HT-29 Colon Adenocarcinoma Cells Research Articles

Effects of Size Polydispersity and Dense Media on Quantitative Ultrasound Estimates.

Quantitative ultrasound (QUS) techniques based on the backscatter coefficient (BSC) aim to characterize the scattering properties of biological tissues. A scattering model is fit to the measured BSC, and the fitted QUS parameters can provide local tissue microstructure, namely, scatterer size and acoustic concentration. However, these techniques may fail to provide a correct description of tissue microstructure when the medium is polydisperse and/or dense. The objective of this study is to investigate the effects of scatterer size polydispersity in sparse or dense media on the QUS estimates. Four scattering models (i.e., the monodisperse and polydisperse sparse models, and the monodisperse and polydisperse concentrated models based on the structure factor) are compared to assess their accuracy and reliability in quantifying the QUS estimates. Simulations are conducted with different scatterer size distributions for sparse, moderately dense, and dense media (volume fractions of 1%, 20%, and 73%, respectively). The QUS parameters are estimated by using model-based inverse methods at different center frequencies between 8 and 50 MHz. Experimental data are also analyzed using colon adenocarcinoma HT29 cell pellet biophantoms to further validate the results obtained from simulations at the volume fraction of 73%. Our findings reveal that the choice of scattering model has a significant impact on the accuracy of QUS estimates. For sufficiently high frequencies and dense media, the polydisperse concentrated model outperforms the other models and enables more accurate quantification. Furthermore, our results contribute to advancing our understanding of the complexities associated with scatterer size polydispersity and dense media in spectral-based QUS techniques.

Doxorubicin-induced chemoresistance in Duke's type B colon adenocarcinoma cell line is aggravated in the presence of TGF-β2 through non-apoptotic cell death.

The current challenge in clinical cancer treatment is chemoresistance. Colon cells have inherently higher xenobiotic transporters expression and hence can attain resistance rapidly. Increased levels of TGF-β2 expression in patients have been attributed to cancer progression, aggressiveness, and resistance. To investigate resistance progression, we treated doxorubicin (dox) to HT-29 colon adenocarcinoma cells in the presence or absence of TGF-β2 ligand. After 1, 3-, and 7-day treatment, we investigated cell proliferation, viability, and cytotoxicity by MTT, trypan blue staining, and lactate dehydrogenase enzyme release. The mechanism of cell death was elucidated by hoechst33342 and propidium iodide dual staining and apoptosis assay. The development of resistance was detected by rhodamine123 efflux and P-glycoprotein (P-gp)/MDR1 antibody staining through fluorimetry and flow cytometry. The colony formation ability of the cells was also elucidated. Inhibition of cell proliferation was noted after day 1, while a significant reduction in viability and a significant increase in lactate dehydrogenase release was detected after day 3. Reduction of intracellular rhodamine123 levels was detected after day 3 and was significantly lower in dox with TGF-β2 treatment compared to dox alone. Increased surface P-gp levels after days 3 and 7 were observed in the treated groups. Hoechst33342/propidium iodide staining and apoptosis assay indicated non-apoptotic cell death. The cells treated with TGF-β2 had higher colony formation ability. TGF-β2 expression might play a significant role in the development of chemoresistance to doxorubicin in Duke's type B colon adenocarcinoma cell line, HT-29.

Evaluation of the Antitumor Activity of Quaternary Ammonium Surfactants.

Quaternary ammonium surfactants, due to their diverse chemical structure and their biological properties, can be used in medicine as DNA carriers, disinfectants, and antimicrobial and antitumor agents. In this study, using melanoma A375, colon adenocarcinoma HT-29 and normal human dermal fibroblast (NHDF) cells, we tested the hypothesis that the quaternary ammonium surfactants 2-dodecanoyloxyethyl)trimethylammonium bromide (DMM-11), 2-dodecanoyloxypropyl)trimethylammonium bromide (DMPM-11) and 2-pentadecanoyloxymethyl)trimethylammonium bromide (DMGM-14) act selectively against cancer cells. The results showed that these compounds led to the initiation of the apoptotic process of programmed cell death, as evidenced by the ratio of the relative expression of Bax protein to Bcl-2. The encapsulation of surfactants in liposomes allowed lower concentrations to be used. Moreover, encapsulation reduced their toxicity towards non-cancerous cells. The anticancer efficiency and apoptotic effect of the liposomal formulations with surfactants (DMM-11, DMPM-11 and DMGM-14) were higher than those of surfactant-free liposomes. Therefore, quaternary ammonium surfactant-loaded liposomes show significant potential as delivery vehicles for the treatment of melanoma and colon cancers. The use of nano-formulations offers the advantage of optimizing quaternary ammonium surfactant delivery for improved anticancer therapy.

