苦马豆素通过降低 miR-92a 的表达来抑制神经胶质瘤细胞的增殖、迁移和侵袭。
Swainsonine represses glioma cell proliferation, migration and invasion by reduction of miR-92a expression.
机构信息
Department of Neurosurgery, China-Japan Union Hospital of Jilin University, No.126, Xiantai Street, Changchun, 130033, Jilin Province, China.
Department of Vascular Surgery, China-Japan Union Hospital of Jilin University, Changchun, 130033, Jilin Province, China.
出版信息
BMC Cancer. 2019 Mar 19;19(1):247. doi: 10.1186/s12885-019-5425-7.
BACKGROUND
Swainsonine is a natural indolizidine alkaloid, its anti-tumor activity has been widely reported in varied cancers. This study aimed to investigate whether Swainsonine exerted anti-tumor impact on glioma cells, likewise uncovered the relative molecular mechanisms.
METHODS
After administration with diverse concentrations of Swainsonine, cell growth, migration and invasion in U251 and LN444 cells were appraised by the common-used CCK-8, BrdU, flow cytometry and Transwell assays. MiR-92a mimic, inhibitor and the correlative NC were transfected into U251 and LN444 cells, and assessment of miR-92a expression was by utilizing qRT-PCR. Functions of miR-92a in above-mentioned cell biological processes were analyzed again in Swainsonine-treated cells. The momentous proteins of cell cycle, apoptosis and PI3K/AKT/mTOR pathway were ultimately examined by western blot.
RESULTS
Swainsonine significantly hindered cell proliferation through decreasing cell viability, declining the percentage of BrdU cells, down-regulating CyclinD1 and up-regulating p16 expression. Enhancement of percentage of apoptotic cells was presented in Swainsonine-treated cells via activating cleaved-Caspase-3 and cleaved-Caspase-9. Additionally, Swainsonine impeded the abilities of migration and invasion by decreasing MMP-2, MMP-9, Vimentin and E-cadherin. Repression of miR-92a was observed in Swainsonine-treated cells, and miR-92a overexpression overturned the anti-tumor activity of Swainsonine in glioma cells. Finally, western blot assay displayed that Swainsonine hindered PI3K/AKT/mTOR pathway via regulating miR-92a.
CONCLUSIONS
These discoveries corroborated that Swainsonine exerted anti-tumor impacts on glioma cells via repression of miR-92a, and inactivation of PI3K/AKT/mTOR signaling pathway.
背景
苦马豆素是一种天然的吲哚里西啶生物碱，其在多种癌症中的抗肿瘤活性已被广泛报道。本研究旨在探讨苦马豆素是否对神经胶质瘤细胞具有抗肿瘤作用，并揭示相关的分子机制。
方法
用不同浓度的苦马豆素处理 U251 和 LN444 细胞后，通过 CCK-8、BrdU、流式细胞术和 Transwell 实验评估细胞生长、迁移和侵袭。将 miR-92a 模拟物、抑制剂和相应的对照物转染到 U251 和 LN444 细胞中，通过 qRT-PCR 评估 miR-92a 的表达。在苦马豆素处理的细胞中再次分析 miR-92a 在上述细胞生物学过程中的功能。最后，通过 Western blot 检测细胞周期、凋亡和 PI3K/AKT/mTOR 通路的关键蛋白。
结果
苦马豆素通过降低细胞活力、降低 BrdU 细胞的百分比、下调 CyclinD1 和上调 p16 表达，显著抑制细胞增殖。在苦马豆素处理的细胞中，通过激活 cleaved-Caspase-3 和 cleaved-Caspase-9 ，增加了凋亡细胞的百分比。此外，苦马豆素通过降低 MMP-2、MMP-9、波形蛋白和 E-钙黏蛋白的表达，抑制了迁移和侵袭的能力。在苦马豆素处理的细胞中观察到 miR-92a 的抑制，而 miR-92a 的过表达推翻了苦马豆素在神经胶质瘤细胞中的抗肿瘤活性。最后，Western blot 实验表明，苦马豆素通过调节 miR-92a 抑制了 PI3K/AKT/mTOR 通路。
结论
这些发现证实，苦马豆素通过抑制 miR-92a 并抑制 PI3K/AKT/mTOR 信号通路对神经胶质瘤细胞发挥抗肿瘤作用。
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