| 郭云山,郭 浩,朱金文,胡慧敏,王晓东,郝定均.βig-h3调控间充质样运动促进骨肉瘤的转移的研究[J].,2018,(22):4207-4212 |
| βig-h3调控间充质样运动促进骨肉瘤的转移的研究 |
| βig-h3 Promotes the Metastasis of Osteosarcoma by Regulating the Mesenchymal Mode Movement |
| 投稿时间：2018-08-08 修订日期：2018-08-30 |
| DOI：10.13241/j.cnki.pmb.2018.22.002 |
| 中文关键词: βig-h3 间充质样运动 骨肉瘤 转移 |
| 英文关键词: βig-h3 Mesenchymal mode movement Osteosarcoma Metastasis |
| 基金项目:国家自然科学基金青年项目(81502330)；陕西省自然科学基金青年项目(2016JQ8040)；中国博士后科学基金特别资助项目(2017T100763)；中国博士后科学基金面上项目(2016M600805) |
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| 中文摘要: |
| 摘要 目的：明确转化生长因子β诱导基因-克隆3 (TGF-β-induced gene-human colne 3，βig-h3)在调控骨肉瘤细胞间充质样运动中的作用。方法：将骨肉瘤Saos-2细胞转染βig-h3真核表达质粒(βig-h3 Vector)和空载体(Control Vector)以增加βig-h3的表达后，应用侵袭实验、黏附实验、划痕实验和明胶酶谱实验检测βig-h3过表达对骨肉瘤细胞的侵袭能力、黏附能力、迁移能力以及基质金属蛋白酶分泌能力的影响；Western-blot实验检测Saos-2细胞的中黏着斑激酶(FAK)和桩蛋白(Paxillin)以及介导间充质样运动关键分子Rac1和WAVE2的表达水平和磷酸化水平。结果：骨肉瘤Saos-2细胞转染βig-h3真核表达质粒24小时后，细胞中βig-h3的蛋白和mRNA水平显著增加(P<0.05)。βig-h3过表达的骨肉瘤Saos-2细胞的侵袭、黏附、迁移及基质金属蛋白酶分泌能力、细胞伪足的重要成分黏着斑激酶(FAK)和桩蛋白(Paxillin)活性以及介导间充质样运动关键分子Rac1和WAVE2磷酸化水平均较空载体转染组均显著增加(P<0.05)。结论：βig-h3可能通过激活Rac1-WAVE2信号通路，促进骨肉瘤细胞侵袭、黏附、迁移、伪足形成以及基质金属蛋白酶的分泌，介导骨肉瘤细胞的间充质样运动，从而促进骨肉瘤转移。 |
| 英文摘要: |
| ABSTRACT Objective: To identify the role of transforming growth factor β-inducible gene-clone 3 (βig-h3) in the regulation of mesenchymal mode movement of osteosarcoma cells. Methods: In this study, βig-h3 eukaryotic expression plasmid (βig-h3 Vector) and empty carrier (Control Vector) were used to upregulate the expressions of βig-h3 in human osteosarcoma cells Saos-2. Cell invasion test, cell adhere test, monolayer wound healing assay and gelatin enzyme spectrum were used to test cells potential of invasion, adhesion, migration and MMPs secretion. Western-blot was used to test the phosphorylation levels of focal adhesion kinase (FAK) and pile protein (Paxillin), which were the important constituents of cell pseudo-foot and the phosphorylation levels of Rac1 and WAVE2, which were the key regulatory molecular of mesenchymal mode movement. Results: After transfection of βig-h3 eukaryotic expression plasmid (βig-h3 vector), We found that the protein and mRNA of βig-h3 were significant increased in Saos-2 cells (P<0.05). Moreover, we found that the numbers of attached cells, invaded cells, wound closure rate and MMPs secretion were significantly increased after transfection of βig-h3 eukaryotic expression plasmid (βig-h3 vector) (P<0.05). Furthermore, phosphorylated FAK, phosphorylated Paxillin, phosphorylated Rac1 and phosphorylated WAVE2 were markedly increased after transfection of βig-h3 eukaryotic expression plasmid (βig-h3 vector) (P<0.05). Conclusion: By activating the Rac1-wave2 signaling pathway, βig-h3 may regulate the mesenchymal mode movement and promotes the invasion, adhesion, migration, MMPs secretion and pseudo-foot formation in osteosarcoma cells, and thus promoting the metastasis of osteosarcoma. |
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