Human Colon Cancer Caco-2 Research Articles

Antioxidant and pro-apoptosis activities of coffee husk (Coffea arabica) anthocyanins

The commercial use of coffee (Coffea arabica) husks, one of the major solid residues obtained during the dry processing of coffee, has previously been considered unprofitable. However, coffee husks are an excellent source of stable anthocyanins that have antioxidative, anti-inflammatory, and cardioprotective properties. The present work identified two anthocyanins, cyanidin-3-glucoside and cyanidin-3-rutinoside, from coffee husks by high-performance liquid chromatography and liquid chromatography-mass spectrometry. The antioxidant capabilities were tested by quenching free radical scavenge assay, reducing power, and ORAC assay at 50, 100, 150, 200 mg/L of coffee husk anthocyanins (CHAs). Moreover, the effects of different concentrations of CHAs on superoxide dismutase (SOD) and lactate acid dehydrogenase (LDH), and the concentration of malondialdehyde (MDA) in human umbilical vein endothelial cells exposed to hydrogen peroxide were also evaluated. Human colon cancer (Caco-2) cell apoptosis induced by CHAs was examined by flow cytometry. Based on the results, CHAs showed strong dose-dependent antioxidant activities, and could increase SOD activity, and suppress indicators for oxidative injuries such as MDA and LDH. Furthermore, the proportion of apoptotic Caco-2 cells ranged from 4.12 to 41.3% in response to treatment with different concentrations of CHAs. These results suggest that CHAs exhibit antioxidant and pro-apoptosis activities.

Enhancement of Apo2L/TRAIL signaling pathway receptors by theactivation of Klotho genewithCRISPR/Cas9 in Caco-2 colon cancer cells.

Human Klotho gene has many known functions such as anti-aging and anti-tumor, and decreased expression of this gene causes malignant formations in most types of cancer, including colon cancer. Interacting with TRAIL death receptors (DR4 and DR5) induces an apoptotic effect in cancer treatments by reducing the proliferation of cancer cells. The present study aimed to investigate downstream effect of overexpression of Klotho gene, which is known to have an antitumor effect on resistant human colon cancer cells, by examining its action on TRAIL death and decoy (DcR1 and DcR2) receptors for the first time. For this purpose, upregulation of human Klotho gene was achieved with CRISPR/Cas9-mediated system in resistant human colon cancer Caco-2 cells. To determine the effect of upregulation of Klotho gene on cancer cells evaluations with flow cytometry, WST-8, qRT-PCR, ELISA, and immunohistochemical analysis were performed. Then, Klotho gene was knocked out and its apoptotic effect was tested to find out whether it is due to overexpression of Klotho gene or not. Our results indicate that overexpression of Klotho gene in Caco-2 cells via CRISPR/Cas9-sensitized TRAIL death receptor DR4 suppresses the proliferation of cells by leading to apoptosis. Thus, this study conducted on apoptosis-resistant colon cancer cells may bring new insights about the role of Klotho gene in colon cancer.

Assessment of the in vitro anticancer activities of cyanobacteria mediated silver oxide and gold nanoparticles in human colon CaCo-2 and cervical HeLa cells

