Yin Yunhong, Yang Yie, Yang Liyun, Yang Yan, Li Chunyu, Liu Xiao, Qu Yiqing
Department of Respiratory Medicine, Qilu Hospital of Shandong University, Jinan, 250012, China.
Clinical Laboratory, Qianfoshan Hospital of Shandong Province, Jinan, 250012, China.
Tumour Biol. 2016 Feb;37(2):2597-602. doi: 10.1007/s13277-015-4093-8. Epub 2015 Sep 21.
Lung cancer is the major cause of cancer-related death worldwide, and 80 % of them are non-small cell lung cancer (NSCLC) cases. Gremlin, a bone morphogenetic protein (BMP) antagonist, is overexpressed in various cancerous tissues; however, little is known about the roles of Gremlin in lung carcinogenesis, and it remains unclear whether Gremlin expression may associate with EGFR-TKI resistance. In this study, expression of Gremlin mRNA and protein in matched tumor and normal lung specimens are quantified by quantitative real-time PCR and western blot. The functional role of Gremlin in NSCLC cells was evaluated by interference RNA (siRNA). The effects of Silenced Gremlin on the resistant PC-9/GR cell line were investigated by proliferation and apoptosis analysis compared with control PC-9 cells. Our results found that Gremlin expression levels were higher in NSCLC tissues, and Gremlin was more highly expressed in PC-9/GR cells compared to PC-9 cells. Knocking down of Gremlin in PC-9/GR cells decreased cell proliferation and increased the expression of BMP7 protein. In addition, Gremlin silencing significantly potentiated apoptosis induced by gefitinib in PC-9/GR with Gremlin knockdown compared to PC-9 transfected with control shRNA, suggesting Gremlin contributes to gefitinib resistance in NSCLC. Gremlin might be explored as a candidate of therapeutic target for modulating EGFR-TKI sensitivity in NSCLC.
肺癌是全球癌症相关死亡的主要原因,其中80%为非小细胞肺癌(NSCLC)病例。Gremlin是一种骨形态发生蛋白(BMP)拮抗剂,在各种癌组织中过表达;然而,关于Gremlin在肺癌发生中的作用知之甚少,Gremlin表达是否与EGFR-TKI耐药相关仍不清楚。在本研究中,通过定量实时PCR和蛋白质印迹法对配对的肿瘤和正常肺标本中Gremlin mRNA和蛋白质的表达进行定量。通过干扰RNA(siRNA)评估Gremlin在NSCLC细胞中的功能作用。与对照PC-9细胞相比,通过增殖和凋亡分析研究沉默Gremlin对耐药PC-9/GR细胞系的影响。我们的结果发现,NSCLC组织中Gremlin表达水平较高,与PC-9细胞相比,PC-9/GR细胞中Gremlin表达更高。敲低PC-9/GR细胞中的Gremlin可降低细胞增殖并增加BMP7蛋白的表达。此外,与转染对照shRNA的PC-9相比,沉默Gremlin显著增强了吉非替尼在敲低Gremlin的PC-9/GR细胞中诱导的凋亡,表明Gremlin促成了NSCLC对吉非替尼的耐药。Gremlin可能是调节NSCLC中EGFR-TKI敏感性的治疗靶点候选物。