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目的研究Mind Bomb-2(MIB2)的表达对脑胶质瘤细胞U251增殖的影响。方法用蛋白质印迹法检测正常星形胶质细胞和胶质瘤细胞U251和MIB2蛋白的表达情况。将U251细胞、转染FLAG(一种20个碱基编码的标记蛋白)质粒的U251细胞和转染MIB2-FLAG质粒的U251细胞对应分为空白组、对照组和实验组。以蛋白质印迹法检测上述3组中MIB2的相对含量;采用实时定量PCR技术检测MIB2 mRNA在3组U251细胞中的表达情况;以蛋白质印迹法检测转染后U251细胞中MIB2、I Kappa B(IkB)和核因子κB(NF-κB)的表达;利用四甲基偶氮唑盐(MTT)实验检测转染后U251细胞的增殖情况。结果 U251细胞MIB2的相对含量是星形胶质细胞的(17.04±2.91)倍;经MIB2-FLAG质粒转染U251细胞后,与对照组相比,实验组MIB2 mRNA增加(20.02±2.11)倍,实验组MIB2和NF-κB的蛋白表达增加(6.33±0.32)倍和(5.21±0.21)倍,实验组Ik B的表达量为对照组的(0.43±0.04)倍;实验组MIB2-FLAG质粒转染U251细胞生长增殖速度明显增加(P<0.001)。结论 MIB2通过调节NF-κB信号通路促进U251增殖。
Objective To study the effect of MIB2 expression on the proliferation of glioma U251 cells. Methods Western blotting was used to detect the expression of U251 and MIB2 protein in normal astrocytes and glioma cells. U251 cells, U251 cells transfected with FLAG (a 20-base-encoding marker protein) plasmid and U251 cells transfected with MIB2-FLAG plasmid were divided into blank group, control group and experimental group. The relative content of MIB2 in these three groups was detected by Western blotting. The expression of MIB2 mRNA in U251 cells was detected by real-time quantitative PCR. The expression of MIB2, IκBα, ) And nuclear factor kappa B (NF-κB). The proliferation of U251 cells was detected by MTT assay. Results The relative content of MIB2 in U251 cells was (17.04 ± 2.91) times higher than that in U251 cells. Compared with the control group, the MIB2 mRNA in MIB2-FLAG plasmid transfected U251 cells increased (20.02 ± 2.11) times, The protein expression of MIB2 and NF-κB increased by (6.33 ± 0.32) times and (5.21 ± 0.21) times in the experimental group, and the expression of IκB in the experimental group was (0.43 ± 0.04) times of the control group. The MIB2-FLAG plasmid U251 cells were significantly increased proliferation rate (P <0.001). Conclusion MIB2 promotes U251 proliferation by regulating NF-κB signaling pathway.