This work aims to assess the cytotoxicity of silver oxide and gold nanoparticles on two cancer cell cultures, human colon CaCo-2, and cervical HeLa cancer cells. Silver oxide and gold nanoparticles were synthesized using the isolated microalgae Oscillatoria sp. and Spirulina platensis. Different characterization tools (UV–Vis spectrum, XRD diffraction, TEM photography, FTIR spectroscopy, and Zeta-potential) had been used to confirm the resulted nanoparticles. Thereafter, the resulted silver oxide and gold nanoparticles were assessed as anticancer agents against human colon cancer (CaCo-2), cervical cancer (HeLa), and normal cells (WISH). The effective interaction of Oscillatoria sp. mediated Ag2O|AgO-NPs and S. platensis mediated Au-NPs against the normal human cells (WISH) and the cancer cells (CaCo-2 and HeLa), as IC50 of Ag2O|AgO-NPs recorded 147.77, 252.83, and 286.74 μg/ml for WISH, CaCo-2, and HeLa cells, respectively. IC50 for Au-NPs recorded 307.31, 311.00, and 382.90 μg/ml for WISH, CaCo-2, and HeLa cells, respectively. The safety limit against HeLa reached 98.61% by 3.906 μg/ml of Ag2O|AgO-NPs, and it recorded 93.13% by 15.625 μg/ml against CaCo-2, while the results of WISH was 51.29 % by 125.00 μg/ml. Concerning the anticancer effect of Au-NPs, the safety limit recorded 92.47% by 3.906 μg/ml against CaCo-2, while HeLa reached 93.43% by 15.625 μg/ml and WISH was 99.62% by 62.500 μg/ml. The cytotoxicity results revealed that both Oscillatoria sp. mediated Ag2O|AgO-NPs and S. platensis mediated Au-NPs caused the reduction of the colon (CaCo-2) and cervical cancer cell lines. Although the low concentrations of silver oxide nanoparticles 3.906 and 15.625 μg/ml showed the safety limit effect against HeLa and CaCo-2, they revealed no hazard on WISH cells. In conclusion, algal silver oxides and gold nanoparticles exhibited effective anticancer activities against cancer cells as a green nanotechnology application.

Ketogenesis alleviates TNFα-induced apoptosis and inflammatory responses in intestinal cells

The disturbance of strictly regulated self-regeneration in mammalian intestinal epithelium is associated with various intestinal disorders, particularly inflammatory bowel diseases (IBDs). TNFα, which plays a critical role in the pathogenesis of IBDs, has been reported to inhibit production of ketone bodies such as β-hydroxybutyrate (βHB). However, the role of ketogenesis in the TNFα-mediated pathological process is not entirely known. Here, we showed the regulation and role of HMGCS2, the rate-limiting enzyme of ketogenesis, in TNFα-induced apoptotic and inflammatory responses in intestinal epithelial cells. Treatment with TNFα dose-dependently decreased protein and mRNA expression of HMGCS2 and its product, βHB production in human colon cancer cell lines HT29 and Caco2 cells and mouse small intestinal organoids. Moreover, the repressed level of HMGCS2 protein was found in intestinal epithelium of IBD patients with Crohn's disease and ulcerative colitis as compared with normal tissues. Furthermore, knockdown of HMGCS2 enhanced and in contrast, HMGCS2 overexpression attenuated, the TNFα-induced apoptosis and expression of pro-inflammatory chemokines (CXCL1-3) in HT29, Caco2 cells and DLD1 cells, respectively. Treatment with βHB or rosiglitazone, an agonist of PPARγ, which increases ketogenesis, attenuated TNFα-induced apoptosis in the intestinal epithelial cells. Finally, HMGCS2 knockdown enhanced TNFα-induced reactive oxygen species (ROS) generation. In addition, hydrogen peroxide, the major ROS contributing to intestine injury, decreased HMGCS2 expression and βHB production in the intestinal cells and mouse organoids. Our findings demonstrate that increased ketogenesis attenuates TNFα-induced apoptosis and inflammation in intestinal cells, suggesting a protective role for ketogenesis in TNFα-induced intestinal pathologies.

Cytotoxicity of Mongolian Medicine <i>Aconitum kusnezoffii</i>

Aconitum kusnezoffii used as an analgesic and anti-inflammatory. It is often used not only in Mongolian medicine but also in traditional Chinese medicine. However, there are few studies on its impact of inflammatory cytokines. This study explored the effects of Aconitum kusnezoffii in vitro used human colon cancer (Caco-2) cells. Cytotoxicity was assessed by measuring effects on expression of IL-1β, IL-6, IL-8, and TNF-α used enzyme-linked immunosorbent assay (ELISA). In vitro study demonstrated that incubation of Caco-2 cells with some kinds of Aconitum kusnezoffii at some concentrations inhibit secretion of TNF-α and FAXIU ACONITUM KUSNEZOFFII could inhibit the secretion of IL-1β. While IL-6 and IL-8 levels were unaffected after 24 h of treatment with Aconitum kusnezoffii, qujian Aconitum kusnezoffii, faxiu Aconitum kusnezoffii.

