Colorectal cancer (CRC) is the second greatest cause of cancer-related death in the world and chemotherapy, as an important part of CRC treatment, has some drawbacks, including systemic toxicity. Therefore, it is crucial to discover new and more effective CRC treatment plans. Rheum khorasanicum (R. khorasanicum) is a medicinal plant with high flavonoids, stilbenes, and anthraquinone contents, so it can be a potential source of antioxidants and can be used for therapeutic purposes and trigger apoptosis in cancer cells. In this study, we investigated the effects of hydroalcoholic root extract of R. khorasanicum treatment on inducing mitochondrial apoptosis of HT-29 and Caco-2 human colorectal adenocarcinoma cells. Firstly, the total phenolic and flavonoid content was determined. Then, the cytotoxic effects of R. khorasanicum on cells of three different types, including HT-29 and Caco-2 colon cancer cells as well as normal 3T3 cells were assessed using the MTT assay. To investigate the characteristics of cellular death, flow cytometry, and western blotting were performed. The results of this study indicated considerable phenolic (356.4±9.4 GAE/gDW) and flavonoid (934.55±17.1 QE/gDW) contents in R. khorasanicum. MTT assay\'s finding indicated that 100, 60, and 30μg/mL concentrations of R. khorasanicum reduce cell viability in HT-29 and Caco-2 cell lines significantly (P<0.05). It has been also revealed that R. khorasanicum extract induces apoptosis rather than necrosis in these cell lines. Moreover, Bcl-2 expression was significantly reduced in both HT-29 and Caco-2 cell lines, while Bax and cleaved caspase-3 expression soared considerably in the groups under R. khorasanicum treatment (P<0.05). In conclusion, our findings have suggested that high phenol and flavonoid contents of R. khorasanicum root extract possibly play an important role in cell cytotoxicity and apoptosis induction in HT-29 and Caco-2 colon cancer cells.

BACKGROUND: Since cancer is one of the most prevalent diseases in the world, further studies are needed to identify the effective therapeutic modalities. The second deadliest and third most common cancer is colorectal cancer (CRC). Papaya (Carica papaya Linn) seeds offer anti-cancer properties that can cure various types of cancer, such as liver and prostate cancer.
METHODS: The study aimed to evaluate the anti-cancer activity of Carica papaya seed extract on colorectal cancer cell lines (Caco-2) and used techniques to assess the anti-cancer potential. The effectiveness of SE on cell proliferation and the viability of HTB-37 Caco-2 and C-166 cells were assessed using the MTT test. Real-time RT-PCR was used to measure gene expression levels and evaluate the activity of genes involved in apoptosis, including caspase-3, p53, Cycs, and Bcl-2. Finally, flow cytometry was used to analyze apoptosis induction by detecting changes in cell morphology and DNA content.
RESULTS: The study showed that the MTT reduction assay was dependent on cancer cell type and concentration of SE compared to the control cells and C-166, with a mean IC50 value of 9.734 ug/ml. The cytotoxicity was accompanied by some morphological alterations in the colorectal cancer cell line (Caco-2). The expression of the genes for p53, Cycs, and caspase-3 was substantially up-regulated, while Bcl-2 was dramatically down-regulated compared to control cells. The cell cycle arrested at the G2-M phase and the presence of early and late apoptotic characteristics post-treatment increased the apoptotic profile.
CONCLUSIONS: It concluded that papaya seeds aqueous extract could act as a novel therapeutic option for colorectal cancer (CRC).

