EGF和AG1478对GL15细胞增殖、侵袭和GFAP表达的影响

Effect of EGF and AG1478 on the proliferation, invasion and GFAP expression of GL15 cell line

目的探讨表皮生长因子受体(EGFR)信号通路对人胶质母细胞瘤GL15细胞系细胞生物学特性影响的分子机制。方法Gimsa染色分析GL15细胞核型,甲基四唑兰(MTT)和Transwell分别检测表皮生长因子(EGF),AG1478对GL15细胞增殖和侵袭力的影响,逆转录酶-多聚酶链反应(RT-PCR)和Western blot分别检测EGF和AG1478作用后GL15细胞胶质纤维酸性蛋白(GFAP)mRNA和蛋白表达。结果Gimsa染色表明EGFR所在的7号染色体过度扩增。外源性的EGF双向调节GL15细胞增生和生长,小剂量(50ng/ml,100ng/ml)EGF能促进细胞增生和提高细胞侵袭能力以及GFAPmRNA和蛋白表达,而大剂量(200ng/ml)则能降低其增殖和侵袭能力,AG1478则能明显拮抗EGF活性。结论GL15胶质母细胞瘤细胞系的生物学活性与EGFR所在的7号染色体过度扩增密切相关。EGF可双向调节GL15细胞系的生长,AG1478能不完全抑制EGFR的活性。EGFR是基因治疗胶质母细胞瘤的理想靶点。

Objective To investigate the molecular mechanism of epidermal growth factor receptor （EGFR） signal pathway affecting the biology characteritics of glioblastoma cell lines GL15. Methods Tne glioblastoma cell hnes GL15 were cultured, and the karyotype of GL15 was analyzed with Gimsa stain. Tne GL15 proliferation and invasive ability were detected with methyl thiazolyl tetrazolium （M＇IT） and Transwell, and the expression of glial fibrillary acidic protein （GFAP） mRNA and protein were detected with reverse transcriptase-polymerase chain reaction （RT-PCR） and Western blot after epidermal growth factor （EGF） and AG1478 intervention respectively. Results No. 7 chromosome where EGFR located demonstrated over-amplificatitm with Gimsa stain. Estrogenic EGF can regulate the proliferation and invasive ability and the small dose of EGF （50 ng/ml, 100 ng/ml） can promote the proliferation and enhance the invasive ability, but large dose （200 ng/ml ） would decrease that of EGF. AG1478 could inhibit the biological activity of EGF. Conclusion The biological characteristics of glioblastoma cell hnes GL15 are closely related to the over-amplification of No. 7 chromosome where the chromosome of EGFR is located. EGF regulates the growth of GL15 in two-way and AG1478 can inhibit the biological activity of EGFR non-completely. EGFR may be the ideal therapeutic target of gene methods.