Human Colon Adenocarcinoma Cell Line Research Articles

Sorafenib acts synergistically in combination with radiotherapy without causing intestinal damage in colorectal cancer.

Colorectal cancer is one of the commonest cancers. Chemoradiotherapy gives better results than radiotherapy or chemotherapy in colorectal cancer. To enhance radiosensitivity of tumor cells for chemoradiotherapy, targeted therapy drugs that act as radiosensitizers can be used. In the present study, we provide a scientific rationale for the clinical application of sorafenib as a radiosensitizer in colorectal cancer, without causing significant adverse effects on normal intestinal tissue. Three human colorectal adenocarcinoma cell lines (HCT116, HT-29, and SW480) were treated with sorafenib alone, or radiation followed by sorafenib. In vitro tests were performed using colony forming assays, cell cycle analysis, and comet assays. In addition, the effects of sorafenib and radiation therapy on the inhibition HT-29 tumor growth and survival of intestinal jejunum crypts were examined in vivo. Sorafenib increased the radiosensitivity of tumor cells in human colon adenocarcinoma cell lines (HCT116, HT-29, and SW480), as well as in HT-29 xenograft animal models. Sorafenib, in combination with ionizing radiation, induced the accumulation of tumor cells in the G2-M phase and delayed the repair of DNA damage caused by ionizing radiation. The combination of sorafenib and ionizing radiation did not enhance the apoptosis of intestinal crypt cells, compared with the use of radiation alone. We provide a scientific rationale for the use of sorafenib in combination with radiotherapy in colorectal cancer.

The Oxime Derivatives of 1-R-1H-Naphtho[2,3-d][1,2,3]triazole-4,9-dione 2-oxides: Synthesis and Properties.

To synthesize a novel chemotype based on the naphthoquinone scaffold with retained cytotoxicity and provisionally low intracellular oxidation potential. Derivatives of naphthoquinone, although potent anticancer agents, can exert heart toxicity due to generation of free oxygen species. In this study, we modified the scaffold by replacing one carbonyl group with the oxime moiety. Interestingly, only one carbonyl group in 1-R-1H-naphtho[2,3-d][1,2,3]triazole-4,9-dione 2-oxides reacted with hydroxylamine. The spatial structure was determined by X-ray analysis. New compounds were tested for the ability to form stable complexes with double stranded DNA by spectroscopy and molecular docking and to induce death of tumor cell lines and non-malignant counterparts. The resulting 1-R-1H-naphtho[2,3-d][1,2,3]triazole-4,9-dione 4-oxime 2-oxides were further acylated to produce a series of 1-R-1H-naphtho[2,3-d][1,2,3]triazole-4,9-dione 4-(O-acyloxime) 2-oxides. Newly synthesized compounds demonstrated a higher (in submicromolar or low micromolar range) cytotoxic potency against human colon and breast adenocarcinoma cell lines than to non-malignant skin fibroblasts. Spectroscopic measurements revealed that, unlike other classes of quinone derivatives, new naphthotriazoledione oxides did not form stable complexes with double stranded DNA regardless of their fitting to the DNA minor groove (as determined by molecular modeling). Thus, our chemical modifications yielded a new chemotype with good cytotoxic properties and yet-to-be-identified intracellular target(s).

Tissue Engineering Laboratory Models of the Small Intestine.

In recent years, three-dimensional (3D) cell culture models of the small intestine have gained much attention. These models support cell proliferation, migration, and differentiation and encourage tissue organization which is not possible in two-dimensional (2D) culture systems. Furthermore, the use of a wide variety of cell culture scaffolds and support substrates has revealed considerable differences in cell behavior and tissue organization. These systems have been used in combination with intestinal stem cells, organoid units, or human colonic adenocarcinoma cell lines such as Caco-2 and HT29-MTX to generate a number of in vitro and in vivo models of the intestine. In this study, we review the current 2D and 3D tissue engineering models of the intestine to determine the most effective sources of intestinal cells and current research on support scaffolds capable of inducing the morphological architecture and function of the intestinal mucosa.