Evaluation of the Effect of Betulin and Its Alkynyl Derivatives on the Profile of Changes in Gene Expression of the Inflammatory Process of Colorectal Adenocarcinoma Cells (HT-29 Cell Line)

Betulin is a lupane-type pentacyclic triterpene. It is characterized by a range of biological properties, including anti-cancer and anti-inflammatory activities. It is also an origin compound for obtaining derivatives with higher biological activity and better bioavailability. Chronic inflammation stimulates the formation of a pro-cancer microenvironment, promoting tumor growth, cell migration, and neoangiogenesis. Many factors, immune system cells, and cytokines and chemokines released by them are involved in this process. Therefore, it has been suggested that the optimal target for anti-cancer drugs in this disease could be substances showing anti-inflammatory activity. The aim of the study was to indicate the direction of changes in the expression of genes related to the inflammatory state in colorectal cancer cells promoted by betulin and its selected alkynyl derivatives. Cytotoxicity assessment was carried out using a sulforhodamine B (SRB) test, whereas lipophilicity was determined by reversed-phase thin-layer chromatography (RP-TLC). The analysis of the gene expression profile in colon adenocarcinoma cells treated with betulin and its derivatives was performed using oligonucleotide microarrays HG-U133A. Based on the conducted analysis, it can be stated that betulin and its derivatives 1–3 influence the change in the expression profile of genes related to inflammatory processes in the HT-29 colon adenocarcinoma cell lines. The highest expression changes (FC > 2) were observed for HMOX1 (compound 1 vs. control) and TMED7 (compound 3 vs. control) mRNAs. An important observation is the comparison of the profile of changes in the expression of the studied genes in the compared compounds. Derivative 1 showed the greatest similarity to control cells, whereas betulin showed similarity to cisplatin. These observations indicate the necessity further research on the impact of betulin and its derivatives on inflammatory processes and the possible direction of chemical modification of compounds.

Evaluation of Cytotoxic Activity of Loaded Catechin into Iron Oxide Nanoparticles Coated with Sodium Alginate and Hydroxyapatite against Human HT-29 Colon Adenocarcinoma and Breast Cancer MCF-7 Cells

Evaluation of Cytotoxic Activity of Loaded Catechin into Iron Oxide Nanoparticles Coated with Sodium Alginate and Hydroxyapatite against Human HT-29 Colon Adenocarcinoma and Breast Cancer MCF-7 Cells

Synthesis and Biological Evaluation of Novel Benzylidene Thiazolo Pyrimidin-3(5H)-One Derivatives

Starting compound 1 was synthesized according to reference. 1 Benzylidene thiazole pyrimidin-3(5H)-ones were synthesized reactions of 1 with bromoacetic acid and various aryl-aldehydes in the same vessel via one-step, unlike studies in the literature. Quantum chemical parameters and full geometry optimizations for all compounds were computed using DFT based on B3LYP. Cytotoxic action potential of synthesized compounds was evaluated using trypan blue dye exclusion and MTT assays in different cell lines including adenocarcinoma alveolar basal epithelial-like adherent A549 cells, the colon adenocarcinoma HT-29 cells, prostate adenocarcinoma DU-145 cells, and diploid ARPE-19 retinal pigment epithelial cells. Embryotoxicity and genotoxicity assessments were performed on pluripotent human embryonal carcinoma NT2 and human lymphocyte cells, respectively. Compound A1 exhibited good anticancer activity on A549 and DU-145 cell lines, and the compounds including A3, 4, 6, and 9 induced cytotoxicity on A549 cells. The compounds A1-10 also showed a good biosafety profile at relatively lower concentrations.

Carborane-Based Tebufelone Analogs and Their Biological Evaluation In Vitro.

