桥接整合因子1通过AKT-mTOR通路诱导非小细胞肺癌H1975细胞周期阻滞

发布时间：2018-06-06 08:31

本文选题：非小细胞肺癌 + 桥接整合因子　；参考：《中国肿瘤生物治疗杂志》2017年02期

【摘要】：目的:研究桥接整合因子1(bridging intergrator 1,Bin1)基因过表达后对非小细胞肺癌细胞株H1975细胞周期的影响及其作用机制。方法:构建携带Bin1基因的CMV-MCS-GFP-SV40-Neomycin-Bin1质粒,并转染H1975细胞(Bin1~+组),另设置空白质粒转染组(Bin1~-组)及空白对照组(Ctrl组),利用RT-PCR和Western blotting分别检测3组细胞中Bin1在mRNA和蛋白质水平的表达情况。流式细胞术检测不同处理组H1975细胞周期的变化,Western boltting分别检测各组中AKT、mTOR磷酸化水平及细胞周期相关蛋白(周期蛋白D1、CDK4、Rb)的表达情况。结果:与Bin1~-组、Ctrl组比较,Bin1~+组H1975细胞中Bin1在mRNA、蛋白水平表达明显上调(均P0.05);H1975细胞阻滞在G1期[(60.53±1.89)%vs(46.14±1.56)%、(47.33±2.07)%,均P0.05];Bin1~+组H1975细胞内p-AKT、p-mTOR表达下调(均P0.05),AKT、mTOR表达变化无统计学差异(P0.05);周期蛋白D1、CDK4的表达量均明显下调(P0.05),Rb表达量明显增加(P0.05)。结论:Bin1基因在H1975细胞株过表达后明显诱导细胞周期阻滞,其机制可能是通过抑制AKT-mTOR通路及其细胞周期相关蛋白实现的。
[Abstract]:Aim: to investigate the effect of over-expression of 1(bridging intergrator _ 1 / Bin1 gene on cell cycle of non-small cell lung cancer (NSCLC) cell line H1975 and its mechanism. Methods: the CMV-MCS-GFP-SV40-Neomycin-Bin1 plasmid carrying Bin1 gene was constructed and transfected into H1975 cell line Bin1- group, and the blank plasmid was added to the control group. The expression of Bin1 in mRNA and protein was detected by RT-PCR and Western blotting, respectively. The changes of H1975 cell cycle in different treatment groups were detected by flow cytometry. The phosphorylation level of AKTnmTOR and the expression of cyclin D1- CDK4- Rb were detected by Western boltting. Results: compared with BIN1M-Ctrl group, the expression of Bin1 in H1975 cells in Bin1 ~ group was significantly up-regulated (P0.05 + H1975 cells were blocked in G1 phase [60.53 卤1.89)%vs(46.14 卤1.560.33 卤2.07, P0.05]. There was no significant difference in the expression of p-AKTmTOR in H1975 cells (P 0.05). The expression of CDK4 in D1 was significantly decreased, and the expression of RB in P0. 05 was significantly increased. Conclusion the cell cycle arrest induced by the overexpression of the cell line is probably due to the inhibition of the AKT-mTOR pathway and its cell cycle associated proteins.
【作者单位】：河北医科大学第四医院生物治疗科;
【基金】：国家自然科学基金青年科学基金资助项目(No.81201607) 河北省杰出青年基金资助项目(No.H2014206320) 河北省高层次人才培养项目资助(No.A201401040)~~
【分类号】：R734.2

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