收稿日期: 2023-05-24
网络出版日期: 2023-09-26
基金资助
广东省医学科研基金项目(A2021119);广州市卫生健康科技项目(20221A011098)
Preliminary study on the effects and related mechanisms of Taquimod on malignant phenotype of human glioblastoma cells
Received date: 2023-05-24
Online published: 2023-09-26
目的 探索他喹莫德（Tasquinimod，Tasq）对胶质母细胞瘤（glioblastoma multiforme，GBM）细胞增殖、迁移、上皮-间质转化（epithelial-mesenchymal transition，EMT）、干细胞特性等恶性表型的影响及相关机制，为开发Tasq作为潜在治疗GBM的药物提供研究基础。 方法 通过CCK-8法、细胞凋亡染色、Transwell实验、克隆形成实验评价Tasq对不同GBM细胞系（U87、U251）增殖、凋亡、迁移和集落形成的影响；Western blot检测凋亡、EMT等相关蛋白的表达情况。利用肿瘤干细胞成球实验检测Tasq对胶质瘤干细胞（glioma stem cell，GSC）球体形成能力的影响，免疫荧光染色和Western blot检测GSC中干细胞特性相关蛋白的表达情况。 结果 Tasq呈浓度依赖性和时间依赖性抑制U87和U251的增殖；Tasq能抑制U87和U251迁移和集落形成，诱导其发生凋亡；Tasq促进凋亡蛋白Cleaved Caspase-3表达的同时下调抗凋亡蛋白BCL-2和EMT相关蛋白（Snail、Vimentin）的表达；Tasq能显著降低GSC的成球能力及干细胞特性相关蛋白（SOX2、Nestin）的表达，与对照组相比差异均有统计学意义（P < 0.01）。 结论 Tasq能在体外抑制GBM细胞增殖、迁移、集落形成，诱导其凋亡，Tasq可能通过抑制GBM细胞的EMT和干细胞特性抑制其恶性进展。
柳晓蕊 , 袁忠民 , 郅程 , 张媚媚 , 简晓顺 , 彭怀东 . 他喹莫德对人胶质母细胞瘤细胞恶性表型的影响及相关机制[J]. 实用医学杂志, 2023 , 39(16) : 2029 -2036 . DOI: 10.3969/j.issn.1006-5725.2023.16.004
Objective To investigate the effects and related mechanisms of Tasquinimod （Tasq） on malignant phenotypes such as proliferation， migration， epithelial-mesenchymal transition （EMT）， stem cell characteristics in glioblastoma multiforme（GBM）cells， so as to provide reference for preliminary experiment of Tasq as a potential drug treating GBM. Methods The effects of Tasq on the proliferation， apoptosis， migration and colony formation of GBM cell lines （U87 and U251） were evaluated by CCK?8， apoptosis staining， transwell assay and colony formation assay. The expression of apoptosis?related proteins and EMT?related proteins were detected by western blot. The effect of Tasq on spheres formation of glioma stem cell （GSC） was detected by sphere formation assay， and the expression of stem cell?related proteins in GSC were detected by immunofluorescence staining and Western blot. Results Tasq inhibited the proliferation of U87 and U251 in concentration?dependent and time?dependent manner； Tasq also inhibited the migration and colony formation of U87 and U251， and promoted its apoptosis；Tasq promoted the expression of apoptotic protein Cleaved Caspase?3 and decreased the expression of anti?apoptotic protein BCL?2， EMT?related proteins （Snail，Vimentin）. Tasq also reduced the spheroid formation ability of GSC and the expression of stem cell?related proteins （SOX2，Nestin）， the differences were statistically significant while compared with that in the control group（P < 0.01）. Conclusion Tasq inhibits proliferation， migration， colony formation of GBM cells and promoted its apoptosis in vitro. Tasq may inhibit the malignant progression of GBM cells by inhibiting EMT and stem cell characteristics.
Key words： Tasquinimod; glioblastoma cell; EMT; stem cell characteristics
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