The presence of inflammatory mediators in the tumor microenvironment, such as cytokines, growth factors or eicosanoids, indicate cancer-related inflammatory processes. Targeting these inflammatory mediators and related signal pathways may offer a rational strategy for the treatment of cancer. This study focuses on the incorporation of metabolically stable, sterically demanding, and hydrophobic dicarba-closo-dodecaboranes (carboranes) into dual cyclooxygenase-2 (COX-2)/5-lipoxygenase (5-LO) inhibitors that are key enzymes in the biosynthesis of eicosanoids. The di-tert-butylphenol derivative tebufelone represents a selective dual COX-2/5-LO inhibitor. The incorporation of meta- or para-carborane into the tebufelone scaffold resulted in eight carborane-based tebufelone analogs that show no COX inhibition but 5-LO inhibitory activity in vitro. Cell viability studies on HT29 colon adenocarcinoma cells revealed that the observed antiproliferative effect of the para-carborane analogs of tebufelone is enhanced by structural modifications that include chain elongation in combination with introduction of a methylene spacer resulting in higher anticancer activity compared to tebufelone. Hence, this strategy proved to be a promising approach to design potent 5-LO inhibitors with potential application as cytostatic agents.

Beyond Antioxidant Activity: Redox Properties of Catechins May Affect Changes in the DNA Methylation Profile-The Example of SRXN1 Gene.

The role of catechins in the epigenetic regulation of gene expression has been widely studied; however, if and how this phenomenon relates to the redox properties of these polyphenols remains unknown. Our earlier study demonstrated that exposure of the human colon adenocarcinoma HT29 cell line to these antioxidants affects the expression of redox-related genes. In particular, treatment with (-)-epigallocatechin (EGC) downregulated transcription of gene encoding sulfiredoxin-1 (SRXN1), the peroxidase involved in the protection of cells against hydrogen peroxide-induced oxidative stress. The aim of this study was to investigate whether the observed SRXN1 downregulation was accompanied by changes in the DNA methylation level of its promoter and, if so, whether it was correlated with the redox properties of catechins. The impact on DNA methylation profile in HT29 cells treated with different concentrations of five catechins, varying in chemical structures and standard reduction potentials as well as susceptibility to oxidation, was monitored by a methylation-sensitive high-resolution melting technique employing the SRXN1 promoter region as a model target. We demonstrated that catechins, indeed, are able to modulate DNA methylation of the SRXN1 gene in a redox-related manner. The nonlinear method in the statistical analysis made it possible to fish out two parameters (charge transfer in oxidation process Qox and time of electron transfer t), whose strong interactions correlated with observed modulation of DNA methylation by catechins. Based on these findings, we present a proof-of-concept that DNA methylation, which limits SRXN1 expression and thus restricts the multidirectional antioxidant action of SRXN1, may represent a mechanism protecting cells against reductive stress caused by particularly fast-reacting reductants such as EGC and (-)-epicatechin gallate (ECG) in our study.

Phenolic Compounds from Sour Cherry Pomace: Microencapsulation, in Vitro Digestion, and Cell Growth Activities.

The objective of this work was the valorisation of sour cherry (Prunus cerasus L.) pomace as a source of biologically active compounds. To formulate microcapsules, polyphenolic compounds were extracted and encapsulated with maltodextrin as wall material, by freeze-drying. An in vitro digestion study was carried out on obtained encapsulates but also on sour cherry pomace extract and sour cherry pomace freeze-dried powder. The results indicated that encapsulation, as well as freeze-drying, provided a good protective effect on bioactive compounds during digestion. Furthermore, the potential antiproliferative and cytotoxic activities of encapsulates and sour cherry pomace extract were evaluated using breast adenocarcinoma MCF7 cell lines, colon adenocarcinoma HT-29 cell lines, and noncancer cell line. Encapsulates and sour cherry pomace extract showed variable anti-proliferative activity towards all cell lines. Obtained results showed that encapsulation of sour cherry pomace could be useful for improving the stability of polyphenolic compounds in the gastrointestinal tract. The results highlight the bioactive potential of sour cherry pomace as a nutraceutical resource and the protective effects of microencapsulation on the digestion of bioactive compounds.

