biological activity of C. soldanella extracts in the colorectal cancer cell line HT-29 was examined. First, five solvent fractions [n-hexane, dichloromethane (DCM), ethyl acetate, n-butanol and water] were obtained from the crude extracts of C. soldanella through an organic solvent extraction method. In particular, the DCM fraction was demonstrated to exert marked dose- and time-dependent inhibitory effects according to results from the cell viability assay. Data obtained from the apoptosis assay suggested that the inhibition of HT-29 cell viability induced by DCM treatment was attributed to increased apoptosis. The apoptotic rate was markedly increased in a dose-dependent manner, which was associated with the protein expression levels of apoptosis-related proteins, including increased Fas, Bad and Bax, and decreased pro-caspase-8, Bcl-2, Bcl-xL, pro-caspase-9, pro-caspase-7 and pro-caspase-3. A mitochondrial membrane potential assay demonstrated that more cells became depolarized and the extent of cytochrome c release was markedly increased in a dose-dependent manner in HT-29 cells treated with DCM. In addition, cell cycle analysis confirmed S-phase arrest following DCM fraction treatment, which was associated with decreased protein expression levels of cell cycle-related proteins, such as cyclin A, CDK2, cell division cycle 25 A and cyclin dependent kinase inhibitor 1. Based on these results, the present study suggested that the DCM fraction of the C. soldanella extract can inhibit HT-29 cell viability whilst inducing apoptosis through mitochondrial membrane potential regulation and S-phase arrest. These results also suggested that the DCM fraction has potential anticancer activity in HT-29 colorectal cells. Further research on the composition of the DCM fraction is warranted. [ABSTRACT FROM AUTHOR]

Abstract: Colorectal cancer is the most abundant cause of cancer mortality in the Western world. Nutrition and the microbial flora are considered to have a marked influence on the risk of colorectal cancer, the formation of butyrate and other short-chain fatty acids (SCFAs) possibly playing a major role as chemopreventive products of microbial fermentation in the colon. In this study, we investigated the effects of butyrate, other SCFAs, and of a number of phenolic SCFA and trans-cinnamic acid derivatives formed during the intestinal degradation of polyphenolic constituents of fruits and vegetables on global histone deacetylase (HDAC) activity in nuclear extracts from colon carcinoma cell cultures using tert-butoxycarbonyl-lysine (acetylated)-4-amino-7-methylcoumarin (Boc-Lys(Ac)-AMC) as substrate. Inhibition of HDAC activity, e.g., by butyrate, is related to a suppression of malignant transformation and a stimulation of apoptosis of precancerous colonic cells. In nuclear extracts from HT-29 human colon carcinoma cells, butyrate was found to be the most potent HDAC inhibitor (IC50=0.09 mM), while other SCFAs such as propionate were less potent. In the same assay, p-coumaric acid (IC50=0.19 mM), 3-(4-OH-phenyl)-propionate (IC50=0.62 mM) and caffeic acid (IC50=0.85 mM) were the most potent HDAC inhibitors among the polyphenol metabolites tested. Interestingly, butyrate was also the most potent HDAC inhibitor in a whole-cell HeLa Mad 38-based reporter gene assay, while all polyphenol metabolites and all other SCFAs tested were much less potent; some were completely inactive. The findings suggest that butyrate plays an outstanding role as endogenous HDAC inhibitor in the colon, and that other SCFAs and HDAC-inhibitory polyphenol metabolites present in the colon seem to play a much smaller role, probably because of their limited levels, their marked cytotoxicity and/or their limited intracellular availability. [Copyright &y& Elsevier]

Human colorectal cancer (CRC) is treated with 5-fluorouracil (5-Fu). However, this drug causes various adverse effects, suggesting the benefit of developing safer natural-based drugs with high specificity and tolerable adverse effects. Dendrobium officinale has been used as a functional food and important medicine in China for more than a thousand years. The natural polysaccharides obtained from D. officinale (DOPW-1) show inhibitory activity against cancer cells. This study aimed to comparatively investigate the anticancer activity of DOPW-1 and mannose (Mann) on HT-29 cells and explored the possible underlying molecular mechanisms of CRC treatment. HT-29 cells were treated with 5-Fu, DOPW-1, and Mann or their combinations. Cell viability and apoptosis were examined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and Hoechst 33342 with annexin V-fluorescein isothiocyanate/propidium iodide assays, respectively. DOPW-1 showed good antitumor effects on HT-29 cells and significantly potentiated the anticancer activities of 5-Fu in vitro and in vivo. DOPW-1 significantly inhibited tumor metastasis in the zebrafish model. Furthermore, the monosaccharide composition of DOPW-1 might not have an important effect on its antitumor activity. The results of Western blot analysis suggested that DOPW-1 alone and in combination with 5-Fu induced cell apoptosis using the mitochondria-dependent intrinsic apoptotic pathway. Thus, DOPW-1 is a potential source of natural polysaccharides for CRC therapy alone or in combination with 5-Fu. [Display omitted] • Dendrobium officinale polysaccharide (DOPW-1) showed good antitumor effects. • DOPW-1 significantly potentiated anticancer effects of 5-Fu in vitro and in vivo. • DOPW-1 significantly inhibited tumour metastasis in the zebrafish model. • Mannose content of DOPW-1 did not mediate antitumor activity against CRC. • DOPW-1 induced apoptosis in CRC using the mitochondria-dependent apoptotic pathway. [ABSTRACT FROM AUTHOR]

