Zang Wenqiao, Wang Yuanyuan, Wang Tao, Du Yuwen, Chen Xiaonan, Li Min, Zhao Guoqiang
College of Basic Medical Sciences, Zhengzhou University, Zhengzhou, 450001, Henan Province, China.
Department of Hemato-tumor, The First Affiliated Hospital of Henan University of TCM, Zhengzhou, 450000, China.
Mol Cancer. 2015 Feb 13;14:37. doi: 10.1186/s12943-015-0315-3.
miR-663 is associated with many important biologic processes, such as the evolution, development, viral infection, inflammatory response, and carcinogenesis among vertebrates. However, the molecular function and mechanism of miR-663 in pancreatic cancer growth and invasion is still unclear.
Western blot and real-time PCR were used to study the expression level of eEF1A2 protein and miR-663 in pancreatic cancer tissues and cell lines. The Pearson χ (2) test was used to determine the correlation between miR-663 expression and clinicopathologic features of patients. Patients' survival was analyzed using the Kaplan-Meier method, using the log-rank test for comparison. The biological function of miR-663 was examined by measuring cell growth, cell invasion and apoptosis analysis in vitro and in vivo. miR-663 target gene and signaling pathway was identified by luciferase activity assay and western blot.
We found that, in pancreatic cancer, eEF1A2 was significantly upregulated but miR-663 was significantly downregulated. Further results showed that the expression level of eEF1A2 and miR-663 was strongly associated with TNM stage and node metastasis status of the patients. miR-663 and eEF1A2 were inversely correlated with each other, and the changes in the expression levels of each can also predict the survival of patients with pancreatic cancer. We identified miR-663 as a tumor attenuate molecular that attenuated the proliferation and invasion of pancreatic cancer cells both in vitro and in vivo. Finally, we confirmed that the expression of eEF1A2 can partially restore the pro-apoptotic and anti-invasion functions of miR-663.
miR-663 attenuated the proliferation and invasion of pancreatic cells both in vitro and in vivo by directly targeting eEF1A2. miR-663 and eEF1A2 might be potential targets for the treatment of pancreatic cancer in the future.
miR-663与许多重要的生物学过程相关，如脊椎动物的进化、发育、病毒感染、炎症反应和致癌作用。然而，miR-663在胰腺癌生长和侵袭中的分子功能及机制仍不清楚。
采用蛋白质免疫印迹法和实时定量PCR研究胰腺癌组织及细胞系中eEF1A2蛋白和miR-663的表达水平。采用Pearson χ²检验确定miR-663表达与患者临床病理特征之间的相关性。采用Kaplan-Meier法分析患者生存率，用对数秩检验进行比较。通过体外和体内测量细胞生长、细胞侵袭及凋亡分析来检测miR-663的生物学功能。通过荧光素酶活性测定和蛋白质免疫印迹法鉴定miR-663靶基因及信号通路。
我们发现，在胰腺癌中，eEF1A2显著上调而miR-663显著下调。进一步结果表明，eEF1A2和miR-663的表达水平与患者的TNM分期及淋巴结转移状态密切相关。miR-663与eEF1A2呈负相关，且二者表达水平的变化均可预测胰腺癌患者的生存率。我们确定miR-663为一种肿瘤抑制分子，其在体外和体内均可减弱胰腺癌细胞的增殖和侵袭。最后，我们证实eEF1A2的表达可部分恢复miR-663的促凋亡和抗侵袭功能。
miR-663通过直接靶向eEF1A2在体外和体内减弱胰腺细胞的增殖和侵袭。miR-663和eEF1A2可能是未来胰腺癌治疗的潜在靶点。
