Phyllanthus emblica L. Fruit Extract Induces Chromosomal Instability and Suppresses Necrosis in Human Colon Cancer Cells

Phyllanthus emblica L. (PE) is an edible fruit indigenous to Southeast Asia. It has been considered as a potent functional food due to its numerous pharmacological applications, such as anti-oxidant, antimicrobial, anti-diabetic and protection for multiple organs. The aim of this study was to evaluate the effects of a water extract of PE fruit on genomic damage and cell death in the human colon adenocarcinoma cell line COLO320 using the cytokinesis-block micronucleus cytome assay. Cells were exposed to RPMI-1640 medium containing 0, 20, 40, 80, or 160 μg/mL PE for 24, 48, 72, or 96 hours. The results showed that PE induced a significant decrease in necrosis (p < 0.001) and nuclear division index (NDI) (p < 0.001) in a dose- and time-dependent manner, and there was a highly significant correlation between the reduction of necrosis and NDI (r = 0.820, p < 0.001). Dose- and time-dependent increases (p < 0.001) in the frequency of chromosomal instability (CIN) were observed after PE exposure, and the frequency of CIN was negatively correlated with NDI (r = - 0.640, p < 0.001). PE also significantly increased apoptosis (p < 0.001), and there was a significant correlation of apoptosis with CIN (r = 0.566, p < 0.001). In conclusion, PE suppresses necrosis and delays mitotic progression, which results in massive CIN followed by apoptosis in COLO320 cells.

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Safed Musli (Chlorophytum borivilianum L.) Callus-Mediated Biosynthesis of Silver Nanoparticles and Evaluation of their Antimicrobial Activity and Cytotoxicity against Human Colon Cancer Cells

Journal of Nanomaterials ◽

10.1155/2019/2418785 ◽

2019 ◽

Vol 2019 ◽

pp. 1-8 ◽

Cited By ~ 4

Author(s):

Fengchang Huang ◽

Yaxin Long ◽

Qingqing Liang ◽

Boregowda Purushotham ◽

Mallappa Kumara Swamy ◽

...

Keyword(s):

Biomedical Applications ◽

Colon Adenocarcinoma ◽

Human Colon ◽

Dependent Manner ◽

Face Centered Cubic ◽

Human Colon Cancer ◽

Chlorophytum Borivilianum ◽

The Face ◽

Average Size ◽

Safed Musli

With the advancement of nanobiotechnology, eco-friendly approaches of plant-mediated silver nanomaterial (AgNP) biosynthesis have become more attractive for biomedical applications. The present study is a report of biosynthesizing AgNPs using Chlorophytum borivilianum L. (Safed musli) callus extract as a novel source of reducing agent. AgNO3 solution challenged with the methanolic callus extract displayed a change in color from yellow to brown owing to the bioreduction reaction. Further, AgNPs were characterized by using UV–visible spectrophotometry, X-ray Diffraction (XRD), Atomic Force Microscopy (AFM), and Fourier Transform Infrared Spectroscopy (FTIR). UV–vis spectrum revealed the surface plasmon resonance property of AgNPs at around 450 nm. XRD pattern with typical peaks indicated the face-centered cubic nature of silver. AFM analysis confirmed the existence of spherical-shaped and well-dispersed AgNPs having an average size of 52.0 nm. Further, FTIR analysis confirmed the involvement of different phytoconstituents of the callus extract role in the process of bioreduction to form nanoparticles. The AgNPs were more efficient in inhibiting the tested pathogenic microbes, namely, Pseudomonas aeruginosa, Bacillus subtilis, Methicillin-resistant Escherichia coli, Staphylococcus aureus, and Candida albicans compared to callus extract. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay confirmed the cytotoxic property of AgNPs against human colon adenocarcinoma cell line (HT-29) in a dose-dependent manner. At higher concentrations of 500 μg/mL AgNPs, the cell viability was observed to be only 7% after 24 hours with IC50 value of 254 μg/mL. Therefore, these AgNPs clearly endorse the manifold potential to be used in various biomedical applications in the near future.

