Pan Yan, Chen Jing, Tao Leilei, Zhang Kai, Wang Rui, Chu Xiaoyuan, Chen Longbang
Department of Medical Oncology, Jinling Hospital, School of Medicine, Nanjing University, Nanjing, Jiangsu, China.
Oncotarget. 2017 May 16;8(20):33144-33158. doi: 10.18632/oncotarget.16562.
Emerging evidence indicates that the dysregulation of long non-coding RNAs (lncRNAs) contributes to the development and progression of lung adenocarcinoma (LAD), however the underlying mechanism of action of lncRNAs remains unclear. It is well known that the effective treatment of cancers has been hindered by drug resistance in the clinical setting. Epithelial-mesenchymal transition (EMT) has been recognized to be involved in acquiring drug resistance, cell migration and invasion properties in several types of cancer. Docetaxel-resistant LAD cells established previously in our lab present chemoresistant and mesenchymal features. Long intergenic non-protein coding RNA, regulator of reprogramming (linc-ROR), was first discovered in induced pluripotent stem cells (iPSCs) and was upregulated in docetaxel-resistant LAD cells. In this study, we tried to make clarification of lincRNA-related mechanisms underlying EMT followed by acquired resistance to chemotherapy in LAD. In order to hit the mark, we made use of multiple methods including microarray analysis, qRT-PCR, western blotting analysis, loss/gain-of-function analysis, luciferase assays, drug sensitivity assays, wound-healing assay and invasion assay. We found that decreased expression of linc-ROR effectively reversed EMT in docetaxel-resistant LAD cells and sensitized them to chemotherapy. The function of linc-ROR exerted in LAD cells depended on the sponging of miR-145, therefore, releasing the miR-145 target FSCN1, and thus contributing to the acquisition of chemoresistance and EMT phenotypes of docetaxel-resistant LAD cells. Our findings revealed that linc-ROR might act as potential therapeutic target to overcome chemotherapy resistance in LAD.
新出现的证据表明，长链非编码RNA（lncRNA）的失调促成了肺腺癌（LAD）的发生和发展，然而lncRNA的潜在作用机制仍不清楚。众所周知，癌症的有效治疗在临床中受到耐药性的阻碍。上皮-间质转化（EMT）已被认为与几种类型癌症中获得耐药性、细胞迁移和侵袭特性有关。我们实验室先前建立的多西他赛耐药LAD细胞具有化学抗性和间质特征。长链基因间非编码RNA，重编程调节因子（linc-ROR），最初在诱导多能干细胞（iPSC）中被发现，并且在多西他赛耐药LAD细胞中上调。在本研究中，我们试图阐明LAD中EMT以及随后获得化疗耐药性背后的lincRNA相关机制。为了达到目的，我们使用了多种方法，包括微阵列分析、qRT-PCR、蛋白质免疫印迹分析、功能缺失/获得分析、荧光素酶测定、药物敏感性测定、伤口愈合测定和侵袭测定。我们发现，linc-ROR表达降低有效地逆转了多西他赛耐药LAD细胞中的EMT，并使它们对化疗敏感。linc-ROR在LAD细胞中发挥的功能依赖于对miR-145的海绵吸附作用，因此，释放miR-145的靶标FSCN1，从而促成多西他赛耐药LAD细胞获得化疗耐药性和EMT表型。我们的研究结果表明，linc-ROR可能作为克服LAD化疗耐药性的潜在治疗靶点。
