Human Colon Cancer Caco-2 Cells Research Articles

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.

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.

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.

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.

Curcumin and silver nanoparticles carried out from polysaccharide-based hydrogels improved the photodynamic properties of curcumin through metal-enhanced singlet oxygen effect.

Colorectal cancer (CRC) has a high incidence and resistance to conventional treatments. Curcumin (CUR) is a promising natural product in the treatment of CRC with excellent in vitro results. However, its low bioavailability is a limiting factor in clinical applications. To overcome, CUR was incorporated into hydrogels constituted by chitosan (CHT) and chondroitin sulfate (CS), natural biopolymers, capable of controlled release. Hydrogels were synthesized in ionic liquids (ILs, [Hmim][HSO4]) improving the solubility of CHT and the hydrogel properties. Furthermore, CUR was combined with silver nanoparticles (AgNPs) and visible light by Photodynamic Therapy (PDT), which, through the MEO effect (Metal-Enhanced Singlet Oxygen), leads to cell death. It is highlighted the green synthesis of AgNPs using an ultrasound bath. The CHT/CS hydrogels loaded with CUR/AgNPs were properly characterized. Cellular assays showed that the hydrogels (CHT/CS) were not cytotoxic to healthy tissues. However, PDT selective illumination led to inhibition of Caco-2 human colon cancer cells by the CHT/CS/CUR-AgNPs (CC50=91.5μgmL-1 of hydrogel). The cellular uptake assays showed, in addition to the therapeutic action, that the CUR can works as a diagnostic fluorescence probe (theranostic system). Finally, we highlight our commitment to work with reagents, solvents, and methodologies aiming at the principles of green chemistry.

Stearoyl-CoA Desaturase-1 Enzyme Inhibition by Grape Skin Extracts Affects Membrane Fluidity in Human Colon Cancer Cell Lines.

The polyphenolic compounds present in grape extracts have chemopreventive and anticancer properties. Here, we studied the ability of two grape skin extracts (GSEs), Autumn Royal and Egnatia, to influence the cell motility and membrane fluidity regulated by the enzyme Stearoyl-CoA desaturase-1 (SCD1) which increases with the cancer aggressiveness. Caco2 and SW480 human colon cancer cell lines were treated with increasing concentrations of GSEs to evaluate cell proliferation and motility. SCD1 levels were evaluated in both treated cell lines, by membrane lipidomic analysis conducted by gas chromatography. The expression levels of SCD1 and other factors involved in the reorganization of the cytoskeleton and focal adhesions were assessed by Real-time PCR, Western Blotting, and Immunofluorescence staining. High-performance liquid chromatography (HPLC) analyses were performed to determine the phenolic composition in the GSEs, finding them more expressed in Autumn Royal than in Egnatia. Both treatments reduced the levels of SCD1, phospho-Rac1/Cdc42/Rac1/Cdc42 ratio, Cofilin, Vimentin, and phospho-Paxillin especially in Caco2 compared to SW480, showing a different behavior of the two cell lines to these natural compounds. Our findings show that GSEs block the cell migration and membrane fluidity through a new mechanism of action involving structural cellular components.

Lipid-rich extract from Mexican avocado (Persea americana var. drymifolia) induces apoptosis and modulates the inflammatory response in Caco-2 human colon cancer cells

Abstract Colorectal cancer (CRC) is the third most common type of cancer worldwide. Chemotherapy used in CRC patients has severe side effects. As an alternative, the search to identify natural compounds increases the opportunity to select therapeutics. In this work, the cytotoxic effect of a lipid extract (LEAS) from Mexican native avocado seed (Persea americana var. drymifolia) on colon cancer cells Caco-2 was determined. LEAS was cytotoxic towards Caco-2 cells (IC50 = 28 µg/ml), inducing apoptosis through the activation of caspases 8 and 9. Also, LEAS induced loss of membrane mitochondrial potential, inhibited fatty acid oxidation and increased the superoxide production and mitochondrial ROS. Furthermore, LEAS stimulated secretion of cytokines IL-6 (~500%), IL-8 (~400%) and IL-10 (~150%); whereas IL-1β secretion was inhibited (~50%). The results suggest that LEAS induces apoptosis on Caco-2 cells, indicating that avocado is a source of functional food products that can reduce the risk for development of cancer.

Synergistic Effect of Epigenetic Inhibitors Decitabine and Suberoylanilide Hydroxamic Acid on Colorectal Cancer In vitro.

