Development and characterization of an in vitro model of colorectal adenocarcinoma with MDR phenotype

Cinci, Lorenzo; Luceri, Cristina; Bigagli, Elisabetta; Carboni, Ilaria; Paccosi, Sara; Parenti, Astrid; Guasti, Daniele; Coronnello, Marcella

doi:10.1002/cam4.694

Cited by 13 publications

(10 citation statements)

References 30 publications

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“…The expression of P-gp gene in our Caco-2 cells was verified by RT-PCR. As shown in Figure 5, the cells used in the present experiment showed expression levels comparable to that of positive control (HCT-8 cancer cell line) [58].…”

Section: Resultsmentioning

confidence: 92%

Formulation of Nanomicelles to Improve the Solubility and the Oral Absorption of Silymarin

et al. 2019

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Two novel nanomicellar formulations were developed to improve the poor aqueous solubility and the oral absorption of silymarin. Polymeric nanomicelles made of Soluplus and mixed nanomicelles combining Soluplus with d-α-tocopherol polyethylene glycol 1000 succinate (vitamin E TPGS) were prepared using the thin film method. Physicochemical parameters were investigated, in particular the average diameter, the homogeneity (expressed as polydispersity index), the zeta potential, the morphology, the encapsulation efficiency, the drug loading, the critical micellar concentration and the cloud point. The sizes of ~60 nm, the narrow size distribution (polydispersity index ≤0.1) and the encapsulation efficiency >92% indicated the high affinity between silymarin and the core of the nanomicelles. Solubility studies demonstrated that the solubility of silymarin increased by ~6-fold when loaded into nanomicelles. Furthermore, the physical and chemical parameters of SLM-loaded formulations stored at room temperature and in refrigerated conditions (4 °C) were monitored over three months. In vitro stability and release studies in media miming the physiological conditions were also performed. In addition, both formulations did not alter the antioxidant properties of silymarin as evidenced by the 1,1-Diphenyl-2-picrylhydrazyl radical (DPPH) assay. The potential of the nanomicelles to increase the intestinal absorption of silymarin was firstly investigated by the parallel artificial membrane permeability assay. Subsequently, transport studies employing Caco-2 cell line demonstrated that mixed nanomicelles statistically enhanced the permeability of silymarin compared to polymeric nanomicelles and unformulated extract. Finally, the uptake studies indicated that both nanomicellar formulations entered into Caco-2 cells via energy-dependent mechanisms.

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Section: Resultsmentioning

confidence: 92%

Formulation of Nanomicelles to Improve the Solubility and the Oral Absorption of Silymarin

et al. 2019

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“…Cell migration was assessed with the modified Boyden chamber (48-multiwell plates; Neuroprobe, Gaithersburg, MD, USA) as previously reported [ 58 ]. Briefly, 1% fetal calf serum (FCS)-containing medium alone (control) or supplemented with 25 μM dex was added to the lower wells, while 20 × 10 4 cells were seeded into the upper wells without or with RU486, 680C91, or actinomycin D and incubated at 37 °C for 24 h. Methanol-fixed cells were stained with Diff-Quik (Dade Behring, Dudingen, Switzerland), and cell migration was measured by microscopic evaluation of the number of cells moved across the filter in ten randomly selected fields at magnification 400×.…”

Section: Methodsmentioning

confidence: 99%

Dexamethasone Induces the Expression and Function of Tryptophan-2-3-Dioxygenase in SK-MEL-28 Melanoma Cells