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In Vitro Antiproliferative and Antioxidant Effects of Extracts from Rubus caesius Leaves and Their Quality Evaluation

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2016/5698685 ◽

2016 ◽

Vol 2016 ◽

pp. 1-8 ◽

Cited By ~ 5

Author(s):

Daniel Mirosław Grochowski ◽

Roman Paduch ◽

Adrian Wiater ◽

Adrianna Dudek ◽

Małgorzata Pleszczyńska ◽

...

Keyword(s):

Ethyl Acetate ◽

Antioxidant Activities ◽

Water Extract ◽

Ethyl Acetate Fraction ◽

Human Colon ◽

Dependent Manner ◽

Human Colon Cancer ◽

Concentration Dependent Manner ◽

Two Stages

The present study was performed to evaluate the effect of different extracts and subfractions from Rubus caesius leaves on two human colon cancer cell lines obtained from two stages of the disease progression lines HT29 and SW948. Tested samples inhibited the viability of cells, both HT29 and SW948 lines, in a concentration-dependent manner. The most active was the ethyl acetate fraction which, applied at the highest concentration (250 μg/mL), decreased the viability of cells (HT29 and SW948) below 66%. The extracts and subfractions were also investigated for antioxidant activities on DPPH and FRAP assays. All extracts, with the exception of water extract at a dose of 250 μg/mL, almost totally reduced DPPH. The highest Fe3+ ion reduction was shown for the diethyl and ethyl acetate fractions. It was more than 6.5 times higher (at a dose 250 μg/mL) as compared to the control. The LC-MS studies of the analysed preparations showed that all samples contain a wide variety of polyphenolics, among which ellagitannins turned out to be the main constituents with dominant ellagic acid, sanguiin H-6, and flavonol derivatives.

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Reversal of Multidrug Resistance in Human Colon Cancer and Human Leukemia Cells by Three Plant Extracts and Their Major Secondary Metabolites

Medicines ◽

10.3390/medicines5040123 ◽

2018 ◽

Vol 5 (4) ◽

pp. 123 ◽

Cited By ~ 7

Author(s):

Jun-Xian Zhou ◽

Michael Wink

Keyword(s):

Colon Cancer ◽

Cell Line ◽

Abc Transporters ◽

Human Colon ◽

Rhodamine 123 ◽

Human Leukemia ◽

Dependent Manner ◽

Human Colon Cancer ◽

High Concentrations ◽

Resistant Cells

Background: We studied the effect of three plant extracts (Glycyrrhiza glabra, Paeonia lactiflora, Eriobotrya japonica) and six of their major secondary metabolites (glycyrrhizic acid, 18β glycyrrhetinic acid, liquiritigenin, isoliquiritigenin, paeoniflorin, ursolic acid) on the multidrug resistant human colon cancer cell line Caco-2 and human leukemia cell line CEM/ADR 5000 as compared to the corresponding sensitive cell line CCRF-CEM, and human colon cancer cells HCT-116, which do not over-express ATP-binding cassette (ABC) transporters. Methods: The cytotoxicity of single substances in sensitive and resistant cells was investigated by MTT assay. We also applied combinations of extracts or single compounds with the chemotherapeutic agent doxorubicin or doxorubicin plus the saponin digitonin. The intracellular retention of the ABC transporter substrates rhodamine 123 and calcein was examined by flow cytometry to explore the effect of the substances on the activity of ABC transporters P-glycoprotein and MRP1. Real-time PCR was applied to analyse the gene expression changes of ABCB1, ABCC1, caspase 3, caspase 8, AhR, CYP1A1, and GSTP1 in resistant cells under the treatment of the substances. Results: All the substances moderately inhibited cell growth in sensitive and resistant cells to some degree. Whereas ursolic acid showed IC50 of 14 and 22 µM in CEM/ADR 5000 and Caco-2 cells, respectively, glycyrrhizic acid and paeoniflorin were inactive with IC50 values above 400 μM. Except for liquiritigenin and isoliquiritigenin, all the other substances reversed MDR in CEM/ADR 5000 and Caco-2 cells to doxorubicin. Ue, ga, 18ga, and urs were powerful reversal agents. In CEM/ADR 5000 cells, high concentrations of all the substances, except Paeonia lactiflora extract, increased calcein or rhodamine 123 retention in a dose-dependent manner. In Caco-2 cells, all the substances, except liquiritigenin, retained rhodamine 123 in a dose-dependent manner. We also examined the effect of the plant secondary metabolite (PSM) panel on the expression of ABCB1, ABCC1, caspase 3, caspase 8, AhR, CYP1A1, and GSTP1 genes in MDR cells. Conclusions: The extracts and individual PSM could reverse MDR in CEM/ADR 5000 and Caco-2 cells, which overexpress ABC transporters, in two- and three-drug combinations. Most of the PSM also inhibited the activity of ABC transporters to some degree, albeit at high concentrations. Ue, ga, 18ga, and urs were identified as potential multidrug resistance (MDR) modulator candidates, which need to be characterized and validated in further studies.