A specific selenium-chelating peptide isolated from the protein hydrolysate of Grifola frondosa.

Background: Grifola frondosa is a type of edible medicinal mushroom with abundant proteins. Selenium (Se) is an essential micronutrient for human. Many animal experiments and clinical studies had indicated that Se plays an important role in diverse physiologic actions. Most inorganic selenium compounds are toxic, and the lowest lethal dose is relatively small. Peptide-Se chelate can probably be dietary supplements in functional foods for humans with Se deficiency. Methods: In this study, a specific tripeptide Arg-Leu-Ala (RLA) with strong Se-chelating capacity was purified from Grifola frondosa through ultrafiltration, reversed-phase HPLC and gel filtration chromatography. The UV, SEM, XRD, 1H NMR spectra are shown to provide more information about characterization of RLA-Se chelates. The bioavailability of RLA-Se chelate in Caco-2 cell line was investigated by using human colon cancer Caco-2 cells as model. iTRAQ comparative proteomics approach were used to identify the differentially expressed proteins. Results: The Se binding capacity of RLA was 84.47 ± 1.21 mg g−1. The results of UV, X-ray diffraction (XRD), 1H NMR and SEM structure analysis showed that the binding of selenium in the hydrolysate of Grifola frondosa protein was successful, and the amino and carboxyl groups of RLA were involved in the coordination of Se, which was the main site of chelation. The results of absorption of RLA-Se chelate in Caco-2 cells showed that RLA-Se chelate could be used as selenium supplement source. Using iTRAQ comparative proteomics approach, 40 proteins found significant. RLA-Se treatment had been demonstrated to present a higher accumulation of Se compared with control treatment and show an effective absorption by Caco-2 with the result that E3 protein performed up regulation. RLA-Se may play roles in cell cycle and apoptosis as an essential micronutrient. To sum up, our research results show that Grifola polypeptide-Se chelate is a promising multifunctional organic selenium product, which can be used as a new functional supplement for selenium deficiency.

Bivalence metal complexes of antithyroid drug carbimazole; synthesis, characterization, computational simulation, and biological studies

New metal complexes of carbimazole (CMZ) were synthesized with a molar ratio of 1:2. The bivalence complexes were characterized by various techniques, such as elemental analyses and UV–visible spectra, magnetic, and thermal analysis. The metal-ligand stability constant obtained from the molar ratio experiment showed high copper complex stability among the prepared metal compounds at room temperature (Kf =6.6 × 105). The infrared spectra (IR) showed that the CMZ ligand coordinated to the metal through the sulfur atom and the oxygen ether group. The spectroscopic results suggested tetrahedral geometry for all complexes except Ni(II) and Zn(II) complexes with a square planar arrangement. The proposed structures were optimized using density functional theory (DFT/B3LYP) and 6–311G*(d,p) basis sets. The geometrical optimization parameters and molecular electrostatic potentials were calculated. Biological investigation started with DNA binding spectroscopy, which revealed a high binding ability of the Cu(II) and Ni(II) complexes to the CT-DNA. A docking study using different protein receptors was implemented to study the proposed drug's molecular behavior. The in vitro cytotoxicity evaluation of the complexes toward breast human cancer (Mcf7) and human colon cancer (CaCo-2) cell linesindicated that the Cu(II)-CMZ complex has high potential anticancer activities against those cell lines.