A key component of efforts to identify the biological and drug-specific aspects contributing to therapeutic failure or unexpected exposure-associated toxicity is the study of drug-intestinal barrier interactions. While methods supporting such assessments are widely described for human therapeutics, relatively little information is available for similar evaluations in support of veterinary pharmaceuticals. There is, therefore, a critical need to develop novel approaches for evaluating drug-gut interactions in veterinary medicine. Three-dimensional (3D) organoids can address these difficulties in a reasonably affordable system that circumvents the need for more invasive in vivo assays in live animals. However, a first step in developing such systems is understanding organoid interactions in a 2D monolayer. Given the importance of orally administered medications for meeting the therapeutic need of companion animals, we demonstrate growth conditions under which canine-colonoid-derived intestinal epithelial cells survive, mature, and differentiate into confluent cell systems with high monolayer integrity. We further examine the applicability of this canine-colonoid-derived 2D model to assess the permeability of three structurally diverse, passively absorbed β-blockers (e.g., propranolol, metoprolol, and atenolol). Both the absorptive and secretive apparent permeability (Papp) of these drugs at two different pH conditions were evaluated in canine-colonoid-derived monolayers and compared with that of Caco-2 cells. This proof-of-concept study provides promising preliminary results with regard to the utility of canine-derived organoid monolayers for species-specific assessments of therapeutic drug passive permeability.

trans-Resveratrol can be catabolized by the gut microbiota to dihydroresveratrol, 3,4\'-dihydroxy-trans-stilbene, lunularin, and 4-hydroxydibenzyl. These metabolites can reach relevant concentrations in the colon. However, not all individuals metabolize RSV equally, as it depends on their RSV gut microbiota metabotype (i.e., lunularin producers vs. non-producers). However, how this microbial metabolism affects the cancer chemopreventive activity of stilbenes and their microbial metabolites is poorly known. We investigated the structure-antiproliferative activity relationship of dietary stilbenes, their gut microbial metabolites, and various analogs in human cancer (Caco-2 and HT-29) and non-tumorigenic (CCD18-Co) colon cells. The antiproliferative IC50 values of pterostilbene, oxy-resveratrol, piceatannol, resveratrol, dihydroresveratrol, lunularin, 3,4\'-dihydroxy-trans-stilbene, pinosylvin, dihydropinosylvin, 4-hydroxy-trans-stilbene, 4-hydroxydibenzyl, 3-hydroxydibenzyl, and 4-trans-stilbenemethanol were calculated. IC50 values were correlated with 34 molecular characteristics by bi- and multivariate analysis. Little or no activity on CCD18-Co was observed, while Caco-2 was more sensitive than HT-29, which was explained by their different capacities to metabolize the compounds. Caco-2 IC50 values ranged from 11.4 ± 10.1 μM (4-hydroxy-trans-stilbene) to 73.9 ± 13.8 μM (dihydropinosylvin). In HT-29, the values ranged from 24.4 ± 11.3 μM (4-hydroxy-trans-stilbene) to 96.7 ± 6.7 μM (4-hydroxydibenzyl). At their IC50, most compounds induced apoptosis and arrested the cell cycle at the S phase, pterostilbene at G2/M, while 4-hydroxy-trans-stilbene and 3,4\'-dihydroxy-trans-stilbene arrested at both phases. Higher Connolly values (larger size) hindered the antiproliferative activity, while a lower pKa1 enhanced the activity in Caco-2, and higher LogP values (more hydrophobicity) increased the activity in HT-29. Reducing the styrene double bond in stilbenes was the most critical feature in decreasing the antiproliferative activity. These results (i) suggest that gut microbiota metabolism determines the antiproliferative effects of dietary stilbenes. Therefore, RSV consumption might exert different effects in individuals depending on their gut microbiota metabotypes associated with RSV metabolism, and (ii) could help design customized drugs with a stilbenoid and (or) dibenzyl core against colorectal cancer.

