Ursolic acid promotes apoptosis and mediates transcriptional suppression of CT45A2 gene expression in non-small-cell lung carcinoma harbouring EGFR T790M mutations
Ursolic acid promotes apoptosis and mediates transcriptional suppression of CT45A2 gene expression in non-small-cell lung carcinoma harbouring EGFR T790M mutations
复制标题
熊果酸在携带 EGFR T790M 的 NSCLC 中促进细胞凋亡并介导 CT45A2 基因表达的转录抑制
DOI：
10.1111/bph.14793
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发表时间：
2019-12-26
影响因子：
7.3
通讯作者：
Yin, Wu
中科院分区：
文献类型：
--
作者：
Yang, Kaiyong;Chen, Yan;Yin, Wu
背景和目的在非小细胞肺癌（NSCLC）患者中，上皮生长因子受体（EGFR）的L 858 R/T790 M突变是EGFR TKI治疗获得性耐药的主要原因，限制了其治疗效果。因此，鉴定能够优先杀死携带L 858 R/T790 M突变的NSCLC的药物至关重要。在这里，我们已经评估了熊果酸，从草药来源中分离的活性成分，对厄洛替尼耐药H1975细胞，港口L 858 R/T790 M突变的影响。实验方法应用基因表达综合谱（GEO）分析检测EGFR突变的NSCLC细胞中差异表达的基因。采用AnnexinV-FITC/PI、TUNEL染色、MTT、伤口愈合、RT-PCR、qRT-PCR、western blot、免疫组化、双荧光素酶报告基因和ChIP-PCR等方法研究熊果酸的体内外作用。关键结果癌/睾丸抗原家族45成员A2（CT 45 A2）在H1975细胞中高度表达。CT 45 A2在H1975细胞中的异位表达在体外增加细胞增殖和运动。沉默CT 45 A2表达强烈减弱了H1975细胞的运动性和生长。熊果酸的抗肿瘤作用主要依赖于H1975细胞中CT 45 A2的表达。熊果酸抑制转录因子TCF 4和β-连环蛋白信号介导的CT 45 A2基因转录。结论与意义CT 45 A2是一个新的癌基因与NSCLC的EGFR T790突变。熊果酸通过负性调节β-catenin/TCF 4/CT 45 A2信号通路诱导H1975细胞凋亡并抑制其增殖。因此，熊果酸可能是携带EGFR-L 858 R/T790 M突变的NSCLC的潜在候选治疗。
Background and Purpose In non-small-cell lung carcinoma (NSCLC) patients, the L858R/T790M mutation of the epithelial growth factor receptor (EGFR) is a major cause of acquired resistance to EGFR-TKIs treatment that limits their therapeutic efficacy. Identification of drugs that can preferentially kill the NSCLC harbouring L858R/T790M mutation is therefore critical. Here, we have evaluated the effects of ursolic acid, an active component isolated from herbal sources, on erlotinib-resistant H1975 cells that harbour the L858R/T790M mutation. Experimental Approach Gene expression omnibus (GEO) profiles analyses was applied to detect differentially expressed genes in NSCLC cells harbouring EGFR mutation. AnnexinV-FITC/PI, TUNEL staining, MTT, wound healing, RT-PCR, qRT-PCR, western blots, immunostaining, dual-luciferase reporters and ChIP-PCR were utilized to investigate the effects of ursolic acid in vitro and in vivo. Key Results The cancer/testis antigen family 45 member A2 (CT45A2) was highly expressed in H1975 cells. Ectopic expression of CT45A2 in H1975 cells increased cell proliferation and motility in vitro. Silencing the CT45A2 expression strongly attenuated H1975 cells motility and growth. The anti-cancer effect of ursolic acid was critically dependent on CT45A2 expression in H1975 cells. Ursolic acid suppressed CT45A2 gene transcription mediated by transcriptional factor TCF4 and beta-catenin signalling. Conclusions and Implications CT45A2 is a novel oncogene for NSCLC with an EGFR T790 mutation. Ursolic acid induced apoptosis and inhibited proliferation of H1975 cells by negatively regulating the beta-catenin/TCF4/CT45A2 signalling pathway. Therefore, ursolic acid may be a potential candidate treatment for NSCLC harbouring the EGFR-L858R/T790M mutation.