Zhu Ping, Ge Nan, Liu Dongyan, Yang Fan, Zhang Kai, Guo Jintao, Liu Xiang, Wang Sheng, Wang Guoxin, Sun Siyu
Endoscopy Center, Shengjing Hospital of China Medical University, Shenyang, Liaoning 110004, P.R. China.
Oncol Lett. 2018 Jul;16(1):603-611. doi: 10.3892/ol.2018.8652. Epub 2018 May 7.
The incidence of pancreatic cancer is increasing annually in Asia as a whole. Pancreatic cancer ranks sixth in terms of incidence of all malignant tumors. Circular RNA (circRNA) is a type of non-coding RNA which forms a covalently closed continuous loop. CircRNA is extensively expressed in the cytoplasm, and is markedly conservative and stable. MicroRNA (miR)-378a-3p and human (hsa)_circ_0006215 were detected using the reverse transcription-quantitative polymerase chain reaction (RT-qPCR) in tissue and cells. Western blot analysis detected the SERPINA4 and hsa_circ_0006215 expression in tissue. A Cell Counting Kit-8 assay was used to determine cell stability. Flow cytometry was used to determine the cell apoptotic rate. Transwell assays were used to determine cell migration. hsa_circ_0006215 was identified as a significantly upregulated circRNA. RT-qPCR results verified that, in 30 samples of pancreatic cancer tissue and paracancerous tissue, hsa_circ_0006215 expression was increased in pancreatic cancer tissue, miR-378a-3p expression was decreased in pancreatic cancer tissue, and SERPINA4 expression was increased in pancreatic cancer tissue (P<0.05). Using bioinformatics database and bioinformatics analysis, the interaction network of hsa_circ_0006215 indicated that this circRNA was most likely to regulate the expression of miR-378a-3p. Further interaction analysis revealed that the SERPINA4 gene was a regulatory target gene most likely to have an influence. The present study identified the effects of hsa_circ_0006215, miR-378a-3p and SERPINA4 signaling pathways in pancreatic cancer cells.
在整个亚洲,胰腺癌的发病率逐年上升。在所有恶性肿瘤中,胰腺癌的发病率排名第六。环状RNA(circRNA)是一种非编码RNA,其形成共价闭合的连续环。circRNA在细胞质中广泛表达,且具有明显的保守性和稳定性。采用逆转录-定量聚合酶链反应(RT-qPCR)检测组织和细胞中的微小RNA(miR)-378a-3p和人(hsa)_circ_0006215。蛋白质免疫印迹分析检测组织中丝氨酸蛋白酶抑制剂A4(SERPINA4)和hsa_circ_0006215的表达。采用细胞计数试剂盒-8法测定细胞稳定性。流式细胞术用于测定细胞凋亡率。Transwell实验用于测定细胞迁移。hsa_circ_0006215被鉴定为显著上调的circRNA。RT-qPCR结果证实,在30例胰腺癌组织和癌旁组织样本中,胰腺癌组织中hsa_circ_0006215表达升高,miR-378a-3p表达降低,SERPINA4表达升高(P<0.05)。通过生物信息学数据库和生物信息学分析,hsa_circ_0006215的相互作用网络表明,该circRNA最有可能调控miR-378a-3p的表达。进一步的相互作用分析显示,SERPINA4基因是最有可能产生影响的调控靶基因。本研究确定了hsa_circ_0006215、miR-378a-3p和SERPINA4信号通路在胰腺癌细胞中的作用。