Background: Targeting DNA mismatch repair-deficient/KRAS-mutant Colorectal Cancer Stem Cells (CRCSCs) with chemical compounds remains challenging. Modulating stemness factors Bmi-1, Sox-2, Oct-4 and Nanog in CRCSCs which are direct downstream targets of carcinogenesis pathways may lead to the reactivation of caspase-3 and apoptosis in these cells. Omega-3 DHA modulates different signaling pathways involved in carcinogenesis. However, little is known, whether in vitro concentrations of DHA equal to human plasma levels are able to modulate pluripotency genes expression, caspase-3 reactivation and apoptosis in DNA mismatch repair-deficient/KRAS-mutant CRC stem-like cells.
Methods: DNA mismatch repair-deficient/KRAS-mutant CRC stem-like cells (LS174T cells) were treated with DHA, after which, cell number and proliferation-rate, Bmi-1, Sox-2, Nanog and Oct-4 expression, caspase-3 activation and apoptosis were evaluated with different cellular and molecular techniques.
Results: DHA changed the morphology of cells to apoptotic forms and disrupted cell connections. After 48h treatment with 50- to 200μM DHA, cell numbers and proliferation-rates were measured to be 86%-35% and 93.6%-45.7% respectively. Treatment with 200 μM DHA dramatically decreased the expression of Bmi-1, Sox- 2, Oct-4 and Nanog by 69%, 70%, 97.5% and 53% respectively. Concurrently, DHA induced caspase-3 activation by 1.8-4.7-fold increases compared to untreated cells. An increase in the number of apoptotic cells ranging from 9.3%-38.4% was also observed with increasing DHA concentrations.
Conclusions: DHA decreases the high expression level of pluripotency network genes suggesting Bmi-1, Sox-2, Oct-4 and Nanog as promising molecular targets of DHA. DHA reactivates caspase-3 and apoptosis in DNA mismatch repair-deficient/KRAS-mutant CRC stem-like cells, representing the high potential of this safe compound for therapeutic application in CRC.
[http://dx.doi.org/10.1056/NEJM199805213382101] [PMID: 9593786]
[http://dx.doi.org/10.1038/nature11156] [PMID: 22722830]
[http://dx.doi.org/10.1007/s00432-009-0745-7] [PMID: 20024662]
[http://dx.doi.org/10.1371/journal.pone.0041335] [PMID: 22912670]
[http://dx.doi.org/10.1007/s00595-018-1644-9] [PMID: 29488015]
[http://dx.doi.org/10.1016/S0092-8674(03)00393-3] [PMID: 12787504]
[http://dx.doi.org/10.1016/S0304-3835(02)00432-9] [PMID: 12359365]
[http://dx.doi.org/10.1080/07315724.2011.10719969] [PMID: 21917707]
[http://dx.doi.org/10.3109/07357907.2012.743553] [PMID: 23193970]
[http://dx.doi.org/10.1093/embo-reports/kvf044] [PMID: 11850399]
[http://dx.doi.org/10.1038/nm.3418] [PMID: 24292392]
[http://dx.doi.org/10.1073/pnas.96.5.2316] [PMID: 10051639]
[http://dx.doi.org/10.1073/pnas.0703478104] [PMID: 17548814]
[http://dx.doi.org/10.1080/01635581.2017.1247884] [PMID: 27880058]
[http://dx.doi.org/10.1016/j.jnutbio.2014.06.013] [PMID: 25277647]
[http://dx.doi.org/10.1089/cbr.2018.2445] [PMID: 30395490]
[http://dx.doi.org/10.1016/j.jnutbio.2012.03.023] [PMID: 22854319]
[http://dx.doi.org/10.1007/s13402-015-0254-4] [PMID: 26671842]
[http://dx.doi.org/10.1172/JCI34401] [PMID: 18497886]
[http://dx.doi.org/10.1038/sj.bjc.6605762] [PMID: 20606680]