CRISPR/Cas9 gene editing in Huh7 and Hepa RG cell lines

Objectives. Mutations and overexpression of beta-catenin are associated with many cancers including hepatocellular carcinomas (HCC). Activation of beta-catenin signaling may result in resistance to chemotherapeutic agents. The aim of this study was to generate, by CRISPR/Cas9 gene editing technology, a viable cellular model usable to study the detailed mechanisms of HCC and to find target molecules for the development of novel therapeutic drugs. Methods and results. Human Huh7 and HepaRG cells were transfected with CRISPR/CAS beta-catenin KO plasmid (h) plus beta-catenin HDR plasmid (h). Edited clones were validated by fluorescent microscopy and Western Blot analysis and were further cultured. Additionally, the differential response of parental and KO cells to antitumoral drugs was tested. To our knowledge, this is the first report using CRISPR/Cas9 technology on human HCC cell lines to evaluate the correlation between beta-catenin expression and antitumoral drug effects. Conclusion. Our results suggest that beta-catenin is possibly involved in chemotherapy resistance, since the Huh7 beta-catenin(KO) cells appeared to be more sensitive compared to Huh7(WT).