Colorectal Cancer (CRC) is a common cause of oncological deaths worldwide. Alterations of the epigenetic landscape constitute a well-documented hallmark of CRC phenotype. The accumulation of aberrant DNA methylation and histone acetylation plays a major role in altering gene activity and driving tumor onset, progression and metastasis. In this study, we evaluated the effect of Suberoylanilide Hydroxamic Acid (SAHA), a panhistone deacetylase inhibitor, and Decitabine (DAC), a DNA methyltransferase inhibitor, either alone or in combination, on Caco-2 human colon cancer cell line in vitro. Our results showed that SAHA and DAC, separately, significantly decreased cell proliferation, induced apoptosis and cell cycle arrest of Caco-2 cell line. On the other hand, the sequential treatment of Caco-2 cells, first with DAC and then with SAHA, induced a synergistic anti-tumor effect with a significant enhancement of growth inhibition and apoptosis induction in Caco-2 cell line as compared to cells treated with either drug alone. Furthermore, the combination therapy upregulates protein expression levels of pro-apoptotic proteins Bax, p53 and cytochrome c, downregulates the expression of antiapoptotic Bcl-2 protein and increases the cleavage of procaspases 8 and 9; this suggests that the combination activates apoptosis via both the intrinsic and extrinsic pathways. Mechanistically, we demonstrated that the synergistic anti-neoplastic activity of combined SAHA and DAC involves an effect on PI3K/AKT and Wnt/β-catenin signaling. In conclusion, our results provide evidence for the profound anti-tumorigenic effect of sequentially combined SAHA and DAC in the CRC cell line and offer new insights into the corresponding underlined molecular mechanism.

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.

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.

Antigenotoxicity and Cytotoxic Potentials of Metabiotics Extracted from Isolated Probiotic, Lactobacillus rhamnosus MD 14 on Caco-2 and HT-29 Human Colon Cancer Cells

Probiotics, the beneficial bacteria produce active metabolites which could probably mimic their anticancer effect and prevent the risk associated with live bacteria. Thus, the study was designed to isolate effective lactic acid bacteria (LAB) and monitor anticancerous potential of their metabiotic extracts. Probiotics were isolated from different sources and their cell free supernatants (CFS) were screened for antigenotoxic and cytotoxic potentials using SOS Chromo Test and MTT assay on Caco-2 and HT-29 cells. Organic extracts of CFS were prepared and dissolved in different solvents. The isolate with most effective metabiotic extract in terms of cytotoxicity was classified for probiotic and phylogenetic characters and the metabiotic extract was characterized physiochemically. Among 60 isolated LAB, CFS of only 10 isolates showed antigenotoxicity more than 30% and four exhibited 70–80% cytotoxicity. Further, organic extracts of these four CFS dissolved in carboxymethyl cellulose showed 80–90% cytotoxicity. Interestingly, the most effective isolate was found to possess probiotic attributes and phylogenetic characterization revealed it to be Lactobacillus rhamnosus MD 14. Physiochemical characterization of its metabiotic extract indicated the presence of heat sensitive organic acids and proteins. To conclude, metabiotics produced by isolated probiotic L. rhamnosus MD 14 exhibited both antigenotoxic and cytotoxic potential against 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.

Synergistic effect of epigenetic inhibitors decitabine and suberoylanilide hydroxamic acid on colorectal cancer in vitro

Colorectal cancer (CRC) is major public health burden and a common cause of oncological deaths worldwide. Alterations of the epigenetic landscape are a well‐documented hallmark of CRC phenotype. Accumulation of aberrant DNA methylation and histone acetylation orchestrates to modify gene activity and drive tumor onset, progression and metastasis. In this study we evaluated the effect of suberoylanilide hydroxamic acid (SAHA), a pan histone deacetylase inhibitor, and decitabine (DAC), a DNA methyltransferase inhibitor, either alone or in combination, on Caco‐2 human colon cancer cell line in vitro. Our results showed that SAHA and DAC, separately, significantly decreased cell proliferation, induced apoptosis and cell cycle arrest of Caco‐2 cell line. On the other hand, the sequential treatment of Caco‐2 cells, first with DAC and then with SAHA, induced a synergistic anti‐tumor effect; the sequential treatment significantly enhanced growth inhibition and apoptosis of Caco‐2 cell line in comparison to cells treated with either drug alone. Furthermore, the combination therapy upregulates protein expression levels of pro‐apoptotic proteins Bax, p53 and cytochrome c, down regulates the expression of anti‐apoptotic Bcl‐2 protein and increases the cleavage of procaspases 8 and 9; this suggests that the combination activates apoptosis via both the intrinsic and extrinsic pathways. Mechanistically, we demonstrated that the synergistic anti‐neoplastic activity of combined SAHA and DAC involves an effect on PI3K/AKT and Wnt/β‐catenin signaling pathways as it down regulates p‐PI3K, p‐AKT, Wnt1, p‐GSK3, cyclin A1+A2, cyclin E1 and up regulates p‐β‐catenin protein expression levels. In conclusion, our results provide evidence for the profound anti‐tumorigenic effect of sequentially combined SAHA and DAC in CRC cell line and offer new insights into the corresponding underlined molecular mechanism.Support or Funding InformationThis work was funded by SRDC, School of Arts and Sciences, Lebanese American UniversityThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Enhanced Solubility, Permeability and Anticancer Activity of Vorinostat Using Tailored Mesoporous Silica Nanoparticles.