Characterization and biological activities of synthesized zinc oxide nanoparticles using the extract of Acantholimon serotinum

AbstractThe present study reports the synthesis of ZnO-NPs using Acantholimon serotinum extracts followed by characterization and evaluation of biological activities. Field emission scanning electron microscope revealed irregular spherical morphology with a size in the range of 20–80 nm. The X-ray diffraction analysis confirmed the synthesis of highly pure ZnO NPs with a hexagonal shape and a crystalline size of 16.3 nm. The UV-Vis spectroscopy indicates the synthesis of ZnO-NPs. FT-IR confirmed the presence of phytocomponents in the plant extract, which was responsible for nanoparticle synthesis. According to MTT results, the biosynthesized ZnO-NPs showed cytotoxic effects on human colon cancer Caco-2 (IC50: 61 µg/mL), neuroblastoma SH-SY5Y (IC50: 42 µg/mL), breast cancer MDA-MB-231 (IC50: 24 µg/mL), and embryonic kidney HEK-293 (IC50: 60 µg/mL) cell lines. Significant reactive oxygen species (ROS) generation was measured by the DCFH-DA assay after 24 h incubation with ZnO-NPs (200 µg/mL). ZnO-NPs caused apoptotic and necrotic effects on cells, which was confirmed by Annexin V-PE/7-AAD staining and 6.8-fold increase in pro-apoptosis gene Bax and 178-fold decrease in anti-apoptosis gene Bcl-2. The well diffusion method did not show effective growth inhibition activities of the ZnO-NPs against bacteria. In conclusion, the ZnO-NPs induce cytotoxicity in cell lines through ROS generation and oxidative stress.

Polysaccharide fractions from Handroanthus heptaphyllus and Handroanthus albus barks: Structural characterization and cytotoxic activity

Barks of trees of the genus Handroanthus are known for their antitumor activity, which is attributed to naphthoquinones. Another class of molecules that has shown antitumor activity are the polysaccharides, however those from Handroanthus barks have never been studied. Accordingly, the aim of this study was to extract polysaccharides from H. heptaphyllus and H. albus barks, to characterize them structurally and to evaluate their cytotoxic effects on the human colon and human breast cancer cell lines, Caco-2 and MCF-7, respectively. The polysaccharides were extracted with boiling water and fractionated by freeze-thawing process. The soluble polysaccharide fractions HHBSF and HABSF were characterized by monosaccharide composition, methylation and NMR analyses, and their effects on proliferation of Caco-2 and MCF-7 cells were evaluated using MTT cell viability assay. HHBSF and HABSF were mainly constituted of galactoglucomannan, type II arabinogalactan (AGII) and type I rhamnogalacturonan (RGI), however, only HABSF significantly inhibited the growth of MCF-7 (CC50 = 327 μg/mL) and Caco-2 (CC50 = 2258 μg/mL) cells. Differences in the fine structure and proportion of their polysaccharides, and maybe in the composition of associated phenolic compounds could explain the different effects of HHBSF and HABSF.

Preparation and characterisation of bifunctional surface-modified silicone catheter in lumen

ObjectivesThe purpose of this study was to evaluate the coating of antimicrobial peptides (AMPs) and polyvinylpyrrolidone (PVP) to the surface of a silicone catheter to reduce bacterial growth and to increase hydrophilicity, respectively. MethodsSurface characterisation was performed on bare silicone, AMP-coated, PVP-coated and AMP + PVP-coated silicone catheters using attenuated total reflectance-infrared (ATR-IR) spectroscopy, X-ray photoelectron spectroscopy (XPS) and water contact angle. Antibacterial activity, antibacterial biofilm growth and XTT assay were performed on bare silicone, AMP-coated, PVP-coated and AMP + PVP-coated silicone catheters. Statistical analysis was performed by one-way ANOVA. ResultsThe water contact angle of the AMP + PVP-coated silicone catheter was 21.37 ± 2.17° compared with 107.23 ± 0.96°, 74.40 ± 1.76° and 20.77 ± 0.32° for bare silicone, AMP-coated and PVP-coated silicone catheters. Based on in vitro antimicrobial tests, the AMP + PVP-coated silicone catheter had 6.2, 2.2 and 2.5 greater antibacterial activity than that of the bare silicone catheter against Escherichia coli, Staphylococcus aureus and Pseudomonas aeruginosa, respectively. Moreover, bacterial biofilm growth on the surface of the AMP + PVP-coated silicone catheter was minimal as characterised by scanning electron microscopy. MTT assay showed that bare silicone, AMP-coated, AMP + PVP-coated and PVP-coated silicone catheters were non-cytotoxic to 3T3 and human colon cancer (Caco-2) cells. ConclusionsThis work demonstrates that AMP + PVP-coated silicone catheters have potential clinical application prospects with improved hydrophilicity, excellent biocompatibility, antibacterial activity and a certain antibacterial biofilm effect.

