The construction of drug-resistant cancer cell lines by CRISPR/Cas9 system for drug screening

作     者：Lingmin Zhang Ying Li Qinghua Chen Yong Xia Wenfu Zheng Xingyu Jiang 

作者机构：CAS Center for Excellence in NanoscienceBeijing Engineering Research Center for BioNanotechnologyCAS Key Laboratory for Biological Effects of Nanomaterials and NanosafetyNational Center for NanoScience and TechnologyBeijing 100190China University of Chinese Academy of SciencesBeijing 100049China Guangdong Provincial Key Laboratory of Molecular Target &Clinical PharmacologySchool of Pharmaceutical Sciences and the Third &Fifth Affiliated HospitalGuangzhou Medical UniversityGuangzhou 511436China 

出 版 物：《Science Bulletin》 (科学通报（英文版）)

年 卷 期：2018年第63卷第21期

页      面：1411-1419页

核心收录：

学科分类：07[理学] 

基　　金：supported by the Minister of Science and Technology of China (2017YFA0205901) the National Natural Science Foundation of China (21535001, 81730051, 81673039, 31470911) CAS/SAFEA International Partnership Program for Creative Research Teams 

主　　题：CRISPR/Cas9 Drug resistance Cancer Cell line Gene editing 

摘      要：Cancer therapy is often hampered by the rapid emergence of drug resistance. Drug-resistant cellular models are essential for understanding the drug resistance and developing new therapeutics. The low efficiency and long time required in creating these models are major obstacles hindering drug resistance research and drug screening. Herein, we report an approach that can accelerate(shortening the time from years to 3 weeks) the establishment of cancer cell line-based, inheritable drug resistance models by specific knockout of MED12 gene using CRISPR/Cas9 system. The resultant MED12^(KO) A375(melanoma)cell line was resistant to inhibitors of B-Raf proto-oncogene, serine/threonine kinase(BRAF), whereas the resultant MED12^(KO) PC9(non-small cell lung cancer) cell line was resistant to inhibitors of epidermal growth factor receptor(EGFR). Evaluation of anti-cancer drugs and their combinations shows that certain combinations of BRAF inhibitors and TGF-β receptor(TGF-βR) inhibitors are active in suppressing the growth of MED12^(KO) A375 cells, and a few combinations of EGFR inhibitors and TGF-βR inhibitors were active in suppressing the growth of MED12^(KO) PC9 cells. The drug-resistant models will be useful in screening novel drugs and drug combinations for multi-drug-resistant cancer therapy.