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. 2013 Feb 28;8(2):e53072. doi: 10.1371/journal.pone.0053072

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© 2013 Peng et al

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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(A) Construction of the AFP-Cre/LoxP-shRNA system targeting Atg5 gene (AFP-Cre/LoxP-shRNA-Atg5). Q-PCR and western blot analysis showed that it could efficiently knockdown Atg5 gene. (B) Combination of AFP-Cre/LoxP-shRNA-Atg5 infection and sorafenib treatment induced a significantly increase in apoptosis of HCC cells (MHCC97L and PLC) as compared to sorafenib treatment alone. Apoptosis was quantified by annexin V/PI FCM analysis. (C) Q-PCR and western blot analysis showed that intratumoral injection of the AFP-Cre/LoxP-shRNA-Atg5 efficiently silenced Atg5 gene in vivo. (D) The AFP-Cre/LoxP-shRNA-Atg5 significantly enhanced sorafenib-induced suppression of tumorigenicity of MHCC97L cells in vivo. The tumor weight of mice receiving intratumoral injection of the AFP-Cre/LoxP-shRNA-Atg5 combined with sorafenib was significantly lower than that of mice subjected to sorafenib alone. (Cre+LoxP: infection of AFP-Cre/LoxP-shRNA-Atg5; AFP-Cre and LoxP-shRNA: infection of AFP-Cre or LoxP-shRNA alone). (*P<0.05, **P<0.01).