Native Size-Exclusion Chromatography–Based Mass Spectrometry Reveals New Components of the Early Heat Shock Protein 90 Inhibition Response Among Limited Global Changes

The molecular chaperone heat shock protein 90 (HSP90) works in concert with co-chaperones to stabilize its client proteins, which include multiple drivers of oncogenesis and malignant progression. Pharmacologic inhibitors of HSP90 have been observed to exert a wide range of effects on the proteome, including depletion of client proteins, induction of heat shock proteins, dissociation of co-chaperones from HSP90, disruption of client protein signaling networks, and recruitment of the protein ubiquitylation and degradation machinery—suggesting widespread remodeling of cellular protein complexes. However, proteomics studies to date have focused on inhibitor-induced changes in total protein levels, often overlooking protein complex alterations. Here, we use size-exclusion chromatography in combination with mass spectrometry (SEC-MS) to characterize the early changes in native protein complexes following treatment with the HSP90 inhibitor tanespimycin (17-AAG) for 8 h in the HT29 colon adenocarcinoma cell line. After confirming the signature cellular response to HSP90 inhibition (e.g., induction of heat shock proteins, decreased total levels of client proteins), we were surprised to find only modest perturbations to the global distribution of protein elution profiles in inhibitor-treated HT29 cells at this relatively early time-point. Similarly, co-chaperones that co-eluted with HSP90 displayed no clear difference between control and treated conditions. However, two distinct analysis strategies identified multiple inhibitor-induced changes, including known and unknown components of the HSP90-dependent proteome. We validate two of these—the actin-binding protein Anillin and the mitochondrial isocitrate dehydrogenase 3 complex—as novel HSP90 inhibitor-modulated proteins. We present this dataset as a resource for the HSP90, proteostasis, and cancer communities (https://www.bioinformatics.babraham.ac.uk/shiny/HSP90/SEC-MS/), laying the groundwork for future mechanistic and therapeutic studies related to HSP90 pharmacology. Data are available via ProteomeXchange with identifier PXD033459.

Influence of different food models and storage temperatures on the bacterial growth inhibition by maltodextrin laurate and sucrose laurate and investigation of their cytotoxicity

Maltodextrin esterified to laurate (ML) was synthesized enzymatically and compared to commercial sucrose laurate (SL) for microbial growth-inhibitory properties in food systems. There was broad microbial inhibition by ML in growth media, but practical information about various foods and storage temperatures is limited with no toxicological information. This research investigated the antimicrobial property of ML against two spoilage bacteria (Bacillus subtilis and Lactobacillus plantarum) in growth media and related food models stored at different temperatures (4 °C, 22 °C, and 35 °C). Additionally, in vitro cytotoxicity of ML was investigated in human colon adenocarcinoma HT-29 cells and human normal colon epithelial CCD 841 CoN cells using a standard colorimetric assay. ML was highly inhibitive against L. plantarum in apple juice at all temperatures tested. SL could inhibit both bacteria in the food models tested (milk, apple juice, tomato ketchup, French dressing) at 4 °C. ML was not observed to be cytotoxic to either cell line up to 500 μg/mL, whereas SL was evidently cytotoxic with IC50 ranging from 84 to 94 μg/mL. In conclusion, ML can be an alternative to commercial SL as a safe food additive in certain products.

Investigation of the potential antitumor activity of PLK1 inhibitor SBE13 in colon cancer cell line HT29

Background: High levels of Polo-like kinase 1 (PLK1), which have been found to be abnormally expressed in many tumor types, are known to contribute to tumorigenesis and poor prognosis. Therefore, specific targeting of PLK1 is an important strategy for cancer therapy. In this study, it was aimed to investigate the cytotoxic effect of SBE13, one of the PLK1 inhibitors, against HT29 colon adenocarcinoma cells and its apoptotic potential.
 Methods: The cytotoxic effect of SBE13 on HT29 was determined by XTT colorimetric assay. Flow cytometry was also used to determine apoptosis. 
 Results: SBE13 showed a dose-dependent cytotoxic effect in HT29 cells and its IC50 value was calculated as 11.79 µM for 48 h. Moreover, the Annexin V binding assay revealed that SBE13 treatment significantly increased the apoptosis in HT29 cells.
 Conclusion: Generally, SBE13 exerts a cytotoxic effect promoted by apoptosis in colon cancer cells HT29. Although the anticancer efficacy of SBE13 in colon cancer is promising, this potential effect should be reinforced by further studies.