Lactic acid bacteria (LAB) naturally inhabiting the digestive tract of honeybees are known for their ability to detoxify xenobiotics. The effect of chlorpyrifos, coumaphos, and imidacloprid on the growth of LAB strains was tested. All strains showed high resistance to these insecticides. Subsequently, the insecticide binding ability of LAB was investigated. Coumaphos and chlorpyrifos were bound to the greatest extent (up to approx. 64%), and imidacloprid to a much weaker extent (up to approx. 36%). The insecticides were detected in extra- and intracellular extracts of the bacterial cell wall. The ability of selected LAB to reduce the cyto- and genotoxicity of insecticides was tested on two normal (ovarian insect Sf-9 and rat intestinal IEC-6) cell lines and one cancer (human intestinal Caco-2) cell line. All strains exhibited various levels of reduction in the cyto- and genotoxicity of tested insecticides. It seems that coumaphos was detoxified most potently. The detoxification abilities depended on the insecticide, LAB strain, and cell line. The detoxification of insecticides in the organisms of honeybees may reduce the likelihood of the penetration of these toxins into honeybee products consumed by humans and may contribute to the improvement of the condition in apiaries and honeybee health.

BACKGROUND: Lactic acid bacteria (LAB), many of which are probiotics, can produce health-promoting metabolites (postbiotics).
OBJECTIVE: To assess the mechanism of antiproliferative action of postbiotics, post-fermentation media (PFM) and cell extracts (CEs) of several strains of LAB were studied against colon (Caco-2), and cervix (HeLa) cancer cell lines, as well as normal intestine (IEC-6) cells, were used as a comparison.
METHODS: Postbiotics of various LAB (n = 39) were screened for their antiproliferative activity. The effect of PFM and CEs on reactive oxygen species (ROS), mitochondrial membrane potential (MMP), ATP production, phosphatidylserine (PS) externalisation, and apoptosis-related caspases 3/7 and 9 activation was assayed.
RESULTS: PFM and CEs showed strong dose-dependent antiproliferative activity against Caco-2 cells, up to 77.8 ± 0.8% and 58.4 ± 1.6% for PFM and CEs, respectively. Stronger inhibitory activity against cancerous (Caco-2 and HeLa) cells than against normal (IEC-6) cells was observed. PFM were more inhibitory than CEs, and both generated oxidative stress in Caco-2 cells. PFM of L. plantarum 0991 and L. brevis 0983 induced apoptosis in Caco-2 cells by the mitochondrial signalling pathway.
CONCLUSIONS: Anticancer activity of PFM and CEs of LAB, as well as the ability of apoptosis induction, is strain-specific.

High levels of persistent contaminants such as microplastics (MPs) and trace organic compounds (TrOCs) in the aquatic environment have become a major threat on the ecosystem and human health. While MP\'s role as a vector of environmental TrOCs is widely discussed in the literature, the corresponding implications of the interaction between these two compounds on human health (i.e., their joint toxic effect) have not been illustrated. Using a TrOCs model (Triclosan, TCS) and primary MPs (polystyrene microbeads), this work evaluates the sorption and desorption potential of TCS and MPs in simulated environmental and cellular conditions, respectively, and estimates the single and joint toxicity of these interactions toward human cells (Caco-2). Surface functionality of the microbeads highly increased their adsorption capacity of TCS, from 2.3 mg TCS for non-functionalized microbeads to 4.6 mg and 6.1 mg TCS per gram of microbeads for amino- and carboxyl-functionalized MPs, respectively. Using non-functionalized MPs, non-specific \"hydrophobic-like\" interactions and π-π interactions dominated the sorption mechanism of TCS; however, the addition of hydrogen interactions between functionalized microbeads and TCS increased the microbeads\' overall sorption capacity. TCS was desorbed from both functionalized and non-functionalized MPs when changing from environmental conditions to cellular conditions. Desorption was found to be dependent on the matrix complexity and protein content as well as microbead functionality. Finally, toxicity tests suggested that while low concentrations of TCS and MPs (separately) have minor toxic effect toward Caco-2 cells, TCS-sorbed MPs at similar concentrations have an order of magnitude higher toxicity than pristine MPs, potentially associated with the close interaction of both MP and TCS with the cells. Overall, this study not only elucidates the role of MPs as a TrOC vector, but also demonstrates a realistic scenario in which co-presence of these environmental contaminants poses risks to the environment and human health.

