Nephrology Department, Shanxi Provincial People's Hospital, 29 Twin Pagoda Temple Street, Taiyuan, China.
School of Life Science, Shanxi University, 92 Wucheng Road, Taiyuan, Shanxi, China.
Mol Biol Rep. 2021 Nov;48(11):7059-7065. doi: 10.1007/s11033-021-06549-w. Epub 2021 Oct 1.
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.
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.
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.
Re-WSP1 suppresses colon cancer cell growth via the miR-539/β-catenin axis.
普通念珠藻 Vauch. 是一种固氮蓝藻，表达大量具有药用特性的活性分子。我们之前的研究发现，普通念珠藻的一种水胁迫蛋白（WSP1）及其重组对应物（Re-WSP1）在体外和体内均表现出显著的抗结肠癌活性。本研究旨在探讨 Re-WSP1 对结肠癌细胞增殖的影响，并阐明相关机制。
实时定量 PCR 检测结肠癌 HT-29 和 DLD1 细胞中 miR-539 的表达。转染 miR-539 模拟物和阴性对照物，通过 CCK8 和集落形成实验检测结肠癌细胞的增殖。预测 miR-539 的靶基因，并使用双荧光素酶报告基因实验验证靶基因。转染 miR-539 模拟物或抑制剂后，通过 Western blot 检测靶基因 β-catenin 的表达。用 miR-539 抑制剂验证细胞增殖。
Re-WSP1 呈剂量依赖性抑制结肠癌细胞生长。Re-WSP1 抑制 β-catenin 的表达，LiCl 处理部分逆转了这一作用。定量 PCR 分析显示，Re-WSP1 处理后 miR-539 的表达明显上调。此外，miR-539 通过直接结合 β-catenin mRNA 的 3'UTR 负调控 β-catenin 的表达。Re-WSP1 诱导的细胞生长抑制和 β-catenin 表达降低，被 miR-539 抑制剂显著逆转。
Re-WSP1 通过 miR-539/β-catenin 轴抑制结肠癌细胞生长。
