Zhang Lingmin, Li Ying, Chen Qinghua, Xia Yong, Zheng Wenfu, Jiang Xingyu
CAS Center for Excellence in Nanoscience, Beijing Engineering Research Center for BioNanotechnology, CAS Key Laboratory for Biological Effects of Nanomaterials and Nanosafety, National Center for NanoScience and Technology, Beijing 100190, China; Guangdong Provincial Key Laboratory of Molecular Target & Clinical Pharmacology, School of Pharmaceutical Sciences and the Third & Fifth Affiliated Hospital, Guangzhou Medical University, Guangzhou 511436, China.
CAS Center for Excellence in Nanoscience, Beijing Engineering Research Center for BioNanotechnology, CAS Key Laboratory for Biological Effects of Nanomaterials and Nanosafety, National Center for NanoScience and Technology, Beijing 100190, China.
Sci Bull (Beijing). 2018 Nov 15;63(21):1411-1419. doi: 10.1016/j.scib.2018.09.024. Epub 2018 Sep 29.
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 A375 (melanoma) cell line was resistant to inhibitors of B-Raf proto-oncogene, serine/threonine kinase (BRAF), whereas the resultant MED12 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 A375 cells, and a few combinations of EGFR inhibitors and TGF-βR inhibitors were active in suppressing the growth of MED12 PC9 cells. The drug-resistant models will be useful in screening novel drugs and drug combinations for multi-drug-resistant cancer therapy.
癌症治疗常常受到耐药性迅速出现的阻碍。耐药细胞模型对于理解耐药性和开发新疗法至关重要。创建这些模型的低效率和所需的长时间是阻碍耐药性研究和药物筛选的主要障碍。在此，我们报告一种方法，该方法可以通过使用CRISPR/Cas9系统特异性敲除MED12基因来加速（将时间从数年缩短至3周）基于癌细胞系的可遗传耐药模型的建立。所得的MED12 A375（黑色素瘤）细胞系对B-Raf原癌基因丝氨酸/苏氨酸激酶（BRAF）抑制剂具有抗性，而所得的MED12 PC9（非小细胞肺癌）细胞系对表皮生长因子受体（EGFR）抑制剂具有抗性。抗癌药物及其组合的评估表明，BRAF抑制剂和转化生长因子-β受体（TGF-βR）抑制剂的某些组合在抑制MED12 A375细胞生长方面具有活性，而EGFR抑制剂和TGF-βR抑制剂的一些组合在抑制MED12 PC9细胞生长方面具有活性。这些耐药模型将有助于筛选用于多药耐药癌症治疗的新型药物和药物组合。