Regulation of apoptosis and autophagy by albendazole in human colon adenocarcinoma cells

Albendazole (ABZ) was initially introduced as an anthelmintic, however, many studies have reported with its anticancer effects. We investigated the anti-tumor effects of ABZ in vitro in human colon adenocarcinoma HCT-15, HCT-116, HT-29, and SW480 cell lines in this study. The cytotoxicity of ABZ was analyzed in colon adenocarcinoma cell lines and normal CCD18Co cells. We found that ABZ induced the subG1 arrest during cell cycle progression, increased the late apoptotic cells, shifted of peak TUNEL-labeled cells peak, and induced apoptosis. Then effects on autophagy activation was confirmed by acridine orange (AO), MDC staining, and immunocytochemistry of LC3. It was observed that ABZ can induce the autophagy activation through modulating the levels of LC3, Atg7, and beclin-1. For mechanistic studies, apoptosis blocker (Z-DEVD-FMK) and autophagy inhibitor (3-MA) were used to confirm that whether ABZ has apoptosis and autophagy specific effects, and reversal in both these cell death processes were noted. The effects of ABZ on AMPK, MAPKs, and ULK induction was also evaluated. We noticed that N-acetyl cysteine (NAC), a broad spectrum antioxidant, can effectively inhibit both apoptosis and autophagy. However, ABZ could even recover suppression of apoptosis and autophagy caused by NAC in colon cancer cells. Therefore, ABZ can potentially up-regulate both the apoptosis and autophagy to significantly suppress tumorigenesis in colorectal cancer cell lines.

α-Bisabolol Mitigates Colon Inflammation by Stimulating Colon PPAR-γ Transcription Factor: In Vivo and In Vitro Study.

The incidence and prevalence of inflammatory bowel disease (IBD, Crohn's disease, and ulcerative colitis) are increasing worldwide. The etiology of IBD is multifactorial, including genetic predisposition, dysregulated immune response, microbial dysbiosis, and environmental factors. However, many of the existing therapies are associated with marked side effects. Therefore, the development of new drugs for IBD treatment is an important area of investigation. Here, we investigated the anti-inflammatory effects of α-bisabolol, a naturally occurring monocyclic sesquiterpene alcohol present in many aromatic plants, in colonic inflammation. To address this, we used molecular docking and dynamic studies to understand how α-bisabolol interacts with PPAR-γ, which is highly expressed in the colonic epithelium: in vivo (mice) and in vitro (RAW264.7 macrophages and HT-29 colonic adenocarcinoma cells) models. The molecular docking and dynamic analysis revealed that α-bisabolol interacts with PPAR-γ, a nuclear receptor protein that is highly expressed in the colon epithelium. Treatment with α-bisabolol in DSS-administered mice significantly reduced Disease Activity Index (DAI), myeloperoxidase (MPO) activity, and colonic length and protected the microarchitecture of the colon. α-Bisabolol treatment also reduced the expression of proinflammatory cytokines (IL-6, IL1β, TNF-α, and IL-17A) at the protein and mRNA levels. The expression of COX-2 and iNOS inflammatory mediators were reduced along with tissue nitrite levels. Furthermore, α-bisabolol decreased the phosphorylation of activated mitogen-activated protein kinase (MAPK) signaling and nuclear factor kappa B (NFκB) proteins and enhanced colon epithelial PPAR-γ transcription factor expression. However, the PPAR-α and β/δ expression was not altered, indicating α-bisabolol is a specific stimulator of PPAR-γ. α-Bisabolol also increased the PPAR-γ transcription factor expression but not PPAR-α and β/δ in pretreated in LPS-stimulated RAW264.7 macrophages. α-Bisabolol significantly decreased the expression of proinflammatory chemokines (CXCL-1 and IL-8) mRNA in HT-29 cells treated with TNF-α and HT-29 PPAR-γ promoter activity. These results demonstrate that α-bisabolol mitigates colonic inflammation by inhibiting MAPK signaling and stimulating PPAR-γ expression.

Structure-performance relationships of four lysosomal markers used for the imaging of HT-29 cancer cells and a cellular model of lysosomal storage disease (Niemann-Pick C)

Four new BODIPY derivatives with a potential tendency to aggregation have been synthesized and characterized by means of NMR techniques, mass spectrometry and UV–Vis/fluorescence spectroscopies. The objective of this study is to determine which structural factors of the new molecules influence most notably the cellular uptake, intracellular location and fluorescence imaging abilities. The behaviour of the compounds in organic solvent and aqueous solution has been studied. In organic solvents (DMSO, ethanol and toluene), the photophysical properties of the new molecules are almost independent of the building blocks used to synthesize the pendant moieties (non-fluorogenic parts). In an aqueous environment (HEPES Buffer Solution, pH 7), at 10 μM, three of the compounds (1, 2 and 4) tend to form weakly emissive nanoparticles (DLS determination) whereas one of them (3) remains soluble and highly fluorescent. In the nanomolar range of concentration, all the compounds are aqueous soluble. The cellular internalisation of the compounds (10 nM) has been studied in human colon adenocarcinoma HT-29 cells by means of flow cytometry and confocal laser scanning microscopy. All the compounds were uptaken by HT-29 cells, but notably molecule 3 (made with lysine as a building block) was the one displaying a higher loading and a more clear lysosomal location (0.88 Pearson's correlation coefficient in colocalization assays using lysosomal fluorescent probe LysoTracker DND-99). Molecule 3 also performed better than the valine derivative 1 as a lysosomal marker in a cellular model (human adrenal carcinoma SW13/cl.2 cells) of lysosomal storage disease (Niemann-Pick type C).

