| 陈恩就,周方联,陈胜,郑哲D.二氢杨梅素通过Wnt/β-catenin通路抑制肺癌干细胞的研究[J].浙江中西医结合杂志,2020,30(4): |
| 二氢杨梅素通过Wnt/β-catenin通路抑制肺癌干细胞的研究 |
| The anti-tumor effects of dihydromyricetin on lung cancer stem cells by Wnt/β-catenin pathway |
| 投稿时间：2019-09-28 修订日期：2020-03-26 |
| DOI： |
| 中文关键词: 二氢杨梅素 肺癌干细胞 β-catenin |
| 英文关键词:dihydromyricetin lung cancer stem cells β-catenin |
| 基金项目:] 浙江省医药卫生科技计划项目( 2014-KYB-043) |
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| 中文摘要: |
| [摘要] 目的：分析二氢杨梅素（dihydromyricetin, DMY）对体外肺癌干细胞的作用，并明确其具体作用途径。方法：体外肺癌细胞株PC9和H1299以及人正常支气管上皮细胞株16HBE经DMY作用后，CCK-8法检测细胞活力；流式细胞术测定细胞凋亡；无血清培养基培养亲代PC9和H1299细胞建立癌干细胞（cancer stem cells, CSCs）；单细胞成球实验测定DMY对PC9/CSCs和H1299/CSCs悬浮细胞球形态和自我更新能力的影响；利用蛋白质印迹法测定Nanog、OCT4、SOX2和Wnt/β-catenin通路相关蛋白的含量。结果：（5~100 μmol/L）的DMY对16HBE细胞活力无明显抑制效应，该浓度范围的DMY呈浓度依赖性地抑制体外PC9和H1299细胞活力，（25 μmol/L）的DMY可显著诱导体外肺癌细胞凋亡；DMY可显著抑制CSCs体外单细胞成球体积和数量，并抑制Wnt/β-catenin通路活性，同时降低CSCs中Nanog、OCT4、SOX2的表达。结论：DMY在体外可抑制肺CSCs生长，其抗癌作用至少部分通过抑制Wnt/β-catenin通路活性来实现。 |
| 英文摘要: |
| [ABSTRACT] AIM: To explore the anti-tumor effects and the underlying molecular mechanisms of dihydromyricetin (DMY) on lung cancer stem cells (CSCs) in vitro. METHODS: Two human lung cancer cell lines (PC9 and H1299) as well as the normal human bronchial epithelial cell line 16HBE were exposed to certain concentration of DMY, CCK-8 was employed to detect the cell viability in vitro. Annexin V/PI apoptosis assay was utilized to explore the apoptosis of lung cancer cells. PC9/CSCs and H1299/CSCs were obtained by serum-free medium cultivation. Western blot was used to evaluate the expression of Nanog, OCT4, SOX2 and Wnt/β-catenin pathway relative factors. Single cell sphere formation assay was employed to detect the self-renewal?ability of CSCs. RESULTS: DMY exerted no cytotoxic effects on the growth of 16HBE cells from 5 to 100 μmol/L. However, DMY (5 to 100 μmol/L) treatment dose-dependently inhibited the viability of PC9 and H1299 cells. DMY triggered apoptosis of both PC9 and H1299 cells. The number and size of the tumor sphere were significantly decreased by the treatment of DMY. The activity of Wnt/β-catenin signaling pathway in both CSCs were inhibited by DMY treatment, which was consistent with the reduction of Nanog, OCT4 and SOX2 expression. CONCLUSION: DMY inhibits the stem cell properties of CSCs in vitro. And the anti-cancer function of DMY, at least partly, is realized by inactivating Wnt/β-catenin signaling. |
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