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Archive > Volume 24 Issue 2 > 2017,24(2):122-126. DOI:10.3872/j.issn.1007-385X.2017.02.004 Prev Next
Basic Research
block cell cycle of non-small cell cancer H1975 cell through AKT-mTOR pathway
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Abstract:
Objective:To study effect of bridging intergrator 1(Bin 1) gene over-expression on cell cycle of non-small cell lung cancer H1975 line cell, and its mechanism. Methods: CMV-MCS-GFP-SV40-Neomucin-Bin1 plasmid carrying Bin1 gene was constructed and was transfected into the H1975 line cell (Bin1+ group). The H1975 line cell (Bin1- group) transfected with empty plasmid and the H1975 line cell (Control group) were set up. Expressions of Bin1 mRNA and protein in the H1975 cell of the 3 groups were respectively detected by RT-PCR and Western blotting assays. Cell cycle changes of the H1975 cell in the various treatment groups were examined by flow cytometry assay. Phosphorylation level of AKT and mTOR as well as expression of cell cycle related proteins (cyclinD1, CDK4 and Rb) in the various treatment groups were detected by Western blotting. Results: Comparing with the Bin1- and control groups, expressions of Bin1 mRNA and protein in the Bin1+ group were obviously up-regulation (P<0.05) , cell cycle of the H1975 cell in the Bin1+ group was blocked in G1 phase (\[60.53±1.89\]% vs \[46.14±1.56\]% and \[47.33±2.07\]%, P<005), and in the H1975 cell of the Bin1+ group, expressions of p-AKT and p-mTOR were significantly down-regulation (P<0.05) , expressions of AKT and mTOR were not significantly difference (P>0.05) , expressions of cyclinD1 and CDK4 proteins were markedly down-regulation and expression of Rb protein was evidently increased (P<0.05). Conclusion: Over-expression of Bin1 gene in the H195 cell could obviously induce blocking of the cell cycle and its mechanism might be caused by inhibiting AKT-mTOR pathway and its cell cycle related proteins.
Keywords:
non-small lung cancer ; bridging intergrator 1(Bin1) ; cell cycle ; AKT-mTOR
Project Supported:
Project supported by the Young Scientists Foundation of the National Natural Science Foundation of China (No. 81201607), the Natural Science Foundation of Hebei Province(No. H2014206320), and the High Level Talents of Hebei Province（No. A201401040）
