抗失巢凋亡对胶质瘤细胞增殖和生存的影响

Effects of anoikis resistance on glioma cell proliferation and survival

目的抗失巢凋亡与肿瘤侵袭扩散密切相关,本研究主要揭示抗失巢凋亡对U87胶质瘤细胞增殖、生存能力的影响及机制,对了解脑胶质瘤转移有一定意义。方法本研究构建了稳定表达EGFP(增强绿色荧光蛋白)的人脑胶质瘤U87细胞,并利用EGPF表达监控悬浮培养条件下细胞聚集情况。通过CCK-8细胞增殖检测、平板克隆形成检测、流式细胞周期检验及荧光定量PCR技术,研究抗失巢凋亡对U87细胞的增殖、生存的影响及其机制。结果抗失巢凋亡细胞的增殖能力显著增加,单克隆形成能力增强,G1期细胞所占百分比显著减少,而S期细胞百分比显著增加,并且抗失巢凋亡U87细胞中OCT4、SOX1、CCND等基因表达上调(P<0.05)。结论失巢凋亡的胶质瘤细胞对生存环境依赖性较弱,具有更强的增殖和自我更新能力。

Objective Anoikis resistance is closely associated with tumor invasion and metastasis. This study aimed at determining the effects of anoikis resistance on the proliferation and survival of U87 glioma cells and revealing their mechanisms in order to contribute to the understanding of glioma metastasis. Methods The enhanced green fluorescent protein-expressing human glioma cell line U87 was established to monitor the formation of cell aggregates in suspension culture. CCK-8 proliferation assay, colony formation assay, flow cytometry analysis of cell cycle, and real-time PCR were used to determine the effects of anoikis resistance on the proliferation and survival of U87 ceils. Results The anoikis-resistant U87 ceils had significantly increased cell proliferation, enhanced colony formation ability, significantly reduced percentage of cells in G1 phase, and significantly increased percentage of cells in S phase. Moreover, the stem cell-related genes OCT4 and SOX1 and proto-oncogene CCND1 were up-regulated in anoikis-resistant U87 cells （P 〈 0. 05 ）. Conclusions Anoikis-resistant glioma cells are less dependent on survival environment with more robust proliferation and self-renewal capacity.