Knockout of FosB gene changes drug sensitivity and invasion activity via the regulation of Bcl-2, E-cadherin, fi-catenin, and vimentin expression

Our previous research suggested the presence of a novel SETDB1-mediated FosB pathway that could be responsible for the regulation of cell proliferation and invasiveness during anticancer treatments. In this study, we prepared FosB knock-out (FosB-KO) A549 human lung cancer cells using the CRISPR/Cas9 system and examined the physiological and molecular changes that caused. Annexin V and TUNEL assays showed that FosB-KO clones were less sensitive to doxorubicin treatment compared to the control A549 cells. Bcl2 expression and mitochondrial membrane potential were also both markedly increased in FosB-KO clones, which suggests the involvement of Bcl2 in the doxorubicin mediated increase in cell viability demonstrated the FosB-KO clones. Moreover, we identified changes in the migration and transforming activities of the FosB-KO clones that coincided with changes in the expression levels of E- cadherin, fi-catenin, and Vimentin. RT-PCR and qPCR analysis showed that the expressions of Bcl2, E- cadherin, fi-catenin, and Vimentin were regulated at the transcriptional level. Importantly, FosB over-expression in FosB-KO clones restored the expression of Bcl2, Akt, E-cad, fi-catenin, and Vimentin, suggesting that those proteins were tightly regulated by FosB. These data suggest that the FosB gene critically regulates both drug sensitivity and invasion related genes, and does so in a manner coordinated with the function of SETDB1. Therefore, we propose that the FosB gene regulates both drug sensitivity and invasion activity related genes, and also shows coordinated function with SETDB1 for the regulation of target proteins. (c) 2021 Elsevier Inc. All rights reserved.

Automated summary

Our study indicates that the FosB gene plays a critical role in regulating drug sensitivity and invasion-related genes, affecting the expression of Bcl2, E-cadherin, fi-catenin, and Vimentin at the transcriptional level, and shows coordinated function with SETDB1.