In-vitro anticancer activity against Caco-2 cell line of colloidal nano silver synthesized using aqueous extract of Eucalyptus Camaldulensis leaves

In the current study, we investigated the anticancer potential against human colon cells (Caco-2) of colloidal nanosilver (CN–Ag) produced in Syria using bioactive compounds in the aqueous extract of Eucalyptus camaldulensis leaves (AEECL). The formation of AgNPs was confirmed by UV-visible spectroscopy analysis with surface plasmon peak at 449 nm and their average size was found to be 12, 10, 23 nm by SEM, DLS and NTA respectively. This small size has confirmed the effective role of AEECL as capping agent. Further morphological characterization was done by EDS showed the presence of metallic silver. Zeta potential value (-23 mV) indicated the repulsion among the particles and stability of the formulation nanosilver. The anticancer effect of synthesized CN–Ag against Caco-2 has been tested. The cytotoxicity assay showed a dose-dependent and a time-dependent effect of CN–Ag. The high cytotoxicity of CN–Ag at low concentration (5μ/mL) open new prospects for the development of novel therapeutic approaches against human colon cancer Caco-2.

Design synthesis and anti-proliferative activity of some new coumarin substituted hydrazide–hydrazone derivatives

A series of 21 coumarin hydrazide–hydrazone derivatives were designed, synthesized and evaluated potential cytotoxicity effects at 25 μg/mL for 48 h against liver cancer (HepG2) cell line in vitro. Then, seven out of 21 compounds with % cell viability lower than 60% were selected for evaluation of in vitro anti-proliferative activity against liver cancer (HepG2), breast cancer (SKBR-3) and human colon cancer (Caco-2) cell lines. Among the test compounds, 5g, 6d and 6f showed potent activities against both Hep-G2 and SKBR-3 cell lines. More significantly, compound 6d, having a 4-bromophenyl moiety, exhibited best cytotoxic activity against Hep-G2 cell line with IC50 value of 2.84 ± 0.48 μg/mL which is comparable to the standard doxorubicin (IC50 = 2.11 ± 0.13 μg/mL). In addition, compound 6f, having 4-methoxyphenyl moiety, demonstrated the most potent activity (IC50 = 2.34 ± 0.68 μg/mL) against SKBR-3 cell line on comparison with other tested coumarin hydrazide–hydrazone derivatives. Unfortunately, all test compounds, as well as doxorubicin, showed no cytotoxicity toward drug-resistant cell line, Caco-2. Our preliminary results indicated that coumarin hydrazide–hydrazone derivatives could be exploited as leading structures for further anticancer-drug development.

