Protocol to screen for Sorafenib resistance regulators using pooled lentiviral shRNA library and a Sorafenib-resistant hepatocellular carcinoma cell model

Ruize Gao; Fengyuan Tang; Gerhard Christofori

doi:10.1016/j.xpro.2023.102273

Protocol to screen for Sorafenib resistance regulators using pooled lentiviral shRNA library and a Sorafenib-resistant hepatocellular carcinoma cell model

STAR Protoc. 2023 Apr 30;4(2):102273. doi: 10.1016/j.xpro.2023.102273. Online ahead of print.

Authors

Ruize Gao¹, Fengyuan Tang², Gerhard Christofori²

Affiliations

¹ Department of Biomedicine, University of Basel, CH4058 Basel, Switzerland. Electronic address: ruize.gao@lih.lu.
² Department of Biomedicine, University of Basel, CH4058 Basel, Switzerland.

Abstract

Approaches to study therapy resistance in HCC (hepatocellular carcinoma) are limited, especially when using HCC models in vitro. Here, we present a protocol to establish an in vitro Sorafenib-resistant human HCC cell model and conduct an shRNA-mediated synthetic lethal screen in established Sorafenib-resistant HCC cell lines to identify critical regulators of Sorafenib resistance. We describe steps for RNA sequencing and functional analysis to reveal the mode of action of potential candidates in conferring therapy resistance to HCC cells. For complete details on the use and execution of this protocol, please refer to Gao et al. (2021a)¹ and Gao et al. (2021b).².

Keywords: Cancer; Cell Biology; High Throughput Screening; Molecular Biology; RNAseq; Sequencing.