摘要
目的探讨核转录因子-κB1(NF-κB1)在人脑胶质瘤和胶质瘤细胞株U87、SHG44的表达及其与凋亡的相关性;以及NF-κB1对脑胶质瘤的作用机制。方法利用实时荧光定量PCR(qRT-PCR)检测NF-κB1在非瘤脑组织和不同级别人脑胶质瘤组织中的表达水平。采用脂质体法将化学合成NF-κB1过表达/沉默表达质粒(NF-κB1 shRNA),以及BCL2沉默表达质粒(BCL2 shRNA)转染上述细胞系。通过流式细胞术检测NF-κB1对胶质瘤细胞株U87、SHG44凋亡的影响,并采用Western blotting分别检测NF-κB1、BCL2蛋白的表达水平。结果 qRT-PCR显示,NF-κB1在人脑胶质瘤中表达明显高于非瘤脑组织,并且表达水平随着肿瘤恶性程度的增高而逐渐升高(均P<0.05)。在同一人脑胶质瘤标本中BCL2的表达水平与NF-κB1的表达趋势相同。流式细胞术检测结果显示,抑制NF-κB1表达明显促进人脑胶质瘤细胞U87、SHG44的凋亡(均P<0.05)。Western blot检测证实,抑制NF-κB1蛋白表达后BCL2蛋白的表达水平也随之降低。结论 NF-κB1通过促进BCL2的表达而抑制胶质瘤细胞的早期凋亡。
Objective：To investigate the expression of NF-kappaB1 （NFκ-B1） in human glioma and glioma cell lines SHG44 and U87 and its correlation with apoptosis,which lay a theoretical foundation for the mechanism of NFκ-B1 in human glioma. Methods：qRT-PCR was used to determine the expression of NFκ-B1 in noncancerous brain tissue and human glioma tissue samples respectively.Chemical synthesis of NFκ-B1 silencing expression plasmids was transfected into the cell lines by the Lipofectamine 2000 liposome.The effects of NFκ-B1 on the apoptosis of glioma cell lines SHG44 and U87 were detected by flow cytometry.The expression of NFκ-B1 and BCL2 proteins were detected by western blot. Results：qRT-PCR results showed that the expression of NFκ-B1 in human glioma and glioma cell lines SHG44 and U87 were significantly higher than those in non-brain tissue and increased with the increase of tumor malignancy.The expression of BCL2 was the same in the same human glioma specimen.The results of flow cytometry showed that inhibition of NFκ-B1 expression significantly promoted the apoptosis of SHG44 and U87 in human glioma cells（P〈0.05）.Western blot analysis further confirmed that the expression of BCL2 protein was also decreased after inhibition of NFκ-B1 protein expression. Conclusion：NFκ-B1 inhibits early apoptosis of glioma cells by promoting the expression of BCL2.
出处
《临床神经外科杂志》
CAS
2017年第5期326-330,共5页
Journal of Clinical Neurosurgery
基金
国家自然科学基金(NSFC 81372689)
江苏省卫生和计划生育委员会2016年度青年科研课题(Q201606)
苏州市卫生和计划生育委员会"科教兴卫"项目(kjxw2016020)