Synthesis, physicochemical characterization, and in vitro evaluation of biodegradable PLGA nanoparticles entrapped to folic acid for targeted delivery of kaempferitrin.

Polymeric nanoparticles are widely studied in the treatment of colorectal cancer. Kaempferitrin-loaded nontoxic and biodegradable poly(d,l-lactic-co-glycolic acid) (PLGA) nanoparticles (NPs) developed by the solvent emulsion evaporation method by improving its solubility and bioavailability. In order to improve the delivery of kaempferitrin (KM) to cancerous cells, folic acid (FA) combined kaempfertrin PLGA NPs were prepared. The goal of the study was whether PLGA NPs with surface KM and FA could help to prevent colorectal cancer. The synthesis of KM with FA in a nanomedicine could be crucial in the development of colon cancer chemotherapeutics. The physicochemical characteristics of synthesized KM-entrapped PLGA NPs were investigated by XRD, FTIR, zeta potential, and TEM. The KM + FA + PLGA NPs showed particle size with 132.9 ± 1.4 nm, zeta potential -15.0 ± 1.73 mV, encapsulation efficiency 67.92 ± 4.8, and drug-loading capacity 0.463 ± 0.173. In vitro cytotoxicity study on HT-29 cell lines using the MTT assay, the apoptotic study revealed that KM + FA + PLGA NPs have an enhanced cytotoxic effect compared to the KM + PLGA NPs drug solution. These findings suggested that KM + FA + PLGA NPs could be an effective chemotherapeutic drug delivery system in colon adenocarcinoma HT-29 cells.

Combination of resveratrol and BIBR1532 inhibits proliferation of colon cancer cells by repressing expression of LncRNAs.

Colorectal cancer (CRC) is the third most common cancer worldwide. The development of tumor drug resistance is observed in the treatment of CRC. Combinations of anticancer agents are attracting considerable interest in order to overcome drug resistance in CRC. This study aims to investigate the effect of resveratrol and BIBR1532, either alone or in combination, on the cell viability as well as on expression of long non-coding RNAs (LncRNAs) for HT-29 colon adenocarcinoma cells. The cytotoxic effects of resveratrol and BIBR1532 on HT-29 cells were determined using WST-1 test. Flow cytometry was used to determine apoptotic cell death after treatments. Real-Time PCR was used to identify expression of LncRNAs after treatments. LncExpDB and GEPIA2 were used to evaluate expression profiles of LncRNAs, whose expression levels were decreased in HT-29 cells after treatments, in normal tissues and colon adenocarcinoma tumors. IC50 concentrations of BIBR1532 and resveratrol were found to be 50.81μM at 48h and 86.23μM at 72h, respectively. Combination index value was 1.07617. BIBR1532, resveratrol, or their combination reduced the cell viability of HT-29 cells. CCAT1, CRNDE, HOTAIR, PCAT1, PVT1, SNHG16 were down-regulated after treatments. In silico analysis revealed that LncRNAs whose expression levels were decreased after treatments were associated with CRC. Resveratrol, BIBR1532, or their combination may have anti-proliferative effect on colorectal cancer cells through repressing expression of LncRNAs that are involved in progression of CRC.

A comparative study of supercritical fluid and ethanol extracts of cannabis inflorescences: Chemical profile and biological activity

We compared chemical profiles and in vitro biological activity of extracts obtained through maceration with ethanol or supercritical fluid extraction with CO2 (scCO2), from two different cannabis inflorescences chemotypes, one with high total tetrahydrocannabinol (THC) and other with high total cannabidiol (CBD) content. scCO2 was performed to obtain extracts with highest total THC and CBD recovery using ethanol as a co-solvent. In addition, a short maceration with ethanol was performed for both chemotypes. Cannabinoid and terpene composition of the extracts were determined using chromatographic and spectroscopic methods, and their cytotoxicity and cell proliferation effects were evaluated in vitro using HT-29 colon adenocarcinoma cell line. In vitro biological activity confirmed that cannabis extracts from both chemotypes have anti-proliferative activity but the scCO2 extract from high total THC chemotype was more cytotoxic. These results underscore the importance of chemotype and extraction method selection in the preparation of cannabis extracts for different therapeutic applications.

