Zhao Huazhou, Zhang Kecheng, Wang Ting, Cui Jianxin, Xi Hongqing, Wang Yi, Song Yanjing, Zhao Xudong, Wei Bo, Chen Lin
Department of General Surgery, General Hospital of Chinese People's Liberation Army, Beijing 100853, P.R. China.
Department of General Surgery, 309th Hospital of Chinese People's Liberation Army, Beijing 100091, P.R. China.
Oncol Lett. 2018 Jun;15(6):8620-8626. doi: 10.3892/ol.2018.8389. Epub 2018 Mar 30.
Gastric cancer (GC) is the second-leading cause of cancer-associated mortality worldwide. AFAP1-antisense RNA 1 (AFAP1-AS1), a long non-coding RNA (lncRNA), is believed to promote the aggressive progression of cancer; however, its role in GC remains largely unknown. In the present study, the expression of AFAP1-AS1 in GC tissues and cell lines was measured using reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Knockdown of AFAP1-AS1 was performed using a lentiviral vector containing a short hairpin RNA. The proliferation of GC cells was measured using Cell Counting kit-8. The migration and invasion of GC cells were analyzed using a QCM Laminin Migration Assay kit and a Cell Invasion Assay kit. The levels of epithelial-mesenchymal transition (EMT)-associated proteins were detected by western blot analysis. The cut-off value of the expression of AFAP1-AS1 was evaluated using receiver operating characteristic (ROC) curves and patient survival rate was analyzed using Kaplan-Meier. The expression of AFAP1-AS1 was significantly increased in the primary tumor tissues of GC patients with lymph node metastasis or tumor node metastasis stage (stage III or IV; P<0.01). ROC curve analysis revealed that the expression of AFAP-AS1, at a cut-off value of 0.5040, could distinguish GC tissues from the matched normal tissues, with an AUC of 0.8802, sensitivity of 81.25% and specificity of 83.75%. The overexpression of AFAP1-AS1 was positively associated with the poor survival rates of GC patients. Furthermore, the downregulation of AFAP1-AS1 significantly inhibited the proliferation, migration and invasion of GC cells (P<0.01). The decrease in AFAP1-AS1 expression significantly suppressed the expression level of N-cadherin protein in GC cells and increased that of E-cadherin. The present study demonstrated that the expression signature of AFAP1-AS1 may serve as a biomarker for the diagnosis and prognosis of GC, and its downregulation may repress the aggressive progression of GC, partially through inhibiting the EMT progress.
胃癌（GC）是全球癌症相关死亡的第二大原因。AFAP1反义RNA 1（AFAP1-AS1）是一种长链非编码RNA（lncRNA），被认为可促进癌症的侵袭性进展；然而，其在胃癌中的作用仍 largely未知。在本研究中，使用逆转录定量聚合酶链反应（RT-qPCR）检测AFAP1-AS1在胃癌组织和细胞系中的表达。使用含有短发夹RNA的慢病毒载体进行AFAP1-AS1的敲低。使用细胞计数试剂盒-8检测胃癌细胞的增殖。使用QCM层粘连蛋白迁移分析试剂盒和细胞侵袭分析试剂盒分析胃癌细胞的迁移和侵袭。通过蛋白质印迹分析检测上皮-间质转化（EMT）相关蛋白的水平。使用受试者工作特征（ROC）曲线评估AFAP1-AS1表达的临界值，并使用Kaplan-Meier分析患者生存率。AFAP1-AS1的表达在伴有淋巴结转移或肿瘤淋巴结转移分期（III期或IV期；P<0.01）的胃癌患者的原发性肿瘤组织中显著增加。ROC曲线分析显示，AFAP-AS1的表达在临界值为0.5040时，可区分胃癌组织与匹配的正常组织，曲线下面积（AUC）为0.8802，灵敏度为81.25%，特异性为83.75%。AFAP1-AS1的过表达与胃癌患者的低生存率呈正相关。此外，AFAP1-AS1的下调显著抑制了胃癌细胞的增殖、迁移和侵袭（P<0.01）。AFAP1-AS1表达的降低显著抑制了胃癌细胞中N-钙黏蛋白的表达水平，并增加了E-钙黏蛋白的表达。本研究表明，AFAP1-AS1的表达特征可能作为胃癌诊断和预后的生物标志物，其下调可能部分通过抑制EMT进程来抑制胃癌的侵袭性进展。
