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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 nuclear factor-κB1 (NF-κB1) in human glioma and glioma cell lines U87 and SHG44 and its relationship with apoptosis and the mechanism of action of NF-κB1 on glioma . Methods Real-time quantitative PCR (qRT-PCR) was used to detect the expression of NF-κB 1 in non-tumor brain tissues and in different grades of human glioma tissues. The above-mentioned cell lines were transfected with NF-κB1 overexpression / silence expression plasmid (NF-κB1 shRNA) and BCL2 silencing expression plasmid (BCL2 shRNA) by liposome method. The effect of NF-κB1 on the apoptosis of glioma cell line U87 and SHG44 was detected by flow cytometry. The expressions of NF-κB1 and BCL2 protein were detected by Western blotting. Results qRT-PCR showed that the expression of NF-κB1 in human glioma was significantly higher than that in non-tumor brain tissue, and the expression level of NF-κB gradually increased with the malignant degree of tumor (all P <0.05). In the same human glioma specimens BCL2 expression level and NF-κB1 expression trend. Flow cytometry results showed that the inhibition of NF-κB1 expression significantly promoted the apoptosis of human glioma cells U87 and SHG44 (all P <0.05). Western blot analysis confirmed that the expression of BCL2 protein was also decreased after the inhibition of NF-κB1 protein expression. Conclusion NF-κB1 can inhibit the early apoptosis of glioma cells by promoting the expression of BCL2.