Antifungal and cytotoxic activities of acetone and aqueous extracts of Morinda lucida

Aspergillosis is an opportunistic airborne fungal infection caused by ubiquitous saprophytic moulds of the Aspergillus genus. Infection has been documented in humans and animals, and it usually occurs through inhalation of unicellular spores known as conidia or, more rarely, after ingestion or wound contamination. Oral candidiasis is also an important sign for clinical diagnosis and an indicator of the evolution of immunodeficiency among HIV carriers. Mycotic stomatitis in animals is caused by Candida spp. in most cases. Cryptococcosis is a fungal disease of man and domestic animals, caused by the Cryptococcus neoformans complex. In animals it is often characterized by upper respiratory symptoms, subcutaneous nodules, pneumonia, central nervous system or ocular disorders, lymphadenopathy, or subcutaneous nodules. Antifungal drugs are often unaffordable to rural farmers and their efficacy is hampered as a result of antimicrobial resistance. Therefore, the need to seek alternative measures from phytogenic sources is urgent. The aim of this study was to investigate the in vitro antifungal activity of acetone and aqueous extracts of Morinda lucida (Rubiaceae) against ATCC strains of Aspergillus fumigatus, A. flavus, Candida albicans and Cryptococcus neoformans as well as clinical isolates of A. fumigatus and C. albicans using a serial microdilution assay. The cytotoxicity against human colon cancer (Caco‐2) cells and total activity of the acetone and aqueous extracts were also determined. The minimum inhibitory concentration (MIC) of both extracts against tested organisms ranged from 0.11 to 2.50 mg/ml and 0.03 to 2.50 mg/ml after 48 and 72 hours respectively. The LC50 values of the acetone and aqueous extracts against the Caco‐2 cells were 0.46 and 0.58 mg/ml respectively. The range of selectivity index (SI) values of both extracts was 0.23 to 5.27 and 0.23 to 19.33 after 48 and 72 hours respectively. The range of total activity of the extracts was 56 to 2 454 and 56 to 9 000 ml/g after 48 and 72 hours respectively. The potential of this plant species as an alternative for treatment of human and livestock mycoses is supported by these results. However, in vivo data is necessary to further investigate this claim

Cytotoxicity, Cell Cycle Arrest, and Apoptosis Induction Activity of Ethyl-p-methoxycinnamate in Cholangiocarcinoma Cell.

Objective:To investigate cytotoxic activity of ethyl-p-methoxycinnamate (EPMC) including its effect on p-glycoprotein (multidrug resistance-1: mdr-1 gene) in human cholangiocarcinoma cell. Methods:Cytotoxic activity of EPMC against human cholangiocarcinoma (CL-6), fibroblast (OUMS-36T-1F), and colon cancer (Caco-2) cell lines were assessed using MTT assay. Selectivity index (SI) was determined as the ratio of IC50 (concentration that inhibits cell growth by 50%) of EPMC in OUMS-36T-1F and that in CL-6 cell. Cell cycle arrest and apoptosis in CL-6 cells were investigated by flow cytometry and fluorescent microscopy. Effect of EPMC on mdr-1 gene expression in CL-6 and Caco-2 was determined by real-time PCR. Results:The median (95% CI) IC50 values of EPMC in CL-6, OUMS-36T-1F, and Caco-2 were 245.5 (243.1-266.7), 899.60 (855.8-966.3) and 347.0 (340.3-356.9) µg/ml, respectively. The SI value of the compound for the CL-6 cell was 3.70. EPMC at IC50 inhibited CL-6 cell division and induced apoptosis compared to untreated control. EPMC exposure did not induce mdr-1 gene expression in both CL-6 and Caco-2 cells. Conclusion:The results suggest the potential role of EPMC in cholangiocarcinoma with a low possibility of drug resistance induction.