Suberoylanilide hydroxamic acid (SAHA) or vorinostat (VOR) is a potent inhibitor of class I histone deacetylases (HDACs) that is approved for the treatment of cutaneous T-cell lymphoma. However, it has the intrinsic limitations of low water solubility and low permeability which reduces its clinical potential especially when given orally. Packaging of drugs within ordered mesoporous silica nanoparticles (MSNs) is an emerging strategy for increasing drug solubility and permeability of BCS (Biopharmaceutical Classification System) class II and IV drugs. In this study, we encapsulated vorinostat within MSNs modified with different functional groups, and assessed its solubility, permeability and anti-cancer efficacy in vitro. Compared to free drug, the solubility of vorinostat was enhanced 2.6-fold upon encapsulation in pristine MSNs (MCM-41-VOR). Solubility was further enhanced when MSNs were modified with silanes having amino (3.9 fold) or phosphonate (4.3 fold) terminal functional groups. Moreover, permeability of vorinostat into Caco-2 human colon cancer cells was significantly enhanced for MSN-based formulations, particularly MSNs modified with amino functional group (MCM-41-NH2-VOR) where it was enhanced ~4 fold. Compared to free drug, vorinostat encapsulated within amino-modified MSNs robustly induced histone hyperacetylation and expression of established histone deacetylase inhibitor (HDACi)-target genes, and induced extensive apoptosis in HCT116 colon cancer cells. Similar effects were observed on apoptosis induction in HH cutaneous T-cell lymphoma cells. Thus, encapsulation of the BCS class IV molecule vorinostat within MSNs represents an effective strategy for improving its solubility, permeability and anti-tumour activity.

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.

Computer-aided drug design and virtual screening of targeted combinatorial libraries of mixed-ligand transition metal complexes of 2-butanone thiosemicarbazone

The present paper deals with in silico evaluation of 32 virtually designed transition metal complexes of 2-butanone thiosemicarbazone and N,S,O containing donor hetero-ligands namely py, bpy, furan, thiophene, 2-picoline, 1,10-phenanthroline, piperazine and liquid ammonia. The complexes were designed with a view to assess their potential anticancer, antioxidant and antibacterial activity. The absorption, distribution, metabolism, excretion and toxicity (ADMET) properties of the chosen ligands were calculated by admetSAR software. Metabolic sites of different ligands likely to undergo metabolism were predicted using Metaprint 2D. The proposed complexes were also evaluated for their drug-like quality based on Lipinski’s, Veber, Ghose and leadlikeness filters. Druglikeness and toxicity potential were predicted by OSIRIS property explorer. The pharmacokinetic properties and bioactivity scores were calculated by Molinspiration tool. Bioactivity scores of the complexes were predicted for drug targets including enzymes, nuclear receptors, kinase inhibitors, G-protein coupled receptor ligands and ion channel modulators. Molecular docking of selected Fe(II) mixed-ligand complexes was performed using AutoDock version 4.2.6 and i-GEMDOCK version 2.1 with two target proteins namely Ribonucleotide reductase (RR) and Topoisomerase II (Topo II). The results were compared with three standard reference drugs viz. Doxorubicin HCl, Letrozole (anticancer) and Tetracycline (antibiotic). Multivariate data obtained were analyzed using principal component analysis (PCA) for visualization and projection as scatter and 3D plots. Positive results obtained for hetero-ligands using admetSAR version 1.0 indicated good absorption and transport kinetics of the hetero-ligand complexes through the human intestine and blood-brain barrier. The hetero-ligands were predicted to have no associated mutagenic effect(s) also. However, none of the hetero-ligands was predicted to be Caco-2 (human colon cancer cell line) permeable. Most of the hetero-ligands and the parent ligand (2-butanone thiosemicarbazone) were predicted to undergo Phase-I metabolism prior to excretion using MetaPrint2D software. Pharmacokinetic evaluation of the proposed complexes revealed that all complexes displayed drug-like character and were predicted to have no apparent toxic side-effects. All the proposed complexes showed moderate to good biological activity scores (−5.0 to 0.0). Mixed complexes with bpy, 2-picoline and 1,10-phenanthroline showed significant bioactivity scores (as enzyme inhibitors) in the range 0.02–0.13. Likewise, good docking scores were obtained for Fe (II) complexes with the same ligands. [FeL(2-pic)2] displayed the lowest binding energy (−6.47 kcal/mol) with respect to Topo II followed by [FeL(py)2] (−6.14 kcal/mol) as calculated by AutoDock version 4.2.6. With respect to binding with RR, [FeL(2--pic)2] again displayed the lowest binding energy (−7.21 kcal/mol) followed by [FeL(py)2] (−5.96 kcal/mol). On the basis of docking predictions and various other computational evaluations, four mixed-ligand complexes of Fe in +2 oxidation state with py, bpy, 2--picoline and 1,10-phenanthroline were synthesized with 2-butanone thiosemicarbazone. All the synthesized Fe complexes were characterized using various spectroscopic techniques and tested for their potential anticancer activity in vitro against human breast carcinoma cell line MDA-MB 231 and human lung carcinoma cell line A549 cell line using MTT assay. [FeL(2-pic)2], [FeL(bpy)], and [FeL(py)2] were found to exhibit significant antiproliferative activity with IC50 values in the range of 80–100 μM against breast and lung cancer cells. The synthesized Fe complexes also displayed mild antioxidant activities. The synthesized and studied Fe complexes have the potential for development into future anticancer agents if analyzed and modified further for improvement of their ADMET, solubility and permeability criteria set for potential drug-candidates.

