STAT1基因对胶质瘤细胞系U251细胞周期的影响及其机制

黄平; 窦长武; 鞠海涛; 王宏伟; 肖瑞; 牛丽丽

doi:10.3971/j.issn.1000-8578.2019.18.1201

STAT1基因对胶质瘤细胞系U251细胞周期的影响及其机制

Effect of STAT1 on Cell Cycle of Human Glioma U251 Cells and Related Mechanism

摘要

摘要:
目的探讨信号转导与转录激活因子1（STAT1）对胶质瘤细胞系U251细胞周期的影响及其机制。
方法利用Lipofectamine^TM2000转染试剂体外瞬时将pcDNA3.1-STAT1转染入胶质瘤细胞系U251，将细胞分为Mock组（无转染）、空载组（转染pcDNA3.1）和STAT1组（转染pcDNA3.1-STAT1），采用Western blot法检测胶质瘤U251细胞中STAT1表达水平，MTT法检测转染STAT1的U251细胞增殖活性，流式细胞仪检测细胞周期指标，划痕实验检测细胞迁移指标，通过Western blot法检测转染细胞中P53、P21、bcl-2、Caspase-8的表达水平及变化趋势。
结果与Mock和空载组相比，STAT1组中STAT1蛋白表达量明显增高（P < 0.05），细胞增殖明显减慢（P < 0.05），G₀/G₁期细胞比例明显升高，细胞的迁移能力明显下降；P53、P21、Caspase-8表达明显增强（P < 0.05），bcl-2表达明显减弱（P < 0.05）。
结论高表达的STAT1对人脑胶质瘤U251细胞具有抑制增殖或促进凋亡的作用，并且STAT1可调控多分子信号转导通路，对脑胶质瘤的发生和发展起到了关键的作用。

Abstract:
Objective To investigate the effect of signal transducer and activator of transcription 1 (STAT1) on cell cycle of glioma U251 cells and related mechanism.
Methods We transiently transfected glioma cell U251 cells with pcDNA3.1-STAT1 using Lipofectamine^TM2000 transfection reagent. The cells were divided into Mock group (no transfection), empty vector group (transfected with pcDNA3.1) and STAT1 group (transfected with pcDNA3.1-STAT1). The expression of STAT1 in glioma U251 cells was detected by Western blot. Cell proliferation was detected by MTT. Cell cycle was detected by flow cytometry. Wound healing assay was used to detect cell migration. The expression of P53, P21, bcl-2 and Caspase-8 in transfected cells was detected by Western blot.
Results Compared with Mock group and empty vector group, the expression of STAT1 protein in STAT1 group was significantly increased (P < 0.05), cell proliferation decreased significantly (P < 0.05), the percentage of cells in phase G₀/G₁ increased significantly, and cell migration ability decreased significantly. The expression levels of P53, P21 and Caspase-8 increased significantly (P < 0.05) while bcl-2 decreased significantly (P < 0.05).
Conclusion The high expression of STAT1 could inhibit the proliferation and promote the apoptosis of human glioma U251 cells, and STAT1 could regulate the expression of multiple signal transduction molecules. STAT1 plays a key role in the occurrence and development of glioma.

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