Effects of AQP5 gene silencing on proliferation, migration and apoptosis of human glioma cells through regulating EGFR/ERK/ p38 MAPK signaling pathway.
Effects of AQP5 gene silencing on proliferation, migration and apoptosis of human glioma cells through regulating EGFR/ERK/ p38 MAPK signaling pathway.
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AQP5基因沉默调控EGFR/ERK/p38 MAPK信号通路对人胶质瘤细胞增殖、迁移和凋亡的影响
DOI：
10.18632/oncotarget.16461
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发表时间：
2017-06-13
期刊：
影响因子：
--
通讯作者：
Zhao CR
中科院分区：
文献类型：
--
作者：
Yang J;Zhang JN;Chen WL;Wang GS;Mao Q;Li SQ;Xiong WH;Lin YY;Ge JW;Li XX;Gu Z;Zhao CR
本研究通过调控EGFR/ERK/p38MAPK信号通路，研究水通道蛋白5(AQP5)基因沉默对人脑胶质瘤细胞增殖、迁移和凋亡的影响。采用定量逆转录聚合酶链式反应(QRT-PCR)检测AQP5在5种人脑胶质瘤细胞系中的表达。选择U87-MG、U251和LN229细胞，随机分为空白组、载体组、AQP5 siRNA组和FlagAQP5组。用四甲基偶氮唑盐比色法检测细胞增殖。AnnexinV-FITC/PI双染流式细胞术(FCM)和PI染色分别检测细胞凋亡和细胞周期。采用划痕试验检测细胞迁移情况。Western blotting检测EGFR/ERK/p38MAPK信号通路相关蛋白。结果表明，AQP5在原发胶质母细胞瘤中的阳性表达与肿瘤大小、是否完全切除有关。U87-MG、U251和LN229细胞株AQP5的mRNA表达显著高于U373和T98G细胞。AQP5 siRNA组U87-MG、U251和LN229细胞的增殖率均低于载体组和空白组。与载体组相比，AQP5 siRNA组细胞凋亡率增加。划痕实验表明，AQP5基因沉默可抑制细胞迁移。与载体组和空白组相比，AQP5 siRNA组ERK1/2、p38MAPK、p-ERK1/2和p-p38MAPK蛋白表达降低。AQP5基因沉默通过抑制EGFR/ERK/p38 MAPK信号通路，抑制U87-MG、U251和LN229细胞的增殖，减少细胞迁移，促进细胞凋亡。
We investigated the effects of aquaporin 5 (AQP5) gene silencing on the proliferation, migration and apoptosis of human glioma cells through regulating the EGFR/ERK/p38MAPK signaling pathway. qRT-PCR was applied to examine the mRNA expressions of AQP5 in five human glioma cell lines. U87-MG, U251 and LN229 cells were selected and assigned into blank, vector, AQP5 siRNA and FlagAQP5 groups. MTT assay was used to measure cell proliferation. Flow cytometry (FCM) with AnnexinV-FITC/PI double staining and PI staining were employed to analyze cell apoptosis and cell cycle respectively. Scratch test was used to detect cell migration. Western blotting was performed to determine the EGFR/ERK/p38 MAPK signaling pathway-related proteins. Results showed that the positive expression of AQP5 in primary glioblastoma was associated with the tumor size and whether complete excision was performed. The mRNA expressions of AQP5 in cell lines of U87-MG, U251 and LN229 were significantly higher than in U373 and T98G. The proliferation rates of U87-MG, U251 and LN229 cells in the AQP5 siRNA group were lower than in the vector and blank groups. The apoptosis rate increased in the AQP5 siRNA group compared with the vector group. Scratch test demonstrated that AQP5 gene silencing could suppress cell migration. Compared with the vector and blank groups, the AQP5 siRNA group showed decreased expressions of the ERK1/2, p38 MAPK, p-ERK1/2 and p-p38 MAPK proteins. AQP5 gene silencing could inhibit the cell proliferation, reduce cell migration and promote the cell apoptosis of U87-MG, U251 and LN229 by suppressing EGFR/ERK/p38 MAPK signaling pathway.