Ankaferd Blood Stopper (ABS) is a medicinal plant extract that has anti-inflammatory effect. Inflammatory bowel disease is a pathological condition that directly affects colon health and increases the risk of colon cancer. Especially inflammation is an important factor in the formation and progression of this disease. The aim of the study was to investigate the protective effect of ABS on colonic inflammation. Caco-2 and RAW 264.7 cells were used as a model of in vitro colonic inflammation. RAW 264.7 cells were treated with lipopolysaccharide for 12 h to induce inflammation, and an inflammatory medium (IM) was obtained. Caco-2 cells were treated with 15 μL/mL ABS for 4 h, then incubated with IM. The cells also were incubated with 15 μL/mL ABS and IM together for 12 h. Tumor necrosis factor alpha (TNF-α) protein levels were targeted in testing inflammatory condition and cyclooxygenase-2 (COX-2) mRNA level was used as a marker gene to show the possible anti-inflammatory effect of ABS in Caco-2 cells. TNF-α level was 26.1-fold higher than the control group. IM caused 3.2-fold increase in COX-2 expression in Caco-2 cells. Pretreatment of Caco-2 cells with ABS resulted in 3.3-fold decrease in COX-2 mRNA levels relative to IM group. Furthermore, COX-2 mRNA level reduced 4.7-fold when ABS and conditional medium were given at the same time. ABS has suppressive effect on COX-2 mRNA expression in Caco-2 cells. These results suggest that ABS might have protective and therapeutic effect for colonic inflammation.

The present study investigated the effects of foliar application of zinc (Zn) and selenium (Se) on bioavailability of Zn and Se and toxicity of cadmium (Cd) and lead (Pb) to different water spinach ecotypes (LA and HA) grown in slightly (XZ) or moderately (LJY) contaminated fields via in vitro digestion combined with Caco-2/HL-7702 cell model. The obtained results revealed that foliar application of Zn and Se promoted yield, increased total, bioaccessible and bioavailable fractions of Zn and Se in plants, indicating that foliar application is a feasible way of biofortification. Although there was no significant effect on liver cell proliferation (MTT), membrane stability (LDH) and hepatocyte enzyme (ALT and AST) activities, the obvious ecotype and soil dependent fluctuations of lipid peroxidation (MDA) and antioxidant enzyme (SOD, POD and CAT) activities in serum highly suggest that the low accumulator and clean field should be used in agricultural production rather than the high accumulator and contaminated farmland. Moreover, foliar application of Zn and Se improved nutritional quality of all water spinach genotypes in both fields, including increased Fe, vitamin C, cellulose and chlorophyll, maintained concentrations of potassium (K), manganese (Mn), copper (Cu), protein, and nitrate. These results demonstrate that this agricultural management practice may prove to be an effective approach for minimizing health risk and alleviating \"hidden hunger\" in the developing countries.

Flavonoids, including anthocyanins, are polyphenolic compounds present in fruits, vegetables and dietary supplements. They can be absorbed from the intestine to the bloodstream or pass into the large intestine. Various bacterial species and enzymes are present along the entire intestine. The aim of the present work was to investigate the intestinal metabolism of selected dietary polyphenol and polyphenol glycosides (quercetin, cyanidin-3-O-glucoside, cyanidin-3-O-galactoside, and delphinidin-3-O-galactoside) by human fecal bacteria. Moreover, the metabolism of metabolites formed from these compounds in human colon carcinoma cells (Caco-2) was also point of the interest. Test compounds were added to fresh human stool in broth or to Caco-2 cells in medium and then incubated for 6 or 20 h at 37°C. After incubation, samples were prepared for LC/MS determination. Main metabolic pathways were deglycosylation, hydrogenation, methylation, hydroxylation, and decomposition. 2,4,5-trihydroxybenzaldehyde, as a metabolite of cyanidin glycosides, was detected after incubation for the first time. Metabolites formed by fecal bacteria were further glucuronidated or methylated by intestinal enzymes. This metabolite profiling of natural compounds has helped to better understand the complex metabolism in the human intestine and this work also has shown the connection of metabolism of natural substances by intestinal bacteria followed by metabolism in intestinal cells.