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Phosphofructokinase 2 and glycolysis in HT29 human colon adenocarcinoma cell line. Regulation by insulin and phorbol esters

Biochemical Journal ◽

10.1042/bj2680465 ◽

1990 ◽

Vol 268 (2) ◽

pp. 465-470 ◽

Cited By ~ 10

Author(s):

C Denis-Pouxviel ◽

T Gauthier ◽

D Daviaud ◽

J C Murat

Keyword(s):

Phorbol Esters ◽

Lactate Production ◽

Colon Adenocarcinoma ◽

Glucose Consumption ◽

Human Colon ◽

Kinetic Properties ◽

Dependent Manner ◽

Glycerol Phosphate ◽

Ht29 Cells ◽

Long Term Treatment

Kinetic properties of phosphofructokinase 2 (PFK2) and regulation of glycolysis by phorbol 12-myristate 13-acetate (PMA) and insulin were investigated in highly glycolytic HT29 colon cancer cells. PFK2 was found to be inhibited by citrate and, to a lesser extent, by phosphoenolpyruvate and ADP, but to be insensitive to inhibition by sn-glycerol phosphate. From these kinetic data, PFK2 from HT29 cells appears different from the liver form, but resembles somewhat the heart isoenzyme. Fructose 2,6-bisphosphate (Fru-2,6-P2) levels, glucose consumption and lactate production are increased in a dose-dependent manner in HT29 cells treated with PMA or insulin. The increase in Fru-2,6-P2 can be related to an increase in the Vmax. of PFK2, persisting after the enzyme has been precipitated with poly(ethylene glycol), without change in the Km for fructose 6-phosphate. The most striking effects of PMA and insulin on Fru-2,6-P2 production are observed after long-term treatment (24 h) and are abolished by actinomycin, cycloheximide and puromycin, suggesting that protein synthesis is involved. Furthermore, the effects of insulin and PMA on glucose consumption, lactate production, Fru-2,6-P2 levels and PFK2 activity are additive, and the effect of insulin on Fru-2,6-P2 production is not altered by pre-treatment of the cells with the phorbol ester. This suggests that these effects are exerted by separate mechanisms.

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Potentially probiotic Lactobacillus strains with anti-proliferative activity induce cytokine/chemokine production and neutrophil recruitment in mice

Beneficial Microbes ◽

10.3920/bm2016.0202 ◽

2017 ◽

Vol 8 (4) ◽

pp. 615-623 ◽

Cited By ~ 4

Author(s):

G. Saxami ◽

A. Karapetsas ◽

P. Chondrou ◽

S. Vasiliadis ◽

E. Lamprianidou ◽

...

Keyword(s):

