Siyuan Liu1,2, Lu Wang1, Wangwang Ding1, Dan Wang1, Xueting Wang1, Qianqian Luo1, Yapeng Lu3, Li Zhu4. 1. Institute of Special Environmental Medicine, Nantong University, 9 Seyuan Road, Nantong, 226019, China. 2. Center of Analysis and Testing, Nantong University, Nantong, 226019, China. 3. Institute of Special Environmental Medicine, Nantong University, 9 Seyuan Road, Nantong, 226019, China. lyplab@ntu.edu.cn. 4. Institute of Special Environmental Medicine, Nantong University, 9 Seyuan Road, Nantong, 226019, China. zhuli85051796@163.com.
Abstract
BACKGROUND: 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. METHODS: 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. RESULTS: 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. CONCLUSION: CleA suppressed the invasion of MDA-MB-231 cells, likely through the β-catenin pathway.
BACKGROUND: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-231humanbreast cancer cells was investigated, and the mechanisms involved were clarified. METHODS: 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. RESULTS: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. CONCLUSION:CleA suppressed the invasion of MDA-MB-231 cells, likely through the β-catenin pathway.
Entities:
Keywords:
Cleistanthin A; Human breast cancer cells; Invasion; β-catenin
Authors: Michal Štefánik; Dattatry Shivajirao Bhosale; Jan Haviernik; Petra Straková; Martina Fojtíková; Lucie Dufková; Ivana Huvarová; Jiří Salát; Jan Bartáček; Jan Svoboda; Miloš Sedlák; Daniel Růžek; Andrew D Miller; Luděk Eyer Journal: Viruses Date: 2022-02-09 Impact factor: 5.048