Effect of indomethacin on E-cadherin and beta-catenin expression in HT-29 colon cancer cells.
ABSTRACT Nonsteroidal anti-inflammatory drugs (NSAIDs) lower the incidence of and mortality from colon cancer. Although there is much evidence from epidemiological and laboratory studies that NSAIDs have antitumor activity and reduce the incidence of colon cancer, the mechanism of action remains unknown. In this paper, we present the effect of indomethacin on growth inhibition, induction of apoptosis, and alterations in the expression of several genes involved in Wnt signaling in HT-29 colon cancer cells. We have shown that indomethacin reduces the proliferation rate of HT-29 colon cancer cells and induces apoptosis. Concentrations of indomethacin from 10(-4) to 10(-3) M strongly inhibited the growth of HT-29 cells. The inhibition of growth, as well as induction of apoptosis was dose and time dependent. The treatment of cells with 4 x 10(-4) M indomethacin caused strong inhibition of cell growth (about 70%), enhanced expression of APC, decreased expression of beta-catenin and induced expression of E-cadherin proteins. Expression of beta-catenin was not markedly reduced instead, beta-catenin was translocated from the nucleus and cytoplasm to the plasma membrane. These results were confirmed by real-time RT-PCR analysis on mRNA level. At a concentration of 4 x 10(-4) M indomethacin there was increased expression of APC gene (10.9-fold induction; DeltaDeltaCt = 3.43) and E-cadherin gene (3.5-fold induction; DeltaDeltaCt = 1.79). These results suggest the antiproliferative effect of indomethacin may contribute to enhanced cell adhesion through increased expression of E-cadherin and translocation of beta-catenin from the nucleus to the cell membrane.
SourceAvailable from: Velichka AndonovaWorld Journal of Pharmaceutical Research 01/2015; 4(1):85-101. · 5.99 Impact Factor
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ABSTRACT: Background: In the previous study it was shown that films prepared from inulin (In) in combination with Eudragit RS (ERS) and RL (ERL) were susceptible to inulinase. Purpose: The aim of this work was to assess the suitability of these combinations for colonic delivery of indomethacin. Methods: Indomethacin was loaded onto non-pareil seeds using fluidized bed apparatus to produce pellets with 20% w/w drug load. Drug loaded pellets were coated with In-ERS in the ratios of 20:80 and 30:70, or In-ERL in the ratio of 20:80 to different coating loads. The release of drug was examined in simulated gastric (for 2 hrs) and small intestine and in the presence of inulinase in simulated colonic medium (for 12 or 24 hrs). Results: The results of this study revealed that incorporation of inulin as a bacterially degradable polysaccharide into ERS or ERL could modulate drug release. Coating level up to 15% significantly affected drug release from In-ERL or In-ERS coated pellets. However further increase in coating load to 20% had no significant effect on drug release from In-ERL coated pellets (f1=9.39). Drug release from In-ERL coated pellets was faster and showed some pH dependency. Conclusions: Formulation coated with In-ERS (20:80) and coating level of 20% was considered more appropriate for colon delivery of indomethacin, as drug release was pH independent and formulation was resistant to drug release in the upper GI media for up to 7 hrs. This formulation was also susceptible to inulinase and released about 40% of indomethacin in the simulated colonic media.
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ABSTRACT: Indometacin, an inhibitor of cyclooxygenase-2 (COX-2), has been shown to exert anticancer effects in a variety of cancers. However, the effect and mechanism of indometacin on high glucose (HG)-induced proliferation and invasion of pancreatic cancer (PC) cells remain unclear. Multiple lines of evidence suggest that a large portion of pancreatic cancer (PC) patients suffer from either diabetes or HG which contributing PC progression. In this study, we report that indometacin down-regulated HG-induced proliferation and invasion via up-regulating E-cadherin but not COX-2 in PC cells. Additionally, the E-cadherin transcriptional repressors, Snail and Slug, were also involved in the process. In addition, the proliferation and invasion of PC cells, incubated in HG medium and treated with indometacin were significantly increased when E-cadherin was knocked down (Si-E-cad). Moreover, the protein levels of MMP-2, MMP-9 and VEGF were increased in PC cells transfected with Si-E-cad. Finally, the activation of the PI3K/AKT/GSK-3β signaling pathway was demonstrated to be involved in indometacin reversing HG-induced cell proliferation and invasion in PC cells. In conclusion, these results suggest that indometacin plays a key role in down-regulating HG-induced proliferation and invasion in PC cells. Our findings indicate that indometacin could be used as a novel therapeutic strategy to treat PC patients who simultaneously suffer from diabetes or HG.Current Medicinal Chemistry 08/2013; DOI:10.2174/09298673113209990249 · 3.72 Impact Factor