Cytotoxic alkaloids against human colon adenocarcinoma cell line (HT-29) from the seed embryos of Nelumbo nucifera

Phytochemical investigation of the CHCl3 extract of the seed embryos of Nelumbo nucifera Gaertn resulted in the isolation of a new naturally occurring bisbenzylisoquinoline alkaloid, O-methylneferine (1), together with five known alkaloids, neferine (2), armepavine (3), (–)-(1R)-N-methylcoclaurine (4), nuciferine (5), and pronuciferine (6). The structures of these compounds were characterized by spectroscopic methods and comparison of physical properties with those reported in the literature. Among them, compounds 1 and 2 exhibited significant activity against human colon adenocarcinoma cell line (HT-29), with IC50 values of 0.70 and 1.61 µM, respectively, which were 8- and 3.5-fold higher than that of the reference anticancer drug, doxorubicin (IC50 5.63 µM). Moreover, compounds 1 and 2 displayed less cytotoxic activity against the non-cancerous HEK 239 cells with the IC50 values of 42.48 and 12.19 μM, respectively, whereas the cytotoxicity of doxorubicin against this cell line was 0.22 μM. The very potent cytotoxicity against HT-29 cell line and very high selectivity index (60.6-fold) of the alkaloid 1 is of particular significant; it could be considered as a promising structure lead for anti-colon cancer drug development.

Isolates of Lactobacillus plantarum and L. reuteri display greater antiproliferative and antipathogenic activity than other Lactobacillus isolates.

Lactic acid bacteria (LAB) have been associated with many beneficial effects in human digestive physiology. The aim of this study was to evaluate such effect, including attachment, antiproliferation and anti-pathogenic/antibacterial/antimicrobial properties of LAB isolated from healthy humans. Thirteen isolates, obtained from fecal samples of healthy individuals, were identified by phenotypic and molecular methods. Human colon adenocarcinoma cell line HT-29 and the cell proliferation kit II (XTT) assay were used for examination of the Lactobacillus adherence and antiproliferative activity, respectively. In addition, the inhibitory effect of Lactobacillus isolates against pathogenic bacteria was examined. Out of 13 Lactobacillus isolates, 5 (38 %) isolates were non-adhesive, 4 (31 %) were adhesive and 4 (31 %) were strongly adhesive. Amongst the isolated lactobacilli, L. reuteri showed the highest degree of inhibitory effect against the attachment of the enteropathogens. The XTT assay showed that 3 different isolates had the strongest antiproliferative activity with the maximum effect observed by L. plantarum isolates. Our results described that different Lactobacillus species isolated from normal fecal samples had different degrees of antiproliferative and anti-pathogenic/antibacterial/antimicrobial activities. However, no isolates showed all of the examined properties concurrently, suggestive that a combination of Lactobacillus species is needed for an active biological defense system.

64Cu-Labeled Repebody Molecules for Imaging of Epidermal Growth Factor Receptor-Expressing Tumors.

The epidermal growth factor receptor (EGFR) is a member of the erbB family of receptors and is overexpressed in many tumor types. A repebody is a newly designed nonantibody protein scaffold for tumor targeting that contains leucine-rich repeat modules. In this study, 3 64Cu-labeled anti-EGFR repebodies with different chelators were synthesized, and their biologic characteristics were assessed in cultured cells and tumor-bearing mice. Methods: Repebodies were synthesized with the chelators 2-(p-isothiocyanatobenzyl)-1,4,7-triazacyclononane-N,N',N,″-triacetic acid trihydrochloride ([p-SCN-Bn]-NOTA), 2,2',2″-(10-(2-(2,5-dioxopyrrolidin-1-yloxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl) triacetic acid (DOTA-N-hydroxysuccinimide ester), or 1-(p-isothiocyanatobenzyl)diethylenetriamine pentaacetic acid trihydrochloride ([p-SCN-Bn]-DTPA) in 1.0 M NaHCO3 buffer (pH 9.2) for 24 h. Purified NOTA-, DOTA-, and DTPA-conjugated repebody were radiolabeled with 64Cu in 0.1 M NH4OAc buffer (pH 5.5). To compare the EGFR-binding affinities of the repebodies, cellular uptake studies were performed with the human non-small cell lung cancer cell line H1650 (high expression of EGFR) and the human colon adenocarcinoma cell line SW620 (low expression of EGFR). Biodistribution and small-animal PET imaging studies were performed using H1650 tumor-bearing mice. Results: Radiochemical yields of the 64Cu-labeled repebodies were approximately 70%-80%. Cellular uptake of the NOTA-, DOTA-, and DTPA-repebodies was over 4-fold higher in H1650 cells than in SW620 cells at 1 h. The 3 repebodies had accumulated specifically in H1650 tumor-bearing nude mice by 1 h after intravenous injection and were retained for over 24 h, as measured by the percentage injected dose per gram of tissue (%ID/g). Tumor uptake of all repebodies increased from 1 to 6 h (at 1 h, 6.28, 8.46, and 6.91 %ID/g for NOTA-, DOTA-, and DTPA-repebody, respectively; at 6 h, 9.4, 8.28, and 10.1 %ID/g, respectively). H1650 tumors were clearly visible after injection of each repebody, with high tumor-to-background ratios (at 1 h, 3.43, 4.89, and 2.38 for NOTA-, DOTA-, and DTPA-repebody, respectively; at 6 h, 3.05, 4.36, and 2.08; at 24 h, 3.81, 4.58, and 2.86). Conclusion: The 3 64Cu-repebody complexes demonstrated specific and rapid uptake in EGFR-expressing tumors within 1 h and may have potential as novel EGFR imaging agents for PET.

