Background: Nostoc commune Vauch. is a nitrogen-fixing blue-green algae that expresses a large number of active molecules with medicinal properties. Our previous study found that a water stress protein (WSP1) from N. commune and its recombinant counterpart (Re-WSP1) exhibited significant anti-colon cancer activity both in vitro and in vivo. This study is to investigate the effects of Re-WSP1 on proliferation of colon cancer cells and to elucidate the relevant mechanisms.
Methods: Real-time quantitative PCR was used to detect the expression of miR-539 in colon cancer HT-29 and DLD1 cells. Colon cancer cells were transfected with miR-539 mimics and negative controls, and cell proliferation were detected by CCK8 and clonogenic assays. The target gene of miR-539 was predicted, and the dual luciferase reporter gene experiment was used to verify the target gene. After colon cancer cells were transfected with miR-539 mimics or inhibitors, the expression of target gene β-catenin was detected by Western blot. miR-539 inhibitor confirmed cell proliferation.
Results: Re-WSP1 inhibited colon cancer cell growth in a dose-dependent manner. Re-WSP1 inhibited the expression of β-catenin, which was partly reversed by LiCl treatment. Quantitative PCR analysis showed that the expression of miR-539 was significantly upregulated after Re-WSP1 treatment. Moreover, miR-539 negatively regulated the expression of β-catenin by directly binding to the 3'UTR of β-catenin mRNA. The cell growth inhibition and the decrease in β-catenin expression induced by Re-WSP1 were significantly reversed by miR-539 inhibitor.
Conclusion: Re-WSP1 suppresses colon cancer cell growth via the miR-539/β-catenin axis.
Keywords: Colon cancer; Nostoc commune Vauch.; Re-WSP1; microRNA-539; β-catenin.
© 2021. The Author(s), under exclusive licence to Springer Nature B.V.