Golgi phosphoprotein 3 sensitizes the tumour suppression effect of gefitinib on gliomas
Golgi phosphoprotein 3 sensitizes the tumour suppression effect of gefitinib on gliomas
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高尔基体磷蛋白3敏化吉非替尼对胶质瘤的抑癌作用
DOI：
10.1111/cpr.12636
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发表时间：
2019-07-01
影响因子：
8.5
通讯作者：
Zhou, Xiuping
中科院分区：
文献类型：
--
作者：
Wang, Xu;Wang, Zhaohao;Zhou, Xiuping
目的研究高尔基体磷蛋白3（Golgi phosphoprotein 3，GOLPH 3）通过抑制EGFR的内吞和降解，导致EGFR的积聚和PI 3 K-AKT通路的过度激活，从而促进胶质瘤的进展。在目前的研究中，我们研究GOLPH 3是否影响胶质瘤细胞对EGFR选择性抑制剂吉非替尼的反应。材料与方法采用免疫荧光和免疫印迹法检测GOLPH 3和EGFR在胶质瘤细胞中的表达。采用CCK-8、EdU掺入法和克隆形成试验测定细胞活力或体外生长。原代胶质瘤细胞用胰蛋白酶和机械消化法培养。transwell侵袭实验检测原代胶质瘤细胞的运动能力。建立裸鼠颅内胶质瘤模型，探讨吉非替尼对GOLPH 3高表达肿瘤细胞的体内敏感性。结果GOLPH 3过表达的永生化胶质瘤细胞和原代胶质瘤细胞均具有较高的EGFR蛋白表达水平，对吉非替尼的敏感性也较高。此外，具有较高GOLPH 3水平的原代胶质瘤细胞表现出更强的增殖行为。重要的是，GOLPH 3增强了吉非替尼的体内抗肿瘤作用。同样，吉非替尼治疗后，GOLPH 3过表达细胞衍生的肿瘤显示出较低的Ki 67阳性和较高的裂解半胱天冬酶-3阳性细胞比对照肿瘤。结论GOLPH 3可增加胶质瘤细胞对吉非替尼的敏感性。本研究为进一步探讨GOLPH 3高表达胶质瘤是否对EGFR治疗更敏感提供了基础，也为开发针对胶质瘤个体化的新的治疗方法提供了思路。
Objectives We previously reported that Golgi phosphoprotein 3 (GOLPH3) promotes glioma progression by inhibiting EGFR endocytosis and degradation, leading to EGFR accumulation and PI3K-AKT pathway over-activation. In the current study, we examine whether GOLPH3 affects the response of glioma cells to gefitinib, an EGFR selective inhibitor. Materials and Methods The expression of GOLPH3 and EGFR in glioma cells was detected by immunofluorescence and immunoblotting. The cell viability or growth in vitro was determined by CCK-8, EdU incorporation and clonogenic assays. The primary glioma cells were cultured by trypsin and mechanical digestion. The transwell invasion assay was used to examine the primary glioma cell motility. Intracranial glioma model in nude mice were established to explore the sensitivity of gefitinib to GOLPH3 high cancer cells in vivo. Results Both the immortalized and primary glioma cells with GOLPH3 over-expression hold higher EGFR protein levels on the cell membrane and exhibited higher sensitivity to gefitinib. In addition, primary glioma cells with higher GOLPH3 level exhibited stronger proliferation behaviour. Importantly, GOLPH3 enhanced the anti-tumour effect of gefitinib in vivo. Consistently, after gefitinib treatment, tumours derived from GOLPH3 over-expression cells exhibited lower Ki67-positive and higher cleaved caspase-3-positive cells than control tumours. Conclusions Our results demonstrate that GOLPH3 increases the sensitivity of glioma cells to gefitinib. Our study provides foundation for further exploring whether GOLPH3 high gliomas will be more sensitive to anti-EGFR therapy in clinic and give ideas for developing new possible treatments for individual glioma patients.