Institute of Special Environmental Medicine, Nantong University, 9 Seyuan Road, Nantong, 226019, China.
Center of Analysis and Testing, Nantong University, Nantong, 226019, China.
Pharmacol Rep. 2020 Feb;72(1):188-198. doi: 10.1007/s43440-019-00012-1. Epub 2019 Dec 19.
Cleistanthin A (CleA), a natural diphyllin glycoside, has been shown to suppress the invasion of cancer cells, but the underlying mechanisms remain unclear. Here, the inhibitory effect of CleA on the invasion of MDA-MB-231 human breast cancer cells was investigated, and the mechanisms involved were clarified.
Cell viability was studied by MTT assay. The migration and invasion of MDA-MB-231 cells were assessed by wound healing assay and transwell assay, respectively. The enzymatic activity of matrix metalloproteinases (MMPs) was detected by gelatin zymography. mRNA and protein levels were detected by qRT-PCR and Western blotting, respectively. Nuclear translocation of β-catenin was observed by immunofluorescence and detected by Western blotting.
CleA effectively inhibited the migration and invasion of MDA-MB-231 cells and suppressed the expression and activation of MMP-2/9. Moreover, the expression and nuclear translocation of β-catenin were reduced by CleA treatment, as well as transcription of the Cyclin D1 and c-myc genes. In addition, the inhibitory effect of CleA on the β-catenin pathway was attributed to the promotion of β-catenin degradation by inhibition of GSK3β phosphorylation. When the phosphorylation of GSK3β was induced by LiCl, the inhibitory effect of CleA on the β-catenin pathway and the invasion of MDA-MB-231 cells were almost reversed.
CleA suppressed the invasion of MDA-MB-231 cells, likely through the β-catenin pathway.
Cleistanthin A（CleA），一种天然的二氢菲里定糖苷，已被证明能抑制癌细胞的侵袭，但潜在的机制尚不清楚。在这里，研究了 CleA 对 MDA-MB-231 人乳腺癌细胞侵袭的抑制作用，并阐明了涉及的机制。
通过 MTT 法研究细胞活力。通过划痕愈合试验和 Transwell 试验分别评估 MDA-MB-231 细胞的迁移和侵袭。通过明胶酶谱法检测基质金属蛋白酶（MMPs）的酶活性。通过 qRT-PCR 和 Western blot 分别检测 mRNA 和蛋白水平。通过免疫荧光和 Western blot 观察β-catenin 的核转位。
CleA 有效抑制 MDA-MB-231 细胞的迁移和侵袭，并抑制 MMP-2/9 的表达和激活。此外，CleA 处理后β-catenin 的表达和核转位减少，Cyclin D1 和 c-myc 基因的转录也减少。此外，CleA 对β-catenin 通路的抑制作用归因于抑制 GSK3β 磷酸化促进β-catenin 降解。当 LiCl 诱导 GSK3β 磷酸化时，CleA 对β-catenin 通路和 MDA-MB-231 细胞侵袭的抑制作用几乎被逆转。
CleA 抑制 MDA-MB-231 细胞的侵袭，可能通过β-catenin 通路。
