FM19G11体外增敏替莫唑胺对MGMT阳性恶性胶质瘤细胞系的作用机制

基础医学与临床 ›› 2021, Vol. 41 ›› Issue (4): 514-520.

FM19G11体外增敏替莫唑胺对MGMT阳性恶性胶质瘤细胞系的作用机制

方黄毅¹, 尤朝国², 庞晨¹, 章钟鼎¹, 张哲¹, 况统帅¹, 盛汉松^3*

1.温州医科大学第二临床医学院, 浙江温州 325035;
2.耳鼻咽喉科,温州医科大学附属第二医院浙江温州 325027;
3.神经外科,温州医科大学附属第二医院浙江温州 325027

收稿日期:2020-03-01 修回日期:2020-08-24 出版日期:2021-04-05 发布日期:2021-04-05
通讯作者: ^*shs951052@163.com
基金资助:
浙江省科技厅基金 (2016C33SA300055);温州市科技局科研基金(Y20180665)

Mechanism of in vitro sensitization of temozolomide to MGMT⁺ glioma cell line by FM19G11

FANG Huang-yi¹, YOU Chao-guo², PANG Chen¹, ZHANG Zhong-ding¹, ZHANG Zhe¹, KUANG Tong-shuai¹, SHENG Han-song^3*

1. the Second Clinical Medical College of Wenzhou Medical University,Wenzhou 325035;
2. Department of Otolaryngology, the Second Affiliated Hospital of Wenzhou Medical University, Wenzhou 325027, China;
3. Department of Neurosurgery, the Second Affiliated Hospital of Wenzhou Medical University, Wenzhou 325027, China

Received:2020-03-01 Revised:2020-08-24 Online:2021-04-05 Published:2021-04-05
Contact: ^*shs951052@163.com

摘要/Abstract

摘要： 目的观察FM19G11是否影响人源性恶性胶质瘤细胞系T98G对替莫唑胺(TMZ)的敏感性以及相关的分子机制。方法以0、0.5、1和2 μmol/L的FM19G11和TMZ分别处理以及二者联合处理T98G细胞,用CCK-8法和细胞克隆形成实验检测T98G细胞增殖;用Hoechst 33258染色法检测T98G细胞的凋亡形态;用RT-qPCR法和Western blot分别检测T98G细胞HIF-1α、VEGF和EPO的mRNA和蛋白以及O⁶-甲基鸟嘌呤-DNA甲基转移酶(MGMT)和NF-κB P65蛋白的表达。结果与TMZ单独处理相比,FM19G11和TMZ的联合处理对T98G的细胞增殖抑制作用和凋亡形态改变的程度更加明显(P＜0.05); HIF-1α、VEGF和EPO的mRNA和蛋白表达降低(P＜0.05); MGMT、NF-κB P65的蛋白表达降低(P＜0.05)。结论 FM19G11通过靶向抑制HIF-1α下调MGMT蛋白的表达,从而增加T98G细胞对TMZ的药物敏感性。同时FM19G11还能下调HIF-1α下游与肿瘤增殖转移相关的基因并抑制NF-κB信号通路,进一步抑制肿瘤的生长。

关键词: 恶性胶质瘤, FM19G11, 替莫唑胺, 低氧诱导因子-1α, O⁶-甲基鸟嘌呤-DNA甲基转移酶

Abstract: Objective To investigate whether FM19G11 affects the sensitivity of human-derived malignant glioma cell line T98G to temozolomide (TMZ) and underlying molecular mechanisms. Methods T98G cells were treated with 0, 0.5, 1 and 2 μmol/L of FM19G11 and TMZ separately and jointly. The CCK-8 assay and cell clone formation assay were used to detect the proliferation of T98G cells. The Hoechst 33258 staining assay was used to detect the morphological changes of apoptosis in T98G cells. RT-PCR was used to detect the mRNA expression of HIF-1α and its downstream genes VEGF and EPO. Western blot was used to detect the protein level of O⁶-methylguanine-DNA methyltransferase (MGMT), NF-κB P65, HIF-1α,VEGF and EPO. Results Compared with the treatment with TMZ alone, the combined treatment of FM19G11 plus TMZ had a more pronounced effect on the cell proliferation inhibition and morphological changes of apoptosis in T98G cells(P＜0.05). The mRNA and protein of HIF-1α,VEGF and EPO and the protein expression of MGMT, NF-κB P65 were all decreased.(P＜0.05). Conclusions FM19G11 mainly inhibits the expression of HIF-1α, and then down-regulates the expression of MGMT protein, thereby increasing the sensitivity of glioblastoma T98G cells to TMZ. At the same time, FM19G11 also down-regulates genes relating to tumor proliferation and metastasis downstream of HIF-1α and inhibit the NF-κB signaling pathway and then inhibits tumor growth.

Key words: glioblastoma, FM19G11, temozolomide, hypoxia inducible factor 1α, O⁶-methylguanine DNA-methyltransferase

中图分类号:

R739.41

方黄毅, 尤朝国, 庞晨, 章钟鼎, 张哲, 况统帅, 盛汉松. FM19G11体外增敏替莫唑胺对MGMT阳性恶性胶质瘤细胞系的作用机制[J]. 基础医学与临床, 2021, 41(4): 514-520.

FANG Huang-yi, YOU Chao-guo, PANG Chen, ZHANG Zhong-ding, ZHANG Zhe, KUANG Tong-shuai, SHENG Han-song. Mechanism of in vitro sensitization of temozolomide to MGMT⁺ glioma cell line by FM19G11[J]. Basic & Clinical Medicine, 2021, 41(4): 514-520.

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