Phytochemical Constituents from the Root of Luvunga Scandens and Biological Activity Evaluation

Isobutylglucosinolate (1) was isolated from the root of Luvunga scandens (Roxb.) for the first time, which demonstrated an antioxidant activity with an IC50 value of 30.1±0.2 μg/mL Additionally, two biological active phytosterols, named β-sitosterol (2) and stigmasterol (3) were also isolated from Luvunga scandens (Roxb.), showing the low cytotoxicity against human colon adenocarcinoma cell line (HT29) with IC50 values of 124±1 μg/mL and 141±3 μg/mL, respectively. The chemical structure of all isolated compounds (1–4) was identified by spectroscopic analyses.

Differential MicroRNA Expression Profile of Colon Cancer Stem Cells Derived from Primary Tumor and HT-29 Cell line

BackgroundColorectal cancer (CRC) is one of the most prevalent cancers in the world. Cancer stem cells (CSC) have been reported in many human tumors and are thought to be responsible for tumor initiation, therapy resistance, progression, relapse, and metastasis. Recent studies revealed that microRNAs (miRNA) play important roles in maintaining stemness of embryonic stem cells and CSCs.ObjectivesWith regard to the crucial role of cancer stem cells in colon cancer initiation and maintenance, the miRNA expression profile in primary and cell line cancer stem cells compared to non-stem cells cancer cells were evaluated.MethodsIn this study, a population of colon CSCs from primary colon cancer and human HT-29 colonic adenocarcinoma cell line was isolated from serum free medium and their microRNA profiles were evaluated using miRNA PCR array.ResultsThe isolated cells with high expression of EPCAMﰠ markers showed greater colony-forming efficiency and higher tumorigenic potential. Furthermore, expression of ‘‘stemness’’ genes including C-myc, Klf4, Nanog, Sox2 and Oct4 was higher in isolated cancer stem cells. Moreover, miRNA expression profile of colon CSCs was performed using miRNA PCR array. There are 39 differentially expressed miRNAs, 24 of which had lower mean expression in the cancer stem cells samples. By contrast, 15 miRNAs had higher expression levels in CSCs samples. Of these, miR-495, miR-125, and miR-199a were the most significantly up-regulated miRNAs.ConclusionsOur results suggest that miRNAs might play important roles in maintenance and regulation of colon CSCs and specific miRNA expression signatures may contribute to cancer initiation and expansion.

Studies on the cytotoxicity and anticancer performance of heterocyclic hypervalent organobismuth(III) compounds

Novel organobismuth(III) complex of 5H-dibenzo[c,f][1,5]oxabismocin-12(7H)-yl nitrate (C2) was synthesized and characterized by spectral and elemental analysis. It was compared with other five C,E,C-chelating (E = N, O, S) organobismuth(III) complexes against human adenocarcinoma alveolar basal epithelial cells (A549), human liver cancer cell line (SMCC7721), human gastric cancer cell line (SGC-7901), human colon adenocarcinoma cell line (SW480) and healthy human bronchial cell line (16HBE14o-) in vitro. It was found that C2 exhibited the best anticancer activity. Further mechanistic investigation indicated that toxicological activity of C2 was ascribable to apoptosis rather than anti-proliferative activity. Apoptosis was induced through up-regulating the level of Bcl-2/Bax as well as the activation of caspase-3. The results demonstrate that heterocyclic organobismuth(III) complexes of this type have great potential in the treatment of cancer.