Development of enteric-coated, biphasic chitosan/HPMC microcapsules for colon-targeted delivery of anticancer drug-loaded nanoparticles

Oral delivery of anticancer drug-loaded nanoparticles (NPs) to the colon offers opportunities to improve colorectal cancer (CRC) treatment by increasing the free drug concentration at tumour sites and/or enhancing NP accumulation in tumours. Indomethacin, 5-FU and curcumin, were entrapped separately in Eudragit RS NPs (approximately 10% w/w loading) using nanoprecipitation and incorporated in biphasic chitosan/HPMC microcapsules (MCs) using aerosolisation. The MCs were designed to release NPs primarily in the colon following chitosan breakdown by bacterial enzymes. Around 10% of the drug-loaded NPs was released from MCs in simulated intestinal fluid (SIF) in 6 h and 20% in simulated colon fluid (SCF). Indomethacin release from MCs was absent in simulated gastric fluid (SGF) and restricted to around 10% in SIF and SCF, respectively, demonstrating potential for delivering a large fraction of contained drug to the colon. Curcumin release from NPs or NP-loaded MCs was negligible in SGF, SIF and SCF, revealing opportunities for delivery of curcumin-loaded NPs to the colon for accumulation in tumours. Curcumin-loaded NPs reduced proliferation of human colon adenocarcinoma HT-29 cells by 83% compared with 50% for free curcumin. These findings demonstrate the potential of chitosan/HPMC microcapsules as a colon-specific delivery vehicle for oral nanomedicines directed against colorectal cancer.

Decreased Levels of Microfibril-Associated Glycoprotein (MAGP)-1 in Patients with Colon Cancer and Obesity Are Associated with Changes in Extracellular Matrix Remodelling

Objective: The protein microfibril-associated glycoprotein (MAGP)-1 constitutes a crucial extracellular matrix protein. We aimed to determine its impact on visceral adipose tissue (VAT) remodelling during obesity-associated colon cancer (CC). Methods: Samples obtained from 79 subjects (29 normoponderal (NP) (17 with CC) and 50 patients with obesity (OB) (19 with CC)) were used in the study. Circulating concentrations of MAGP-1 and its gene expression levels (MFAP2) in VAT were analysed. The impact of inflammation-related factors and adipocyte-conditioned media (ACM) on MFAP2 mRNA levels in colon adenocarcinoma HT-29 cells were further analysed. The effects of MAGP-1 in the expression of genes involved in the extracellular matrix (ECM) remodelling and tumorigenesis in HT-29 cells was also explored. Results: Obesity (p < 0.01) and CC (p < 0.001) significantly decreased MFAP2 gene expression levels in VAT whereas an opposite trend in TGFB1 mRNA levels was observed. Increased mRNA levels of MFAP2 after the stimulation of HT-29 cells with lipopolysaccharide (LPS) (p < 0.01) and interleukin (IL)-4 (p < 0.01) together with a downregulation (p < 0.05) after hypoxia mimicked by CoCl2 treatment was observed. MAGP-1 treatment significantly enhanced the mRNA levels of the ECM-remodelling genes collagen type 6 α3 chain (COL6A3) (p < 0.05), decorin (DCN) (p < 0.01), osteopontin (SPP1) (p < 0.05) and TGFB1 (p < 0.05). Furthermore, MAGP-1 significantly reduced (p < 0.05) the gene expression levels of prostaglandin-endoperoxide synthase 2 (COX2/PTGS2), a key gene controlling cell proliferation, growth and adhesion in CC. Interestingly, a significant decrease (p < 0.01) in the mRNA levels of MFAP2 in HT-29 cells preincubated with ACM from volunteers with obesity compared with control media was observed. Conclusion: The decreased levels of MAGP-1 in patients with obesity and CC together with its capacity to modulate key genes involved in ECM remodelling and tumorigenesis suggest MAGP-1 as a link between AT excess and obesity-associated CC development.