表皮生长因子受体反义基因转染对人恶性胶质瘤细胞体外生长的影响

The effect of antisense epidermal growth factor receptor RNA on the growth of human malignant glioma cells

目的探讨表皮生长因子受体（ＥＧＦＲ）反义基因转染对人恶性胶质瘤细胞Ｕ８７ＭＧ体外生长的影响。方法构建ＥＧＦＲ反义基因表达载体，应用脂质体转染法将其导入Ｕ８７ＭＧ细胞；ＰＣＲ方法鉴定转染克隆；Ｗｅｓｔｅｒｎ印迹杂交检测ＥＧＦＲ蛋白表达水平；通过绘制生长曲线和软琼脂集落形成实验测定反义ＥＧＦＲ基因对恶性胶质瘤细胞体外生长的抑制作用。结果转染反义ＥＧＦＲ基因的细胞克隆株ＡＳ１ＡＳ６的ＥＧＦＲ蛋白表达水平均有不同程度的降低；ＡＳ１ＡＳ６细胞体外生长明显减慢，ＡＳ１ＡＳ６细胞克隆形成率明显降低。表明Ｕ８７ＭＧ细胞体外生长速度和恶性转化能力均与ＥＧＦＲ表达呈正相关。结论反义ＥＧＦＲ基因转染可以明显抑制Ｕ８７ＭＧ细胞生长。

Objective To study the effect of antisense EGFR RNA on the growth of human malignant glioma cells.Methods An antisense EGFR expression vector was constructed and introduced into human malignant glioma cells. PCR analysis was used to determine which clones were successfully transfected with antisense EGFR constructs. Western blotting analysis was used to determine EGFR protein level in the transfected cells. The in vitro growth and their ability to grow in soft agar were studied.Results Western blotting analysis showed six clones stably expressing lower levels of EGFR protein. Antisense EGFR RNA was found to inhibit the proliferation of human malignant glioma cells and their ability to grow in soft agar. There was a negative correlation between the growth inhibiting effect of antisense EGFR and EGFR protein expression. Conclusion These results demonstrate that EGFR play an pivotal role in human malignant glioma cell growth. Antisense EGFR can inhibit the growth of human malignant glioma cells.