Chlorogenic Acid and Its Microbial Metabolites Exert Anti-Proliferative Effects, S-Phase Cell-Cycle Arrest and Apoptosis in Human Colon Cancer Caco-2 Cells.

Chlorogenic acid (CGA) decreases colon cancer-cell proliferation but the combined anti-cancer effects of CGA with its major colonic microbial metabolites, caffeic acid (CA), 3-phenylpropionic acid (3-PPA) and benzoic acid (BA), needs elucidation as they occur together in colonic digesta. Caco-2 cancer cells were treated for 24 h with the four compounds individually (50–1000 µM) and as an equimolar ratio (1:1:1:1; MIX). The effective concentration to decrease cell proliferation by 50% (EC50) was lower for MIX (431 ± 51.84 µM) and CA (460 ± 21.88) versus CGA (758 ± 19.09 µM). The EC50 for cytotoxicity measured by lactate dehydrogenase release in MIX (527 ± 75.34 µM) showed more potency than CA (740 ± 38.68 µM). Cell proliferation was decreased by 3-PPA and BA at 1000 µM with no cytotoxicity. Cell-cycle arrest was induced at the S-phase by CA (100 µM), MIX (100 µM), CGA (250 µM) and 3-PPA (500 µM) with activation of caspase-3 by CGA, CA, MIX (500 and 1000 µM). Mitochondrial DNA content was reduced by 3-PPA (1000 µM). The anti-cancer effects occurred at markedly lower concentrations of each compound within MIX than when provided singly, indicating that they function together to enhance anti-colon cancer activities.

Short Chain Fatty Acids Enhance Aryl Hydrocarbon (Ah) Responsiveness in Mouse Colonocytes and Caco-2 Human Colon Cancer Cells

Aryl hydrocarbon receptor (AhR) ligands are important for gastrointestinal health and play a role in gut inflammation and the induction of T regulatory cells, and the short chain fatty acids (SCFAs) butyrate, propionate and acetate also induce similar protective responses. Initial studies with butyrate demonstrated that this compound significantly increased expression of Ah-responsive genes such as Cyp1a1/CYP1A1 in YAMC mouse colonocytes and Caco-2 human colon cancer cell lines. Butyrate synergistically enhanced AhR ligand-induced Cyp1a1/CYP1A1 in these cells with comparable enhancement being observed for 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and also microbiota-derived AhR ligands tryptamine, indole and 1,4-dihydroxy-2-naphthoic acid (DHNA). The effects of butyrate on enhancing induction of Cyp1b1/CYP1B1, AhR repressor (Ahrr/AhRR) and TCDD-inducible poly(ADP-ribose)polymerase (Tiparp/TiPARP) by AhR ligands were gene- and cell context-dependent with the Caco-2 cells being the most responsive cell line. Like butyrate and propionate, the prototypical hydroxyamic acid-derived histone deacetylase (HDAC) inhibitors Panobinostat and Vorinostat also enhanced AhR ligand-mediated induction and this was accompanied by enhanced histone acetylation. Acetate also enhanced basal and ligand-inducible Ah responsiveness and histone acetylation, demonstrating that acetate was an HDAC inhibitor. These results demonstrate SCFA-AhR ligand interactions in YAMC and Caco-2 cells where SCFAs synergistically enhance basal and ligand-induced expression of AhR-responsive genes.