高良姜素通过影响Hippo/YAP通路抑制宫颈癌Hela细胞迁移和侵袭

严怡然; 沈成万; 尚香玉; 冯婵; 李金秋; 阿仙姑·哈斯木

高良姜素通过影响Hippo/YAP通路抑制宫颈癌Hela细胞迁移和侵袭

1.
新疆医科大学基础医学院/新疆地方病分子生物学重点实验室，新疆乌鲁木齐　836001
2.
宁德师范学院医学院，福建宁德　352100

基金项目: 福建省自然科学基金（2021J011174）；宁德师范学院科技项目（2021Q105）

详细信息

作者简介:
严怡然（1997～），女，新疆乌鲁木齐人，在读硕士研究生，主要从事宫颈癌的发病机制研究工作

通讯作者:
阿仙姑·哈斯木，E-mail：axiangu75@126.com

中图分类号: R737.33
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出版历程
- 收稿日期: 2024-09-04

Galangin Inhibits the Migration and Invasion of Cervical Cancer Hela Cells Through Hippo/YAP Pathway

1.
School of Basic Medicine / Xinjian Key Laboratory of Molecular Biology of Endemic Diseases，Urumqi Xinjiang 836001
2.
School of Medicine，Ningde Normal University，Ningde 352100，China

摘要

摘要: 目的探讨高良姜素对宫颈癌Hela细胞迁移、侵袭能力的影响及其潜在机制。方法用不同浓度的高良姜素（0、5、10、20、40、60、80、100 µmol/L）干预宫颈癌细胞48 h，采用 CCK-8 实验检测高良姜素对细胞活力的影响及筛选高良姜素的半数致死量；将Hela细胞分为对照组（0 μmol/L）、高良姜素组（40 μmol/L处理）。划痕愈合实验和Transwell小室法分别检测各组细胞的迁移和侵袭能力，Western blot法检测各组细胞中E-cadherin、 N-cadherin蛋白的表达；DIA定量蛋白质组学技术检测并筛选两组之间的差异表达蛋白质；利用KEGG Pathway、基因集富集分析（GSEA）方法对差异基因进行生物学功能富集分析；Western blot法验证富集的Hippo/YAP信号通路相关蛋白YAP和p-YAP表达水平。结果与对照组（0 μmol/L）比较，高良姜素以浓度依赖性方式抑制宫颈癌Hela细胞活性（P < 0.05；P < 0.001）；与对照组（0 μmol/L）比较，高良姜素（40 μmol/L）干预后宫颈癌Hela细胞的划痕愈合能力和侵袭能力明显降低（P < 0.001）；与对照组（0 μmol/L）比较，高良姜素组（40 μmol/L）E-cadherin蛋白表达升高（P < 0.05）、 N-cadherin 蛋白表达降低（P < 0.001）；KEGG和GSEA富集结果发现，高良姜素抑制宫颈癌恶性进展与Hippo/YAP信号通路显著相关，Western blot验证发现Hippo信号通路中标志性蛋白p-YAP蛋白表达水平升高（P < 0.01），YAP蛋白表达降低（P < 0.05）。结论高良姜素作用于Hela 细胞后，抑制细胞的增殖、迁移和侵袭能力，并表现出剂量依赖；其机制可能与Hippo/YAP信号通路激活相关。
- 高良姜素 /
- 宫颈癌 /
- 迁移 /
- 侵袭 /
- Hippo/YAP信号通路
Abstract: Objective To investigate the effects of galangin on the migration and invasion abilities of cervical cancer Hela cells and its potential mechanisms. Methods Hela cells were treated with different concentrations of galangin （0, 5, 10, 20, 40, 60, 80, 100 µmol/L） for 48 hours, and CCK-8 assay was used to assess the impact of galangin on cell viability and to determine the half-maximal lethal concentration （LC50） of galangin. Hela cells were divided into a control group （0 μmol/L） and a galangin group （40 μmol/L treatment）. Scratch wound healing assays and Transwell chamber assays were conducted to evaluate the migration and invasion abilities of the cells in each group. Western Blot was used to detect the protein expression of E-cadherin and N-cadherin. DIA quantitative proteomics technology was used to detect and screen the differentially expressed proteins between the two groups. Biological function enrichment analysis of the differential genes was performed using the Kyoto Encyclopedia of Genes and Genomes （KEGG） Pathway and Gene Set Enrichment Analysis （GSEA） methods. Western Blot was used to verify the expression levels of Hippo/YAP signaling pathway-related proteins YAP and p-YAP. Results Compared to the control group （0 μmol/L）, galangin （40 μmol/L） significantly inhibited the viability of Hela cells in a concentration-dependent manner （P < 0.05; P < 0.001）. Compared with the control group （0 μmol/L）, the scratch healing ability and invasion ability of cervical cancer Hela cells treated with galangin（40 μmol/L） were significantly reduced （P < 0.001）. The expression of E-cadherin protein was increased （P < 0.05） and the expression of N-cadherin protein was decreased （P < 0.001） in the galangin group （40 μmol/L） compared to the control group （0 μmol/L）. KEGG and GSEA enrichment results indicated that the inhibition of malignant progression in cervical cancer by galangin was significantly associated with the Hippo/YAP signaling pathway. Western Blot confirmed that the expression level of the hallmark protein p-YAP in the Hippo signaling pathway was increased （P < 0.01）, while the expression level of YAP protein was decreased （P < 0.05）. Conclusion Galangin inhibits the proliferation, migration and invasion abilities of Hela cells in a dose-dependent manner. The underlying mechanism might be associated with the activation of the Hippo/YAP signaling pathway.
- Galangin /
- Cervical cancer /
- Migration /
- Invasion /
- Hippo/YAP signaling pathway

HTML全文

图 1 高良姜素对宫颈癌Hela细胞增殖的影响

CCK-8法测定不同浓度高良姜素处理Hela细胞对细胞增殖的影响。与0 µmol/L比较，^*P < 0.05；^***P < 0.001。

Figure 1. Effect of galangin on proliferation of cervical cancer Hela cells

下载: 全尺寸图片幻灯片

图 2 高良姜素对宫颈癌Hela细胞迁移和侵袭的影响

A：细胞划痕实验结果（标尺为100 μm）；B：Transwell实验结果（标尺为100 μm）；与0 mol/L组比较，^***P < 0.001。

Figure 2. Effect of galangin on migration and invasion of cervical cancer Hela cells

下载: 全尺寸图片幻灯片

图 3 高良姜素对EMT相关蛋白表达的影响

A：Western blot实验检测Hela细胞EMT相关N-cadherin、E-cadherin蛋白表达情况；B：N-cadherin蛋白相对表达量量化图；C：E-cadherin蛋白相对表达量量化图。与0 mol/L组比较，^*P < 0.05；^***P < 0.001。

Figure 3. Effect of galangin on expression of EMT-related protein

下载: 全尺寸图片幻灯片

图 4 KEGG与GSEA-GO ES富集图

A：KEGG通路的气泡图；B：GSEA-GO ES富集图中展示。

Figure 4. Enrichment diagram of KEGG and GSEA-GO ES

下载: 全尺寸图片幻灯片

图 5 高良姜素对Hippo/YAP相关蛋白表达的影响

A：Western blot实验检测Hela细胞Hippo相关Yap、Phospho-Yap蛋白表达情况；B：Yap蛋白相对表达量量化图；C：Phospho-Yap蛋白相对表达量量化图。与0 mol/L组比较，^*P < 0.05；^***P < 0.001。

Figure 5. Effect of galangin on Hippo/YAP related protein expression

下载: 全尺寸图片幻灯片

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