Nie Meng-Lin, Han Jing, Huang Han-Chen, Guo Ting, Huangfu Long-Tao, Cheng Xiao-Jing, Li Xiao-Mei, Du Hong, Li Qing-Da, Wen Xian-Zi, Ji Jia-Fu
Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education), Division of Gastrointestinal Cancer Translational Research Laboratory, Peking University Cancer Hospital & Institute, Beijing, People's Republic of China.
Key Laboratory of RNA Biology, Beijing Key Laboratory of Noncoding RNA, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, People's Republic of China.
Cancer Manag Res. 2019 Jun 12;11:5375-5391. doi: 10.2147/CMAR.S201793. eCollection 2019.
Emerging evidence has shown that long noncoding RNAs (lncRNAs) participate in oncogenesis and tumor progression. We previously found a novel lncRNA p4516 which was closely associated with prognosis by preliminary study of lncRNA expression profile from paired tumors and nontumor tissues in 198 gastric cancer (GC) patients. However, the exact biological functions and the underlying molecular mechanisms of p4516 in gastric tumorigenesis still remain unclear. The RNA fluorescence in situ hybridization (RNA-FISH) analysis, cytoplasmic and nuclear RNA isolation and qRT-PCR were applied to determine the subcellular localization of p4516. Expression levels of p4516 were assessed using qRT-PCR in both GC cell lines and in 142 primary GC tissues. Correlations between p4516 expression and GC patients' clinicopathological parameters were analyzed. Gain- and loss-of-function experiments were employed to investigate the role of p4516 in proliferation, migration and invasion both in vitro and in vivo. In addition, Western blotting and immunohistochemical staining were used to examine the protein expression levels. LncRNA p4516 was mainly localized in the nucleus of GC cells and p4516 tended to have higher expression levels in GC cells compared to the normal gastric mucosa-derived cells GES-1. Furthermore, higher expression levels of p4516 correlated with worse clinical outcomes in GC patients and acted as an independent prognostic biomarker. Functional analysis revealed that p4516 participated in the regulation of GC cell proliferation, invasion and migration both in vivo and in vitro. Moreover, p4516 was involved in epithelial-mesenchymal transition (EMT) in GC cells. Our study demonstrated the oncogenic role of novel lncRNA p4516 in the gastric carcinogenesis for the first time. High expression of p4516 may act as prognostic marker in patient with gastric cancer.
新出现的证据表明，长链非编码RNA（lncRNA）参与肿瘤发生和肿瘤进展。我们之前通过对198例胃癌（GC）患者配对肿瘤组织和非肿瘤组织的lncRNA表达谱进行初步研究，发现了一种与预后密切相关的新型lncRNA p4516。然而，p4516在胃癌发生中的具体生物学功能和潜在分子机制仍不清楚。应用RNA荧光原位杂交（RNA-FISH）分析、细胞质和细胞核RNA分离以及qRT-PCR来确定p4516的亚细胞定位。使用qRT-PCR评估p4516在GC细胞系和142例原发性GC组织中的表达水平。分析p4516表达与GC患者临床病理参数之间的相关性。采用功能获得和功能缺失实验来研究p4516在体外和体内对增殖、迁移和侵袭的作用。此外，使用蛋白质印迹法和免疫组织化学染色来检测蛋白质表达水平。lncRNA p4516主要定位于GC细胞的细胞核中，与正常胃黏膜来源的细胞GES-1相比，p4516在GC细胞中的表达水平往往更高。此外，p4516的高表达水平与GC患者较差的临床结局相关，并作为独立的预后生物标志物。功能分析表明，p4516在体内和体外均参与GC细胞增殖、侵袭和迁移的调控。此外，p4516参与了GC细胞的上皮-间质转化（EMT）。我们的研究首次证明了新型lncRNA p4516在胃癌发生中的致癌作用。p4516的高表达可能作为胃癌患者的预后标志物。
