PO-041 Tnf pathway in metastatic colorectal cancer according to ras status and implication of potassium channels

S IBRAHIM, A Girault, L Babin, M Guéguinou, M Potier-Cartereau, C Vandier, G Paintaud, T Lecomte, W Raoul
2018 Poster Presentation: Cancer Cell Biology   unpublished
Introduction Novel marine compounds supposed an unexplored source of new drugs and expand the possibilities of finding active molecules in all the different signatures of cancer, such as proliferation, invasion and metastases. Here we report the capacity of four marine invertebrate extracts that inhibit proliferation, invasion and migration in colon cancer cell model. Material and methods HCT116 cell line was cultured and exposed to four marine invertebrate extracts (CR from a red coral, PS
more » ... an holothurian and NA and NB from nudibranchs) at different concentrations for 24 hour and IC50 was calculated by MTT assay to estimate cytotoxic activity. The rate of cell proliferation and migration was monitored in realtime with the xCELLigence system with E and CIM-16 plates, respectively (Roche Diagnostics GmbH, Germany), which were set in a humidified incubator and maintained in 5% CO2 at 37°C. The impedance value of each well was automatically monitored by the xCELLigence system for 72 hour and expressed as a cell index (CI) value. The anti-invasive rate was determined by Corning BioCoat Matrigel Invasion Chambers with 8.0 mm PET Membrane 24-well Plates (Corning, NY, USA), and crystal violet assay was used to calculate CI. Results and discussions The four marine invertebrate extracts exhibited pronounced cytotoxic and antiproliferative effect in HCT116 model at different doses. The migration assays revealed that CR and especially NB extracts reduced the migration ability of HCT116 cells. Invasion assay let us to isolate superinvasive populations derived from HCT116 which lead to cell models bearing different invasive capabilities (parental (P); invasive subpopulation cells (invaded through membrane four times (I4); and superinvasive line 9 (I9)). Invasive subpopulations I4 and I9 presented a higher rate of invasion than parental population. NB extract showed the best antimigratory capacity compared to other extracts and therefore was selected for further invasive assays. NB showed to reduce invasive cell index in all superinvasive populations. Conclusion The results support the antiproliferative, antimigration and anti-invasive activity of CR, PS, NA and NB marine invertebrate extracts in the highly invasive colon cancer model HCT116. Due to its strong activity, further studies will be focused on the potential of NB extract in the inhibition of metastases-related processes. Introduction Marine invertebrate organisms have received great attention from the scientific community due to its richness in new bioactive compounds with potential anti-proliferative and anticancer activities. In the present study, the cytotoxic efficacy of four marine invertebrate-extracts against human adenocarcinoma colon cancer cells was investigated. Material and methods HT29, SW480 and HGUE-C-1 human colon adenocarcinoma cell lines were cultured and exposed to 4 marine invertebrate extracts at different concentrations for 24 hour. The anti-proliferative properties were examined by MTT assay. Analysis of apoptosis and cell cycle stages were performed with the Muse Cell Analyzer (Millipore, Hayward, CA, USA). Mitochondrial functionality was determined by the ratio of MitoTracker Red CMXRos and MitoTracker Green FM fluorescent probes (Molecular Probes). Measurement of lactate dehydrogenase (LDH) was performance by a colorimetric assay (Roche Diagnostic, Germany). Results and discussions The four marine invertebrate extracts exhibited pronounced cytotoxic effects in the three cancer cell lines used. In this study, CR, PS, NA and NB extracts were able to modify the cell cycle of HGUE-C-1, HT-29 and SW-480 cells, by increasing cell population in the subG1 and G2/ M phase in a dose-dependent manner. Results confirmed that NA, NB and CR extracts induced an increase in the percentage of cell population in early apoptosis by Annexin V staining. PS extract on the contrary, raised the late apoptotic population at different concentrations. Mitochondrial membrane depolarization rate, as indicator of apoptosis, was induced significantly by CR, NA and NB extracts in the three cell models used. In contrast PS reduced this parameter in a dose-dependent manner. Finally, the release of the enzyme lactate dehydrogenase (LDH) confirmed a cell death by a necrotic pathway for PS extract. Conclusion Our results strongly suggest that NB, NA and CR extracts exhibit antiproliferative and pro-apoptotic properties in colon cancer cell lines concomitantly to a decreased mitochondrial membrane potential. In contrast, PS extract shows an antiproliferative effect mediated by necrosis in the same cell models. This preliminary study suggests that these extracts present pharmacological potential. Further investigations to determine the responsible compounds and the underlying mechanism are currently ongoing.
doi:10.1136/esmoopen-2018-eacr25.89 fatcat:4xqw3mnewna3vlfdzegth6zpue