Upregulation of Yes-associated protein and transcriptional co-activator with PDZ-binding motif influences the behavior of LOVO human colon adenocarcinoma cells.

The present study aimed to investigate the role of Yes-associated protein (YAP) and transcriptional co-activator with PDZ-binding motif (TAZ) in the LOVO human colon adenocarcinoma cell line and explore the underlying mechanisms. First, the expression levels of YAP and TAZ were detected in LOVO cells using reverse-transcription quantitative PCR, and the results suggested that YAP and TAZ were faintly expressed in LOVO cells. To investigate the exact role of YAP and TAZ in LOVO cells, stable YAP- and/or TAZ-overexpressing LOVO cell lines were established using YAP and/or TAZ expression plasmids. An MTT assay and flow cytometry were used to assess cell proliferation and apoptosis, respectively. The results indicated that compared with the control, YAP or TAZ overexpression significantly increased the proliferation ability of LOVO cells, while apoptosis was significantly decreased. Furthermore, the expression of the tumor-associated proteins connective tissue growth factor and cysteine-rich angiogenic inducer 61, which have critical roles in facilitating cancer cell proliferation, migration and invasion, were found to be upregulated following upregulation of YAP and TAZ. In addition, the expression of cell apoptosis-associated protein B-cell lymphoma 2 (Bcl-2) was significantly increased, while Bcl-2-associated X protein and caspase-3 were inhibited by YAP or TAZ overexpression. All of these effects were amplified when YAP and TAZ were co-overexpressed. In conclusion, YAP and TAZ function as tumor promoters in human colon carcinoma, and upregulation of YAP and TAZ influences the behavior of LOVO colon adenocarcinoma cells via regulating tumor-associated gene expression.

In vitro cytotoxic and antioxidant activities of Pikut Trichinthalamaga remedy

The Pikut Trichinthalamaga remedy appears in ancient Thai pharmacy scripture, as a purgative, and includes Rhubarb (Rheum palmatum L.), Myrabulan fruit (Terminalia chebula Retz.), and Gamboge (Garcinia handuryi Hook F.). We investigated the cytotoxic properties of the herbal extracts using the SRB assay in human colon adenocarcinoma cell lines (LS174T and SW480) and normal lung fibroblast cell (MRC-5) as well as their antioxidant activities by chemical-(ABTS assay) and cell-based assays [nitric oxide (NO) assay]. The active fractions were identified using high-performance liquid chromatography coupled to electrospray ionisation and quadruple time-of-flight mass spectrometry (HPLC-ESI-QTOF-MS). The ethanolic extract of Pikut Trichinthalamaga remedy had a cytotoxic effect on LS174T and SW 480 cells (IC50 = 0.39 and 0.51 μg/ml, respectively). The ABTS assay showed that the water extract of the myrabulan fruit had the highest activity (EC50 = 5.69 μg/ml) while the NO assay showed that the ethanolic extract of Pikut Trichinthalamaga remedy had the most potent inhibitory activity on NO production (EC50 = 0.62 μg/ml). After liquid–liquid partitioning sequence, the chloroform fraction had the highest cytotoxicity. Emodin, forbesione, gaudichaudionic acid and gaudichaudionol were identified in this extract. These results support the further investigation of the Pikut Trichinthalamaga remedy as a potential anticancer drug.

NGR-peptide-drug conjugates with dual targeting properties.