• The first study of the anti-cancer mechanism of Lactobacillus acidophilus CICC 6074 was carried out. • L. acidophilus CICC 6074 exerts anticancer effects in HT-29 cell-induced colon cancer mouse model. • L. acidophilus CICC 6074 may induce HT-29 cell apoptosis through the mitochondrial pathway. • The research may provide theoretical basis for development of probiotics functional food market. This research sought to assess the anticancer effects of Lactobacillus acidophilus CICC 6074 and its underlying mechanism of action. L. acidophilus CICC 6074 inhibited HT-29 cell activity and induced its apoptosis in a dose- and time-dependent manner. HT-29 cells exposed to L. acidophilus CICC 6074 showed mitochondrial membrane potential (ΔΨm) loss, as well as Cyt c is released from the mitochondria into the cytoplasm, causing Bax, Caspase-9 and Caspase-3 to be up-regulated and Bcl-2 to be down-regulated. To further explore the mechanism by which L. acidophilus CICC 6074 achieves its anti-cancer effect, a HT-29 cell-induced colon cancer mouse model was used. Compared with a colon cancer model control group, the high-dose L. acidophilus CICC 6074 group showed obvious colon cancer cell apoptosis as measured by histological morphology, electron microscopy, and terminal dexynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) assay. An in vivo study showed that L. acidophilus CICC 6074 induced colon cancer apoptosis by up-regulating Bax, down-regulating Bcl-2, releasing Cyt c from the mitochondria into the cytoplasm, and activating Caspase-3 and Caspase-9. These results demonstrate that L. acidophilus CICC 6074 exerts anticancer effects via the activation of mitochondrial pathways which then induce apoptosis. The research could provide new ideas and theoretical basis for development of probiotics functional food market. [ABSTRACT FROM AUTHOR]

Efficient microwave-assisted chemical processes were applied to the synthesis of an array of novel N-(4-methoxyphenylamino)-2-methyl benzo-, pyrido- or pyrazino-thieno[3,2-d]pyrimidin-4-amine derivatives. These heteroaromatic systems were envisioned as potent bioisosteric analogues of MPC-6827, an anticancer agent previously developed until phase II clinical studies. A brief evaluation and comparison of their antiproliferative activity on HT-29 and Caco-2, two human colorectal cancer cell lines, were also reported. At the tested concentrations (5 and 10 µM), thieno[3,2-d]pyrimidin-4-amines 4a and 4c exhibited an inhibitory effect similar to MPC-6827 on human colorectal cancer cell proliferation. [ABSTRACT FROM AUTHOR]

The research for innovative treatments against colon adenocarcinomas is still a great challenge. Acacia catechu Willd. heartwood extract (AC) has health-promoting qualities, especially at the gastrointestinal level. This study characterized AC for its catechins content and investigated the apoptosis-enhancing effect in human colorectal adenocarcinoma HT-29 cells, along with its ability to spare healthy tissue. MTT assay was used to describe the time course, concentration dependence and reversibility of AC-mediated cytotoxicity. Cell cycle analysis and AV-PI and DAPI-staining were performed to evaluate apoptosis, together with ROS formation, mitochondrial membrane potential (MMP) changes and caspase activities. Rat ileum and colon rings were tested for their viability and functionality to explore AC effects on healthy tissue. Quantitative analysis highlighted that AC was rich in (±)-catechin (31.5 ± 0.82 mg/g) and (−)-epicatechin (12.5 ± 0.42 mg/g). AC irreversibly decreased cell viability in a concentration-dependent, but not time-dependent fashion. Cytotoxicity was accompanied by increases in apoptotic cells and ROS, a reduction in MMP and increases in caspase-9 and 3 activities. AC did not affect rat ileum and colon rings' viability and functionality, suggesting a safe profile toward healthy tissue. The present findings outline the potential of AC for colon cancer treatment. [ABSTRACT FROM AUTHOR]

