Hsu Ping-Chih, You Bin, Yang Yi-Lin, Zhang Wen-Qian, Wang Yu-Cheng, Xu Zhidong, Dai Yuyuan, Liu Shu, Yang Cheng-Ta, Li Hui, Hu Bin, Jablons David M, You Liang
Thoracic Oncology Laboratory, Department of Surgery, Comprehensive Cancer Center, University of California, San Francisco, CA, USA.
Department of Thoracic Medicine, Chang Gung Memorial Hospital, Linkou, Taoyuan, Taiwan.
Oncotarget. 2016 Aug 9;7(32):51922-51933. doi: 10.18632/oncotarget.10458.
Yes-associated protein (YAP) is a main mediator of the Hippo pathway, which promotes cancer development. Here we show that YAP promotes resistance to erlotinib in human non-small cell lung cancer (NSCLC) cells. We found that forced YAP overexpression through YAP plasmid transfection promotes erlotinib resistance in HCC827 (exon 19 deletion) cells. In YAP plasmid-transfected HCC827 cells, GTIIC reporter activity and Hippo downstream gene expression of AREG and CTGF increased significantly (P<0.05), as did ERBB3 mRNA expression (P<0.05). GTIIC reporter activity, ERBB3 protein and mRNA expression all increased in HCC827 erlotinib-resistance (ER) cells compared to parental HCC827 cells. Inhibition of YAP by small interfering RNA (siRNA) increased the cytotoxicity of erlotinib to H1975 (L858R+T790M) cells. In YAP siRNA-transfected H1975 cells, GTIIC reporter activity and downstream gene expression of AREG and CTGF decreased significantly (P<0.05). Verteporfin, YAP inhibitor had an effect similar to that of YAP siRNA; it increased sensitivity of H1975 cells to erlotinib and in combination with erlotinib, synergistically reduced migration, invasion and tumor sphere formation abilities in H1975 cells. Our results indicate that YAP promotes erlotinib resistance in the erlotinib-sensitive NSCLC cell line HCC827. Inhibition of YAP by siRNA increases sensitivity of erlotinib-resistant NSCLC cell line H1975 to erlotinib.
Yes相关蛋白（YAP）是促进癌症发展的Hippo信号通路的主要调节因子。在此我们表明，YAP可促进人非小细胞肺癌（NSCLC）细胞对厄洛替尼的耐药性。我们发现，通过YAP质粒转染强制YAP过表达可促进HCC827（外显子19缺失）细胞对厄洛替尼产生耐药性。在YAP质粒转染的HCC827细胞中，GTIIC报告基因活性以及AREG和CTGF的Hippo下游基因表达显著增加（P<0.05），ERBB3 mRNA表达也显著增加（P<0.05）。与亲本HCC827细胞相比，HCC827厄洛替尼耐药（ER）细胞中的GTIIC报告基因活性、ERBB3蛋白和mRNA表达均增加。用小干扰RNA（siRNA）抑制YAP可增加厄洛替尼对H1975（L858R+T790M）细胞的细胞毒性。在YAP siRNA转染的H1975细胞中，GTIIC报告基因活性以及AREG和CTGF的下游基因表达显著降低（P<0.05）。YAP抑制剂维替泊芬具有与YAP siRNA类似的作用；它增加了H1975细胞对厄洛替尼的敏感性，并且与厄洛替尼联合使用时，可协同降低H1975细胞的迁移、侵袭和肿瘤球形成能力。我们的结果表明，YAP可促进对厄洛替尼敏感的NSCLC细胞系HCC827对厄洛替尼产生耐药性。用siRNA抑制YAP可增加对厄洛替尼耐药的NSCLC细胞系H1975对厄洛替尼的敏感性。