Peptides containing the asparagine-glycine-arginine (NGR) motif are recognized by CD13/aminopeptidase N (APN) receptor isoforms that are selectively overexpressed in tumor neovasculature. Spontaneous decomposition of NGR peptides can result in isoAsp derivatives, which are recognized by RGD-binding integrins that are essential for tumor metastasis. Peptides binding to CD13 and RGD-binding integrins provide tumor-homing, which can be exploited for dual targeted delivery of anticancer drugs. We synthesized small cyclic NGR peptide–daunomycin conjugates using NGR peptides of varying stability (c[KNGRE]-NH2, Ac-c[CNGRC]-NH2 and the thioether bond containing c[CH2-CO-NGRC]-NH2, c[CH2-CO-KNGRC]-NH2). The cytotoxic effect of the novel cyclic NGR peptide-Dau conjugates were examined in vitro on CD13 positive HT-1080 (human fibrosarcoma) and CD13 negative HT-29 (human colon adenocarcinoma) cell lines. Our results confirm the influence of structure on the antitumor activity and dual acting properties of the conjugates. Attachment of the drug through an enzyme-labile spacer to the C-terminus of cyclic NGR peptide resulted in higher antitumor activity on both CD13 positive and negative cells as compared to the branching versions.

Validation of the cell line LS180 as a model for study of the gastrointestinal toxicity of mycophenolic acid

1. Gastrointestinal (GI) intolerability is a concern for drugs such as mycophenolic acid (MPA) and drug metabolism may play a role. Few in vitro models exist that allow for the preclinical evaluation of a potential role of drug metabolism in intestinal drug toxicity. Thus, we sought to develop an in vitro model based on the human colon adenocarcinoma cell line LS180 to investigate MPA’s negative effects on intestinal cells.2. Stability of expression of key enzymes of MPA metabolism (UGT1A7, UGT1A9, UGT1A10, UGT2B7, CYP3A4 and CYP3A5), transporters (OATP1B1, OATP1B3, OATP2B1, MRP1, MRP2 and MDR1) and the nuclear receptor PXR over 12 passages in combination with guanosine supplementation to counter MPA’s antiproliferative effects (determined by western blot analysis and proliferation assays, respectively) was established.3. Expression of LS180 key enzymes remained stable over passages 47–59 and MPA-induced growth inhibition was circumvented by exogenous guanosine over a period of three days. MPA was not cytotoxic at concentrations up to 250 μM, a concentration that intestinal cells adjacent to the dissolving capsule or tablet are exposed to.4. We concluded that LS180 cells are suitable to study the potential association between MPA metabolism and its negative effects on intestinal cells.

Evaluation of anticancer activity of water and juice extracts of young Hordeum vulgare in human cancer cell lines HT-29 and A549.

Introduction and objective. Barley (Hordeum vulgare L.) is known as a rich source of different bioactive compounds. At present, considerable attention of researchers is focused on young barley grass. It can be a good source of dietary minerals, vitamins, carbohydrates, amino acids, phenolic compounds and proteins. It is possible that the composition of chemical ingredients beneficial for health may induce an anticancer potential of young barley in human colon adenocarcinoma (HT-29) and human lung adenocarcinoma (A549) cell lines. Materials and method. Hordeum vulgare water extract (HWE) and Hordeum vulgare juice extract (HJE) were prepared. Cell proliferation and viability were examined with the use of MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) and NR (neutral red) methods. Induction of necrosis was assessed by propidium iodide/Hoechst staining. Progress of the cell cycle involved in cell proliferation, apoptosis, and regulation of transcription was estimated using flow cytometry analysis. Additionally, the capability of free radical scavenging was evaluated with the DPPH assay. Results. The study revealed that extracts inhibited the proliferation of cancer cells. The NR study confirmed the low cytotoxic activity of the tested extracts to normal human colon epithelial cells (CCD 841 CoTr) and human skin fibroblasts (HSF). Furthermore, a dose-dependent cytotoxicity against HT-29 cells, but not A549 cells, has been reported. The free radical scavenging activity was observed in the case of the HWE but not the HJE. Conclusions. The obtained results indicate a cancer chemopreventive potential of young barley as a safe dietary agent in colon carcinoma.

Twist1 confers multidrug resistance in colon cancer through upregulation of ATP-binding cassette transporters.