• Anti-proliferative and apoptosis effects of a benzochromene derivative (4-FC) was analyzed against HT-29 cell line. • The QRT-PCR was used to analysis the effects of 4-FC on mRNA expression levels of Bcl-2, Bax and caspase-3, -8 and -9. • The binding features of 4-FC with ctDNA have been investigated. • The compound possesses significant cytotoxic and apoptosis inducing properties. 3-Amino-1-aryl-1H-benzo f chromene-2-carbonitrile derivatives were synthesized from three-component reaction of arylaldehyde, malononitrile and 2-naphthol in the presence of 1, 4-bis(4-ferrocenylbutyl)piperazine as a new catalyst. Cytotoxic potencies of the compounds were tested on HT-29 cells. 3-Amino-1-(4-fluorophenyl)-1 H -benzo f chromene-2-carbonitrile (4c) was more active among these compounds and was selected for further studies. Apoptosis was investigated by acridine orange/ethidium bromide (AO/EtBr) double staining and flow cytometry. The qRT-PCR was used to analyze the expression of pro- and anti-apoptotic genes. The binding attributes of 4c with calf thymus DNA (ctDNA) was examined using multi-spectroscopic measurements. We found that 4c had potent cytotoxic activity against HT-29 cells with an IC 50 value of 60 µM through induction of cell cycle arrest in the sub-G1 phase and apoptosis. RT-PCR analysis demonstrated down-regulation of Bcl-2 expression, while the expression of Bax, caspase-3, -8 and -9 genes was up-regulated in HT-29 cells incubated with 4c compared with control cells. These studies revealed that 4c interacts with DNA through groove binding mode with the intrinsic binding constant (K b) of 3 × 102 M−1. Thus, 4c is a valuable candidate for further evaluation as a new series of potent chemotherapeutic family in colon cancer treatment. [ABSTRACT FROM AUTHOR]

Colorectal cancer (CRC) is one of the most common and mortal types of cancer. There is increasing evidence that some polyunsaturated fatty acids (PUFAs) exercise specific inhibitory actions on cancer cells through different mechanisms, as a previous study on CRC cells demonstrated for two very long-chain PUFA. These were docosahexaenoic acid (DHA, 22:6n3) and arachidonic acid (ARA, 20:4n6) in the free fatty acid (FFA) form. In this work, similar design and technology have been used to investigate the actions of both DHA and ARA as monoacylglycerol (MAG) molecules, and results have been compared with those obtained using the corresponding FFA. Cell assays revealed that ARA- and DHA-MAG exercised dose- and time-dependent antiproliferative actions, with DHA-MAG acting on cancer cells more efficiently than ARA-MAG. Sequential window acquisition of all theoretical mass spectra (SWATH)—mass spectrometry massive quantitative proteomics, validated by parallel reaction monitoring and followed by pathway analysis, revealed that DHA-MAG had a massive effect in the proteasome complex, while the ARA-MAG main effect was related to DNA replication. Prostaglandin synthesis also resulted as inhibited by DHA-MAG. Results clearly demonstrated the ability of both ARA- and DHA-MAG to induce cell death in colon cancer cells, which suggests a direct relationship between chemical structure and antitumoral actions. [ABSTRACT FROM AUTHOR]

Two varieties of common beans (Phaseolus vulgaris L.), Bayo Victoria and Negro 8025, were evaluated to determine the effect on cellular viability and mechanisms involved in apoptosis pathways, using a cellular model with HT‐29 cells. Aqueous methanolic (50:50) extracts from cooked beans were analyzed for phenolic composition, identifying greater diversity of phenolic compounds in Bayo Victoria extracts. However, Negro 8025 showed greater phenolic content and cytotoxicity effects at lower media inhibitory concentrations, and greater effectiveness to activate apoptotic pathways. Proteins related to the arrest of cell cycle were modulated by both bean cultivars. Qualitative analysis by HPLC‐PAD and HPLC‐MS systems of phenolic compounds in common bean extracts showed mainly hydroxybenzoic and hydroxycinnamic acids, flavonols, and monomeric flavan‐3‐ols. Bioactive phenolics such as catechin, kaempferol, and ferulic acid were found in both cultivars as well anticancer phytochemicals such as quercetin, protocatechuic acid, myricetin, naringenin and their derivatives, and procyanidins. Practical applications: Polyphenols in common beans (Phaseolus vulgaris L.) cultivars processed by canning display chemoprotective potential as they activate mechanisms involved in apoptosis pathways. Phenolics in common beans modulate 28 proteins related to apoptotic processes. Therefore, a diet including canned beans (particularly darker varieties) might represent health benefits and cancer‐preventive effects. [ABSTRACT FROM AUTHOR]