RASAL1 induces to downregulate the SCD1, leading to suppression of cell proliferation in colon cancer via LXR\u03b1/SREBP1c pathway

BackgroundRecent studies have confirmed that RASAL1 has an antitumor effect in many cancers, but its functional role and the molecular mechanism underlying in colon cancer has not been investigated.ResultsWe collected human colon cancer tissues and adjacent non-tumor tissues, human colon cancer cell lines LoVo, CaCo2, SW1116, SW480 and HCT-116, and normal colonic mucosa cell line NCM460. RT-qPCR was used to detect the RASAL1 level in the clinical tissues and cell lines. In LoVo and HCT-116, RASAL1 was artificially overexpressed. Cell viability and proliferation were measured using CCK-8 assays, and cell cycle was detected via PI staining and flow cytometry analysis. RASAL1 significantly inhibited the cell proliferation via inducing cell cycle arrest, suppressed cell cycle associated protein expression, and decreased the lipid content and inhibited the SCD1 expression. Moreover, SCD1 overexpression induced and downregulation repressed cell proliferation by causing cell cycle arrest. Additionally, luciferase reporter assays were performed to confirm the direct binding between SREBP1c, LXRα and SCD1 promoter, we also demonstrated that RASAL1 inhibit SCD1 3′-UTR activity. RASAL1 inhibited tumor growth in xenograft nude mice models and shows inhibitory effect of SCD1 expression in vivo.ConclusionTaken together, we concluded that RASAL1 inhibited colon cancer cell proliferation via modulating SCD1 activity through LXRα/SREBP1c pathway.

MiR-34b inhibits the proliferation and promotes apoptosis in colon cancer cells by targeting Wnt/β-catenin signaling pathway.

Colon cancer is one of the leading cause of cancer deaths that is severely threatening human health. Several microRNAs (miRNAs) have been found to be associated with the tumor genesis of colon cancer. The present study determined the expression of miR-34b in patients with colon cancer and studied the molecular mechanism of miR-34b in the proliferation and apoptosis of human colon cancer Caco-2 cells in vitro. In colon cancer patients, the expression of miR-34b was decreased in tumor tissues when compared with the adjacent non-tumor tissues. Furthermore, overexpression of miR-34b inhibited proliferation, migration and invasion, while promoted apoptosis in colon cancer cells. The online bioinformatics sites predicted possible regulatory genes of miR-34b and luciferase reporter assay verify that β-catenin was a direct target of miR-34b. Furthermore, miR-34b overexpression significantly decreased the expression of genes associated with Wnt/β-catenin signaling pathway. In conclusion, our results suggest that miR-34b may inhibit migration and invasion of human colon cancer cells by regulating Wnt/β-catenin signaling and miR-34b may be a key target for the treatment and diagnosis of colon cancer.

Regulation of Ketogenic Enzyme HMGCS2 by Wnt/β-catenin/PPARγ Pathway in Intestinal Cells.

The Wnt/β-catenin pathway plays a crucial role in development and renewal of the intestinal epithelium. Mitochondrial 3-hydroxy-3-methylglutaryl-CoA synthase 2 (HMGCS2), a rate-limiting ketogenic enzyme in the synthesis of ketone body β-hydroxybutyrate (βHB), contributes to the regulation of intestinal cell differentiation. Here, we have shown that HMGCS2 is a novel target of Wnt/β-catenin/PPARγ signaling in intestinal epithelial cancer cell lines and normal intestinal organoids. Inhibition of the Wnt/β-catenin pathway resulted in increased protein and mRNA expression of HMGCS2 and βHB production in human colon cancer cell lines LS174T and Caco2. In addition, Wnt inhibition increased expression of PPARγ and its target genes, FABP2 and PLIN2, in these cells. Conversely, activation of Wnt/β-catenin signaling decreased protein and mRNA levels of HMGCS2, βHB production, and expression of PPARγ and its target genes in LS174T and Caco2 cells and mouse intestinal organoids. Moreover, inhibition of PPARγ reduced HMGCS2 expression and βHB production, while activation of PPARγ increased HMGCS2 expression and βHB synthesis. Furthermore, PPARγ bound the promoter of HMGCS2 and this binding was enhanced by β-catenin knockdown. Finally, we showed that HMGCS2 inhibited, while Wnt/β-catenin stimulated, glycolysis, which contributed to regulation of intestinal cell differentiation. Our results identified HMGCS2 as a downstream target of Wnt/β-catenin/PPARγ signaling in intestinal epithelial cells. Moreover, our findings suggest that Wnt/β-catenin/PPARγ signaling regulates intestinal cell differentiation, at least in part, through regulation of ketogenesis.