Multidrug resistance is a major problem in colon cancer treatment. However, its molecular mechanisms remain unclear. Recently, the epithelial-mesenchymal transition (EMT) in anticancer drug resistance has attracted increasing attention. This study investigated whether vincristine treatment induces EMT and promotes multidrug resistance in colon cancer. The result showed that vincristine treatment increases the expression of several ATP-binding cassette transporters in invasive human colon adenocarcinoma cell line (HCT-8). Vincristine-resistant HCT-8 cells (HCT-8/V) acquire a mesenchymal phenotype, and thus its migratory and invasive ability are increased both in vitro and in vivo. The master transcriptional factors of EMT, especially Twist1, were significantly increased in the HCT-8/V cell line. Moreover, the ectopic expression of Twist1 increased the chemoresistance of HCT-8 cells to vincristine and increased the expression levels and promoter activities of ABCB1 and ABCC1. Furthermore, Twist1 silencing reverses the EMT phenotype, enhances the chemosensitivity of HCT-8/ V cells to anticancer agents in vitro and in vivo, and downregulates the expression of ABCB1 and ABCC1. Twist1-mediated promotion of ABCB1 and ABCC1 expression levels plays an important role in the drug resistance of colon cancer cells.

SGLT1-Mediated Transport in Caco-2 Cells Is Highly Dependent on Cell Bank Origin

The human colon adenocarcinoma (Caco-2) cell line is a well-established in vitro model for studying transport phenomena for prediction of intestinal nutrient and drug absorption. However, substances depending on transporters such predictions are complicated due to variable transporter expression and limited knowledge about transporter function during multiple cell passaging and cell thawings. In the case of sodium glucose transporter 1 (SGLT1), a key transporter of oral absorption of d-glucose, one reason for compromised prediction could be inadequate expression of SGLT1 in Caco-2 cells and thereby limited sensitivity in the determination of SGLT1-mediated permeability (PSGLT1). Here, the objective is to characterize and compare SGLT1-mediated uptake in Caco-2 cells obtained from different cell banks. SGLT1-mediated uptake of the standard SGLT1 substrate, methyl-α-d-glucopyranoside, in Caco-2 cells was shown to be highly dependent on cell bank origin. The most robust and reliable SGLT1 functionality was identified in Caco-2 cells from Deutsche Sammlung für Mikroorganismen und Zellkulturen (DSMZ), whereas cells from the American Type Culture Collection and European Collection of Authenticated Cell Cultures have lower SGLT1 transport activity. Transepithelial PSGLT1 across Caco-2 cells from DSMZ showed that PSGLT1 likely accounts for approximately 97% of absorptive methyl-α-d-glucopyranoside Papp(a-b). In conclusion, Caco-2 cells from DSMZ provide a robust in vitro model for studying SGLT1-mediated uptake and transport—over multiple cell passages and independent cell stock thawings.

Zeta-potential-changing nanoparticles conjugated with cell-penetrating peptides for enhanced transfection efficiency.

The aim of this study was to develop zeta-potential-changing nanoparticles (NPs) combining cell-penetrating peptides for gene delivery. NPs were formed using phosphorylated carboxymethyl cellulose-glucosamine 6-phosphate (CMC-G6P) and polyethylene imine-polyarginine conjugates. Phosphate release was evaluated using intestinal alkaline phosphatase and cell lines. Transfection studies with plasmid DNA were then performed. The zeta potential of CMC-G6P/branched PEI NPs was -3 mV and switched to +4 mV after intestinal alkaline phosphatase cleavage. The released phosphate in human colon adenocarcinoma cell line was more pronounced than human embryonic kidney cell line 293. Transfection studies demonstrated the greatest expression of plasmid DNA when being incorporated into CMC-G6P/polyethylene imine-polyarginine NPs. Novel zeta potential changing NPs combining cell-penetrating peptides are a promising tool to deliver DNA drugs to target cells.

Evaluation of Cytotoxicity of Aqueous Extract of <I>Euphorbia hirta</I> Using Human Colon Adenocarcinoma and Vero Cell Line

Euphorbia hirta Linn. belonging to family Euphorbiaceae . Traditionally it is used in the treatment of tumors, fever, asthma, bronchitis, chest congestion, etc. Objective of the present investigation was to evaluate cytotoxicity of aqueous extract of E. hirta using human colon adenocarcinoma (HCT-15) and vero cell line. MTT assay was used to evaluate cytotoxicity of aqueous extract of E. hirta using HCT-15 and vero cell line. Doxorubicin was used as standard reference drug. The concentrations of extract and doxorubicin at range 0.05-1000 μg/ml were considered. Results of the present study was revealed that the 50% cell growth inhibition (IC50) of aqueous extract of E. hirta and doxorubicin was 510.66 μg/ml and 460.13μg/ml using the HCT-15 cell line and 1363.65 μg/ml and 2392.71μg/ml using Vero cell line respectively. The cytotoxic effect of aqueous extract of E. hirta may be due to the presence of previously reported chemical compounds by earlier scientists in the plant. This plant can be used as good source of cytotoxic compounds. Chemical analysis of this extract is in under pipeline. First time author have reported these studies on E. hirta using HCT-15 and vero cell line by MTT assay. Further series of studies will required for validating these results by using in-vivo pharmacological experiments. The work can be further expanded for detail mechanistic based studies by doing Tunnal, Caspcase and flow cytometry assays.

