阻断NHE1抑制人胶质母细胞瘤细胞增殖和侵袭的研究

doi:10.3877/cma.j.issn.2095-9141.2020.01.010

中华神经创伤外科电子杂志 ›› 2020, Vol. 06 ›› Issue (01) : 39 -43. doi: 10.3877/cma.j.issn.2095-9141.2020.01.010

所属专题：文献；

基础研究

阻断NHE1抑制人胶质母细胞瘤细胞增殖和侵袭的研究

李玉辉¹, 赵喜庆¹, 陈思¹, 刘岩², 刘艳坤³, 李玉凤³^,^†(

)

^1. 063001　唐山市人民医院神经外科
^2. 063210　唐山，华北理工大学生命科学院生物工程系
^3. 063001　唐山市人民医院肿瘤研究所

收稿日期:2019-12-12 出版日期:2020-02-15
通信作者: 李玉凤
基金资助:
唐山市科技创新团队培养计划项目基金(18130203B)

Blockade of NHE1 inhibits the proliferation and invasion of human glioblastoma

Yuhui Li¹, Xiqing Zhao¹, Si Chen¹, Yan Liu², Yankun Liu³, Yufeng Li³^,^†()

^1. Department of Neurosurgery, Tangshan People’s Hospital, Tangshan 063001, China
^2. Department of Bioengineering, College of Life Science, North China University of Science and Technology, Tangshan 063210, China
^3. Cancer Institute, Tangshan People’s Hospital, Tangshan 063001, China

Received:2019-12-12 Published:2020-02-15
Corresponding author: Yufeng Li
About author:
Corresponding author: Li Yufeng, Email: yufeng_li@tsrmyy.cn

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目的

分析Na⁺/H⁺交换蛋白1（NHE1）抑制剂对癌基因BRAF野生型（BRAF^WT）和激活型BRAF^V600E突变的胶质母细胞瘤（GBM）细胞生长和侵袭能力的影响。

方法

NHE1抑制剂Cariporide分别处理U251（BRAF^WT）和AM38（BRAF^V600E）GBM细胞系，乙酰甲酯化的2’,7’-双（2-羧乙基）-5（6）-羧荧光素荧光探针处理细胞并采用紫外分光光度计检测细胞在440 nm与490 nm的荧光强度，计算荧光强度比值以反映NHE1的活性，MTT法检测细胞增殖活性，基质胶-Transwell实验检测细胞侵袭能力。

结果

AM38细胞的NHE1活性、增殖和侵袭能力均显著高于U251细胞，差异均有统计学意义（P=0.006、0.010、0.047）；Cariporide处理的U251和AM38细胞的NHE1活性、增殖和侵袭能力均显著低于溶剂二甲基亚砜处理的U251和AM38细胞，差异均有统计学意义（U251：P=0.012、0.023、0.044；AM38：P=0.006、0.001、0.038）。

结论

采用Cariporide阻断NHE1活性可有效抑制BRAF^WT和BRAF^V600E突变型GBM细胞的增殖和侵袭。

关键词: 胶质母细胞瘤, NHE1抑制剂, BRAF野生型, BRAF^V600E突变型, 增殖, 侵袭

Objective

To analyze the effection of Na⁺/H⁺ exchanger 1(NHE1) on proliferation and invasion abilities of oncogene BRAF wild type(BRAF^WT) and BRAF^V600E mutation type glioblastoma (GBM) cells.

Methods

The U251 (BRAF^WT) and AM38 (BRAF^V600E) GBM cells were treated with NHE1 inhibitor Cariporide, respectively. The cells were treated with acetoxymethyl ester of 2’,7’-Bis (2-carboxyethyl)-5(6)-carboxyfluorescein fluorescent probe and the fluorescence intensities at 440 nm and 490 nm were detected by ultraviolet spectrophotometer. The fluorescence intensity ratio was calculated to reflect NHE1 activity. The proliferative activity was detected by MTT and the invasion ability was detected by matrigel-Transwell assay.

Results

The NHE1 activity, proliferation and invasion abilities of AM38 cells were all significantly higher than those of U251 cells (P=0.006, 0.010, 0.047, respectively). The NHE1 activity, proliferation and invasion abilities of both Cariporide-treated U251 and Cariporide-treated AM38 cells were all markedly decreased compared with those of DMSO-treated U251 and DMSO-treated AM38 cells, respectively (U251: P=0.012, 0.023, 0.044, respectively; AM38: P=0.006, 0.001, 0.038, respectively).

Conclusion

Cariporide could effectively inhibit the proliferation and invasion abilities of BRAF^WT and BRAF^V600E mutation type GBM cells.

Key words: Glioblastoma, Na⁺/H⁺ exchanger 1 inhibitor, BRAF^WT, BRAF^V600E mutation, Proliferation, Invasion

图1 BCECF-AM荧光探针分析Cariporide对U251细胞和AM38细胞内NHE1活性的影响

图2 MTT法分析Cariporide对U251细胞和AM38细胞生长活性的影响

图3 基质胶-Transwell实验分析Cariporide对U251细胞和AM38细胞侵袭能力的影响

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