Human Colon ◽

Intercellular Adhesion Molecule ◽

Probiotic Properties ◽

Dependent Manner ◽

Colony Stimulating Factor ◽

Human Colon Cancer ◽

Chemokine Production ◽

Air Pouch ◽

Lactobacillus Pentosus ◽

Stimulating Factor

Lactobacillus pentosus B281 and Lactobacillus plantarum B282 are two Lactobacillus strains previously isolated from fermented table olives. Both strains were found to possess probiotic properties and displayed desirable technological characteristics for application as starters in novel functional food production. In the present study the anti-proliferative and immunostimulatory activities of the two strains were investigated. Firstly, we demonstrated that live L. pentosus B281 and L. plantarum B282 significantly inhibited the growth of human colon cancer cells (Caco-2) in a time- and dose-dependent manner. By employing the air pouch system in mice, we showed that administration of both strains led to a rapid and statistically significant infiltration of leukocytes in the air pouch exudates. The phenotypical characterisation of the recruited immune cells was performed by flow cytometry analysis. We demonstrated that the majority of the infiltrated leukocytes were neutrophils. Finally by using the Mouse Cytokine Array Panel A Detection Antibody cocktail, we showed that both strains induced the expression of granulocyte-colony stimulating factor, interleukin (IL)-1α, IL-1β, IL-6, chemokine (C-X-C motif) ligand (CXCL)-1, chemokine (C-C motif) ligand (CCL)-3, CCL-4, and CXCL-2 and diminished the expression levels of soluble intercellular adhesion molecule, macrophage colony-stimulating factor and metallopeptidase inhibitor 1. Our results showed that both strains display anti-proliferative and immunostimulatory properties equal or even better in some cases than those of established and commonly used probiotic strains. These findings further support the probiotic character of the two strains.

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Expression of IGF-II and IGF binding proteins in differentiating human intestinal Caco-2 cells

AJP Endocrinology and Metabolism ◽

10.1152/ajpendo.1995.269.5.e804 ◽

1995 ◽

Vol 269 (5) ◽

pp. E804-E813 ◽

Cited By ~ 5

Author(s):

Y. Zhang ◽

D. A. Wick ◽

B. Seetharam ◽

N. M. Dahms

Keyword(s):

Conditioned Medium ◽

Binding Proteins ◽

Colon Adenocarcinoma ◽

Human Colon ◽

Metabolic Effects ◽

Mrna Levels ◽

Dependent Manner ◽

Proliferating Cells ◽

Igf Binding Proteins ◽

Igf Binding

The mitogenic and metabolic effects of insulin-like growth factor-II (IGF-II) can be modulated by six distinct IGF binding proteins (IGFBPs). As a first step toward understanding the role of IGFs and their binding proteins in intestinal epithelial cell differentiation, the expression of IGF-II and IGFBPs was characterized in the human colon adenocarcinoma Caco-2 cell line. Northern blot analysis revealed two IGF-II transcripts of 5.4 and 4.5 kb, and ribonuclease protection assays indicated that IGF-II mRNA levels are regulated during Caco-2 differentiation. A specific radioimmunoassay detected IGF-II in serum-free conditioned medium, the level of which was three- to fivefold higher in proliferating cells than in differentiated cells. Immunoprecipitation and ligand blot analyses of conditioned medium demonstrated that IGFBP-2, IGFBP-3, IGFBP-4, and IGFBP-6 are synthesized by Caco-2 cells, with IGFBP-2 and IGFBP-4 being the major IGFBPs secreted, and that the levels of IGFBP-2 and IGFBP-6 decreased as differentiation proceeded. These results indicate that the expression of IGF-II, IGFBP-2, and IGFBP-6 is regulated in a differentiation-dependent manner in Caco-2 cells.

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The Interplay between Fe3O4 Superparamagnetic Nanoparticles, Sodium Butyrate, and Folic Acid for Intracellular Transport

International Journal of Molecular Sciences ◽

10.3390/ijms21228473 ◽

2020 ◽

Vol 21 (22) ◽

pp. 8473

Author(s):

Maria Teresa Cambria ◽

Giusy Villaggio ◽

Samuele Laudani ◽

Luca Pulvirenti ◽

Concetta Federico ◽

...

Keyword(s):

Folic Acid ◽

Sodium Butyrate ◽

Intracellular Transport ◽

Colon Adenocarcinoma ◽

Human Colon ◽

Superparamagnetic Nanoparticles ◽

High Dose ◽

Dependent Manner ◽

The Difference ◽

Surface Modified

Combined treatments which use nanoparticles and drugs could be a synergistic strategy for the treatment of a variety of cancers to overcome drug resistance, low efficacy, and high-dose-induced systemic toxicity. In this study, the effects on human colon adenocarcinoma cells of surface modified Fe3O4 magnetic nanoparticles (MNPs) in combination with sodium butyrate (NaBu), added as a free formulation, were examined demonstrating that the co-delivery produced a cytotoxic effect on malignant cells. Two different MNP coatings were investigated: a simple polyethylene glycol (PEG) layer and a mixed folic acid (FA) and PEG layer. Our results demonstrated that MNPs with FA (FA-PEG@MNPs) have a better cellular uptake than the ones without FA (PEG@MNPs), probably due to the presence of folate that acts as an activator of folate receptors (FRs) expression. However, in the presence of NaBu, the difference between the two types of MNPs was reduced. These similar behaviors for both MNPs likely occurred because of the differentiation induced by butyrate that increases the uptake of ferromagnetic nanoparticles. Moreover, we observed a strong decrease of cell viability in a NaBu dose-dependent manner. Taking into account these results, the cooperation of multifunctional MNPs with NaBu, taking into consideration the particular cancer-cell properties, can be a valuable tool for future cancer treatment.

