An in vitro approach to combat multidrug resistance in Salmonella typhi and human colon cancer with Excoecaria agallocha L. extract

Abstract Background The incidence of antibacterial resistance and colon cancer is increasing in India. Antibacterial resistance and chemoresistance demand the need of developing herbal or natural chemotherapeutic agents. Our study thus, aims to determine the antibacterial and anticancer activities of the leaf extracts of the mangrove Excoecaria agallocha. Results Liquid chromatography–mass spectroscopy analysis of the ethanolic E. agallocha extracts revealed the presence of Bergenin. The plant extract fraction containing Bergenin had potent antibacterial action against a resistant strain of Salmonella typhi with an MIC value of 15.7 ± 0.04 µg/mL. Treatment of the bacteria with the plant extract made it moderately susceptible to the antibacterial drugs ampicillin, aztreonam, cefotaxime, chloramphenicol and imipenem. The plant extract caused membrane damage and disrupted the expression of a 33 kDa outer membrane protein (OmpA) in S. typhi. It was plausibly due to this mechanism of the plant extract that made the bacteria susceptible to the antibacterial drugs to a certain extent. Further, fluorescence microscopy analysis revealed the anticancer property of the extract against a human colon cancer (DLD-1) cell line by activation of Caspase-3followed by subsequent apoptosis and exhibited cytotoxicity against the cancerous cell line with an IC50 value of 17.99 ± 1.12 µg/mL. Caspase-3 activity was observed to increase in a dose-dependent manner as determined by spectrophotometric assays. Moreover, the expression of the metalloproteinase-7 (MMP-7) was significantly reduced in plant extract treated DLD-1 colon cancer cells. Conclusion The results indicate that E. agallocha is a novel source of Bergenin, and the plant extract fraction under study may be used in combination therapy along with antibacterial drugs to combat antibacterial resistance of S. typhi and also to alleviate the risks of colon cancers in human. However, further investigations may be undertaken for its therapeutic application and to explore its potential bioactivity against other bacterial strains and human cancer cell lines.

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Cell Cycle-Dependent Regulation of N-Acetylglucosaminyltransferase-III in a Human Colon Cancer Cell Line, Colo201

Archives of Biochemistry and Biophysics ◽

10.1006/abbi.1999.1577 ◽

2000 ◽

Vol 374 (1) ◽

pp. 52-58 ◽

Cited By ~ 3

Author(s):

Rujun Kang ◽

Yoshitaka Ikeda ◽

Eiji Miyoshi ◽

Wenge Wang ◽

Wei Li ◽

...

Keyword(s):

Cell Cycle ◽

Colon Cancer ◽

Cell Line ◽

Cancer Cell Line ◽

Human Colon ◽

Colon Cancer Cell Line ◽

Colon Cancer Cell ◽

Human Colon Cancer Cell ◽

Human Colon Cancer ◽

Cycle Dependent

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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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