Effects of Mindbomb homoIogue 2 gene transfection on proIiferation of human gIioma U251 ceIIs

SHAN Xiaosong; LIU Haipeng; ZHENG Kebin; FANG Chuan; DING Yanan

doi:10.3969/j.issn.1002-266X.2016.43.002

Abstract：Objective To observe the effects of Mindbomb homologue 2 (MIB2)gene transfection on the proliferation of human glioma U251 cells,and to explore its mechanism.Methods U251 cells were divided into four groups:the con-trol group,negative control group,experimental group 1 and experimental group 2.U251 cells in the control group was un-treated;U251 cells in the negative control group was transfected with vector FLAG plasmid;and U251 cells in the experi-mental group 1 was transfected with MIB2-FLAG plasmid;U251 cells in the experimental group 2 was transfected siMIB2. MIB2 mRNA was detected by PCR,the expression of MIB2,nuclear factor-κB (NF-κB)inhibitory protein (IκB)and NF-κB protein was detected by Western blotting in each group,and the optical density (OD)values were detected by MTT.Results The expression of MIB2 mRNA in the experimental group 1,experimental group 2 and negative control group was 19.43 ±3.21,0.43 ±0.17,and 1.03 ±0.12,respectively;and significant difference was found in the MIB2 mRNA between the experimental groups 1,2 and the negative control group (all P <0.01).The expression of MIB2,IκB and NF-κB protein in the experimental group 1 was 5.11 ±0.41,0.49 ±0.13 and 4.99 ±0.32,respectively;the expression of MIB2,IκB and NF-κB protein in the experimental group 2 was 0.43 ±0.17,3.12 ±0.36 and 0.49 ±0.14,respectively;and the expression of MIB2,IκB and NF-κB protein in the negative control group was 1.09 ±0.11,1.02 ±0.46 and 1.01 ± 0.21,respectively.Significant difference was found in the expression of MIB2,IκB and NF-κB protein between the experi-mental groups 1,2 and the negative control group (all P <0.01).At 36,48 and 72 h,compared with the negative control group and blank control group,OD value of U251 cells in the experimental group 1 increased significantly (all P <0.01). Conclusion After MIB2 gene tranfection,the proliferation of human glioma U251 cells increases and MIB 2 gene may play an important role in the regulation of NF-κB signaling pathway through MIB2 protein.