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Antiproliferation and anti-migration induced by gypenosides in human colon cancer SW620 and esophageal cancer Eca-109 cells

Human & Experimental Toxicology ◽

10.1177/0960327113497771 ◽

2013 ◽

Vol 33 (5) ◽

pp. 522-533 ◽

Cited By ~ 20

Author(s):

H Yan ◽

X Wang ◽

Y Wang ◽

P Wang ◽

Y Xiao

Keyword(s):

Colon Cancer ◽

Flow Cytometry ◽

Esophageal Cancer ◽

Cell Viability ◽

Cancer Cells ◽

Human Colon ◽

Dependent Manner ◽

Colon Cancer Cells ◽

Human Colon Cancer ◽

Esophageal Cancer Cells

Gypenosides (Gyp), found in Gynostemma pentaphyllum Makino, has attracted more attention owing to its wide bioactivities. However, the effects of Gyp on esophageal cancer cells and colon cancer cells are still unknown. The present study was to investigate the possible anti-proliferative and anti-migration activity of Gyp on human colon cancer cells SW620 and esophageal cancer cells Eca-109. Cell viability was evaluated using 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Cell membrane integrity was evaluated using flow cytometry following propidium iodide staining. Apoptotic cell death was determined by nuclear 4′-6-diamidino-2-phenylindole staining. Generation of intracellular reactive oxygen species (ROS) and changes in mitochondrial membrane potential (Δ ψm) was analyzed by flow cytometry using 2′,7′-dichlorofluorescein–diacetate and rhodamine 123 staining, respectively. Wound healing assay was carried out to investigate Gyp-inhibited migration of SW620 and Eca-109 cells. The results indicated that Gyp inhibited cell proliferation and migration in SW620 and Eca-109 cells in dose- and time-dependent manner. Gyp elevated intracellular ROS level, decreased the Δ ψm, and induced apoptotic morphology such as cell shrinkage and chromatin condensation, suggesting oxidative stress and mitochondria-dependent cell apoptosis that might be involved in Gyp-induced cell viability loss in SW620 and Eca-109 cells. The findings indicate Gyp may have valuable application in clinical colon cancer and esophageal cancer treatments.

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Induction of Apoptic Features in Human Colon Cancer Cells After Exposure to Specific Vitamin Combinations

Microscopy and Microanalysis ◽

10.1017/s1431927600036953 ◽

2000 ◽

Vol 6 (S2) ◽

pp. 892-893

Author(s):

B. Brown ◽

K. R. Garvin ◽

B. G. Hughes ◽

M. D. Standing ◽

K. L. O'Neill ◽

...

Keyword(s):

Cancer Cells ◽

Human Cancer ◽

Colon Adenocarcinoma ◽

Human Colon ◽

Human Colon Cancer ◽

Colon Cells ◽

Human Cancer Cells ◽

Induction Of Apoptosis ◽

Programed Cell Death ◽

Ht 29

Proper nutrition and vitamin consumption have long been associated with lower risks for many human cancers. Data from recent studies have suggested that exposure to antioxidant vitamins and vitamin combinations can induce genetic programed cell death, apoptosis, in human cancer cells. ‘“3 In this study, we report on the induction of apoptosis in HT-29 human colon adenocarcinoma cells after exposure to low doses of 13- c/s-retinoic acid (RA) and vitamin E succinate (VES). Induction of apoptosis was seen only in cancerous colon cells and not in normal human colon cells when exposed to RA and VES in combination, but not when exposed to either vitamin alone.Cultures of HT-29 cells were seeded in 6 well plates at 10,000 cells/well and grown at 37°C, 5% C02 in RPMI 1640 medium supplemented with 10% fetal bovine serum.

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