Variability of antioxidant and biological activities of Rhus tripartitum related to phenolic compounds.

Rhus species are known in traditional medicine for their therapeutic virtue and their extracts showed numerous important properties including antimalarial, antimicrobial, antiviral, and hypoglycemic and anticonvulsant activities. Rhus tripartitum (Ucria) is a medicinal plant widely used in Tunisia folk medicine against chronic diarrhea and gastric ulcer. This study was designed to examine in vitro and ex vivo antioxidant, anti-inflammatory and anticancer activities of four extracts of Rhus tripartitum root cortex with increasing solvent polarity (hexane, dichloromethane, methanol and water). HPLC was used to identify and quantify phenolic compounds in Rhus extract. Water extract showed the highest antioxidant activity using oxygen radical absorbance capacity (ORAC method) with 8.95 ± 0.47 µmol Trolox/mg and a cell based-assay with 0.28 ± 0.12 µmol Trolox/mg as compared to the other fractions. Moreover, methanol extract displayed the strongest anti-cancer activity against human lung carcinoma (A-549) and colon adenocarcinoma cell lines (DLD-1) with an IC50 value of 60.69 ± 2.58 and 39.83 ± 4.56 µg/ml (resazurin test) and 44.52 ± 5.96 and 55.65 ± 6.00 µg/ml (hoechst test), respectively. Besides, the highest anti-inflammatory activity, inhibiting nitric oxide (NO) release, was exhibited by dichloromethane extract with 31.5 % at 160 µg/ml in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. The HPLC analysis showed that catechol and kaempferol were the major phenolics. These data suggest the richness of all fractions of Ucria root on interesting bioactive molecules with different polarity and confirm the known traditional therapeutics virtues of this species for the treatment of dysentery, diarrhea and gastric ulcer.

Transferrin-targeted poly(butylene adipate)/terephthalate nanoparticles for targeted delivery of 5-fluorouracil in HT29 colorectal cancer cell line

The purpose of this study was to design 5-fluorouracil-loaded poly(butylene adipate)/terephthalate (Ecoflex®) nanoparticles for targeting colorectal cancer. The nanoparticles were prepared by emulsification–solvent evaporation method and optimized by a full factorial design. The effects of polymer and surfactant concentration, surfactant type, and stirrer rate were studied on the particle size, zeta potential, loading efficiency, and release efficiency of nanoparticles. For production of targeted nanoparticles, chitosan was conjugated to transferrin which was then coated on the surface of Ecoflex nanoparticles via electrostatic interactions. The conjugation of transferrin/chitosan was verified by Fourier transform infrared spectroscopy, ultraviolet spectroscopy, and SDS-PAGE (sodium dodecyl sulfate–polyacrylamide gel electrophoresis) methods and quantified by ultraviolet spectroscopy assay. The cytotoxicity of 5-fluorouracil loaded in targeted and non-targeted nanoparticles was studied on human colon adenocarcinoma cell line (HT29), Michigan Cancer Foundation-7 (MCF-7), and human umbilical vein endothelial cells using MTT (thiazolyl blue tetrazolium bromide) assay. The best results were obtained from nanoparticles prepared by 0.2% of the polymer, 2% of Tween 20, and stirrer speed of 17,500 r/min. The successful conjugation of transferrin/chitosan was confirmed by Fourier transform infrared spectrum and SDS-PAGE results and was about 80%. The targeted nanoparticles showed significantly more cytotoxic effects on HT29 cells compared to free 5-fluorouracil and non-targeted nanoparticles. Blocking transferrin receptors resulted in a significantly higher cell survival for targeted nanoparticles which confirmed receptor-mediated cellular uptake of targeted nanoparticles.