et al. 2021

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Tryptophan-2,3-dioxygenase (TDO) is one of the key tryptophan-catabolizing enzymes with immunoregulatory properties in cancer. Contrary to expectation, clinical trials showed that inhibitors of the ubiquitously expressed enzyme, indoleamine-2,3-dioxygenase-1 (IDO1), do not provide benefits in melanoma patients. This prompted the hypothesis that TDO may be a more attractive target. Because the promoter of TDO harbors glucocorticoid response elements (GREs), we aimed to assess whether dexamethasone (dex), a commonly used glucocorticoid, modulates TDO expression by means of RT-PCR and immunofluorescence and function by assessing cell proliferation and migration as well as metalloproteinase activity. Our results show that, in SK-Mel-28 melanoma cells, dex up-regulated TDO and its downstream effector aryl hydrocarbon receptor (AHR) but not IDO1. Furthermore, dex stimulated cellular proliferation and migration and potentiated MMP2 activity. These effects were inhibited by the selective TDO inhibitor 680C91 and enhanced by IDO1 inhibitors. Taken together, our results demonstrate that the metastatic melanoma cell line SK-Mel-28 possesses a functional TDO which can also modulate cancer cell phenotype directly rather than through immune suppression. Thus, TDO appears to be a promising, tractable target in the management or the treatment of melanoma progression.

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“…Adherent HCT-8 cells, metastatic cells and adherent new colonies were treated for 72 h with 5-fluorouracil (5FU) 10 −6 M 14 and with a FOLFOX-like treatment (5FU 10 −6 M + Oxaliplatin 2 × 10 −7 M) (Sigma Aldrich, Milan, Italy). Cells viability was assessed by MTS assay (Promega Corporation, WI).…”

Section: Methodsmentioning

confidence: 99%

Exosomes secreted from human colon cancer cells influence the adhesion of neighboring metastatic cells: Role of microRNA-210

Bigagli

Luceri

Guasti

et al. 2016

Cancer Biology & Therapy

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Cancer-secreted exosomes influence tumor microenvironment and support cancer growth and metastasis. MiR-210 is frequently up-regulated in colorectal cancer tissues and correlates with metastatic disease. We investigated whether exosomes are actively released by HCT-8 colon cancer cells, the role of exosomal miR-210 in the cross-talk between primary cancer cells and neighboring metastatic cells and its contribution in regulating epithelial-mesenchymal transition (EMT) and mesenchymal-epithelial transition (MET).After 7 d of culture, a subpopulation of viable HCT-8 cells detached the monolayer and started to grow in suspension, suggesting anoikis resistance and a metastatic potential. The expression of key proteins of EMT revealed that these cells were E-cadherin negative and vimentin positive further confirming their metastatic phenotype and the acquisition of anoikis resistance. Metastatic cells, in the presence of adherently growing HCT-8, continued to grow in suspension whereas only if seeded in cell-free wells, were able to adhere again and to form E-cadherin positive and vimentin negative new colonies, suggesting the occurrence of MET.The chemosensitivity to 5 fluorouracil and to FOLFOX-like treatment of metastatic cells was significantly diminished compared to adherent HCT-8 cells. Of note, adherent new colonies undergoing MET, were insensitive to both chemotherapeutic strategies. Electron microscopy analysis demonstrated that adherently growing HCT-8, actually secreted exosomes and that exosomes in turn were taken up by metastatic cells. When exosomes secreted by adherently growing HCT-8 were administered to metastatic cells, MET was significantly inhibited. miR-210 was significantly upregulated in exosomes compared to its intracellular levels in adherently growing HCT-8 cells and correlated to anoikis resistance and EMT markers.Exosomes containing miR-210 might be considered as EMT promoting signals that preserve the local cancer-growth permissive milieu and also guide metastatic cells to free, new sites of dissemination.

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Development and characterization of an in vitro model of colorectal adenocarcinoma with MDR phenotype

Cited by 13 publications

References 30 publications

Formulation of Nanomicelles to Improve the Solubility and the Oral Absorption of Silymarin

Formulation of Nanomicelles to Improve the Solubility and the Oral Absorption of Silymarin

Dexamethasone Induces the Expression and Function of Tryptophan-2-3-Dioxygenase in SK-MEL-28 Melanoma Cells

Exosomes secreted from human colon cancer cells influence the adhesion of neighboring metastatic cells: Role of microRNA-210

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