Chen Shi, Zhang Jia-Qiang, Chen Jiang-Zhi, Chen Hui-Xing, Qiu Fu-Nan, Yan Mao-Lin, Chen Yan-Ling, Peng Cheng-Hong, Tian Yi-Feng, Wang Yao-Dong
Department of Hepatobiliary Surgery, Fujian Provincial Hospital, Fujian Medical University, Fuzhou 350001, P.R. China; Department of General Surgery, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200025, PR China; Research Institute of Digestive Surgery, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China.
Department of General Surgery, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200025, PR China; Research Institute of Digestive Surgery, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China.
Int J Biol Macromol. 2017 Sep;102:718-728. doi: 10.1016/j.ijbiomac.2017.03.123. Epub 2017 Mar 23.
This study aims to investigate the roles of lncRNA ANRIL in epithelial-mesenchymal transition (EMT) by regulating the ATM-E2F1 signaling pathway in pancreatic cancer (PC). PC rat models were established and ANRIL overexpression and interference plasmids were transfected. The expression of ANRIL, EMT markers (E-cadherin, N-cadherin and Vimentin) and ATM-E2F1 signaling pathway-related proteins (ATM, E2F1, INK4A, INK4B and ARF) were detected. Small molecule drugs were applied to activate and inhibit the ATM-E2F1 signaling pathway. Transwell assay and the scratch test were adopted to detect cell invasion and migration abilities. ANRIL expression in the PC cells was higher than in normal pancreatic duct epithelial cells. In the PC rat models and PC cells, ANRIL interference promoted the expressions of INK4B, INK4A, ARF and E-cadherin, while reduced N-cadherin and Vimentin expression. Over-expressed ANRIL decreased the expression of INK4B, INK4A, ARF and E-cadherin, but raised N-cadherin and Vimentin expressions. By inhibiting the ATM-E2F1 signaling pathway in PC cells, E-cadherin expression increased but N-cadherin and Vimentin expressions decreased. After ANRIL was silenced or the ATM-E2F1 signaling pathway inhibited, PC cell migration and invasion abilities were decreased. In conclusion, over-expression of lncRNA ANRIL can promote EMT of PC cells by activating the ATM-E2F1 signaling pathway.
本研究旨在通过调控胰腺癌（PC）中的ATM-E2F1信号通路来探讨长链非编码RNA ANRIL在上皮-间质转化（EMT）中的作用。建立PC大鼠模型并转染ANRIL过表达和干扰质粒。检测ANRIL、EMT标志物（E-钙黏蛋白、N-钙黏蛋白和波形蛋白）以及ATM-E2F1信号通路相关蛋白（ATM、E2F1、INK4A、INK4B和ARF）的表达。应用小分子药物激活和抑制ATM-E2F1信号通路。采用Transwell实验和划痕实验检测细胞侵袭和迁移能力。PC细胞中ANRIL的表达高于正常胰腺导管上皮细胞。在PC大鼠模型和PC细胞中，ANRIL干扰促进了INK4B、INK4A、ARF和E-钙黏蛋白的表达，同时降低了N-钙黏蛋白和波形蛋白的表达。过表达ANRIL降低了INK4B、INK4A、ARF和E-钙黏蛋白的表达，但提高了N-钙黏蛋白和波形蛋白的表达。通过抑制PC细胞中的ATM-E2F1信号通路，E-钙黏蛋白表达增加，但N-钙黏蛋白和波形蛋白表达降低。在ANRIL沉默或ATM-E2F1信号通路被抑制后，PC细胞的迁移和侵袭能力降低。总之，lncRNA ANRIL的过表达可通过激活ATM-E2F1信号通路促进PC细胞的EMT。