Electrospun oral formulations for combined photo-chemotherapy of colon cancer

In this work, we report new formulations for the combined photo-chemotherapy of colon cancer. Fibers were fabricated via coaxial-electrospinning with the intent of targeting delivery of the anti-cancer drug carmofur (CAR) and the photosensitizer rose bengal (RB) selectively to the colon site. The fibers comprised a hydroxypropyl methylcellulose (HPMC) core loaded with the active ingredients, and a pH-sensitive Eudragit L100-55 shell. The fibers were found to be homogeneous and cylindrical and have visible core-shell structures. X-ray diffraction and differential scanning calorimetry demonstrated that both CAR and RB were present in the fibers in the amorphous physical form. In vitro drug release studies showed that the fibers have the potential to selectively deliver drugs to the colon, with only 10–15 % release noted in the acidic conditions of the stomach but sustained release at pH 7.4. Cytotoxicity studies were undertaken on human dermal fibroblast (HDF) and colon cancer (Caco-2) cells, and the influence of light on cell death was also explored. The fibers loaded with CAR alone showed obvious toxicity to both cell lines, with and without the application of light. The RB-loaded fibers led to high viability (ca. 80% for both cell types) in the absence of light, but much greater toxicity was noted (30–50%) with light. The same trends were observed with the formulation containing both CAR and RB, but with lower viabilities. The RB and RB/CAR loaded systems show clear selectivity for cancerous over non-cancerous cells. Finally, mucoadhesion studies revealed there were strong adhesive forces between the rat colonic mucosa and the fibers after they had passed through an acidic environment. Such electrospun fibers thus could have potential in the development of oral therapies for colon cancer.

Tumor suppressive effects of the pleiotropically acting miR-195 in colorectal cancer cells.

Downregulation of miR-195 in colorectal cancer tissues has been reported in several studies. We investigated the impact exogenous induction of mature miR-195-5p on some malignant features of human colorectal cancer cells. Caco-2 and SW480 human colon cancer cell lines were transfected with a synthetic miR-195-5p mimic. Exogenous induction of miR-195-5p suppressed multiple mediators of invasion and angiogenesis in colorectal cancer cells and increased the apoptotic cell population in both cell lines. Also, migration of both cell lines was significantly compromised after miR-195 transfection. Our results are indicating a strong tumor suppressive role for miR-195 in human colorectal cancer.

Biosynthesis of copper nanoparticles using aqueous Tilia extract: antimicrobial and anticancer activities

A cost-effective method for the biosynthesis of copper nanoparticles (Cu-NPLs) using Tilia extract under optimum conditions has been presented. The use of Tilia extracts for the synthesis of Cu-NPLs has been investigated for the first time. The Cu-NPLs are stable due to in situ bio-capping by the Tilia extract residues. Formation of metallic Cu was revealed by UV-vis and XRD analyses. UV-vis of Cu-NPLs showed an SPR characteristic peak at 563 nm (energy bandgap = 2.1 eV). Morphology and size of the as-prepared Cu-NPLs were determined using SEM and TEM studies. TEM observations show that the produced Cu-NPLs are hemispherical in shape with different diameters in the range 4.7–17.4 nm. The electrical conductivity of the Cu-NPLs was determined as 1.04 × 10−6 S cm-1 (at T = 120 K). The antimicrobial studies exhibited relatively high activity against pathogenic bacteria like Gram-positive & Gram-negative bacteria. Anticancer studies demonstrated the in vitro cytotoxicity value of Cu-NPLs against tested human colon cancer Caco-2 cells, human hepatic cancer HepG2 cells and human breast cancer Mcf-7 cells. To conclude, Cu-NPLs are promising in electronic devices and they possess a potential anticancer application for some human cancer therapy as well.