摘要
目的 探讨表皮生长因子促进胶质瘤U251细胞的增殖和侵袭是否涉及电压-门控钠离子通道（VGSCs）亚型nNav1.5的表达.方法 设计合成nNav1.5特异性siRNA,脂质体转染U251细胞,用Western blot法检测转染效率;表皮生长因子（100 ng/L）处理空白组和转染组细胞后,用Real-time PCR和Western blot法分别检测nNav1.5 mRNA和蛋白在各组细胞中的表达水平,并用改良MTT法和Matrigel侵袭实验检测各组细胞增殖及侵袭能力的变化.结果 siRNA成功转染U251细胞;100 ng/L表皮生长因子显著上调U251细胞nNav1.5 mRNA和蛋白的表达的水平（P＜0.05）,并增加肿瘤细胞的增殖和侵袭（P＜0.05）;在siRNA沉默U251细胞nNav1.5表达后,100 ng/L表皮生长因子上调U251细胞nNav1.5 mRNA和蛋白的表达水平能力下降（P＜0.05）,而且促进肿瘤细胞增殖和侵袭的能力明显下降（P＜0.05）.结论 表皮生长因子可通过上调nNav1.5的表达促进胶质瘤U251细胞增殖和侵袭能力.
Objective To investigate whether epidermal growth factor （EGF） promoting the proliferation and invasion of glioma U251 cells was involved in the expression of voltage-gated sodium channels （VGSCs） subtype nNavl. 5. Methods The synthesis of nNavl. 5 specific siRNA was designed and the liposome transfected U251 cells. The transfeetion effieiency was tested by Western blot. After treating the cells of the blank group and the transfection group, real-time PCR and Western blot were used to detect the changes of expression level of nNavl. 5 mRNA and proteins in each group, and the modified MTT method and Matrigel invasion experiment were used to detect the changes of cellular proliferation and invasion ability in each group. Results siRNA were successfully transfected into U251 cells; 100 ng/L EGF significantly upregulated the expression levels of nNavl. 5 mRNA and protein in U251 cells （P 〈 0. 05）, and increased the proliferation and invasion of glioma cells （ P 〈 0.05 ）. After the expression of siRNA silenced U251 cells nNavl. 5, the ability of 100 ng/L EGF upregulating the nNavl. 5 mRNA in U251 cells and protein expression levels was decreased （ P 〈 0.05 ） , and the ability of promoting the proliferation and invasion of glioma cells was decreased obviously （ P 〈 0.05 ）. Conclusion EGF may promote the proliferation and invasion ability of glioma U251 ceils by upregulating the expression of nNavl. 5.
出处
《中华神经外科杂志》
CSCD
北大核心
2016年第8期846-850,共5页
Chinese Journal of Neurosurgery
基金
山东大学第二医院科研基金（S2014010011）
