Department of General Surgery, Sir Run Run Shaw Hospital, School of Medicine, Zhejiang University, Hangzhou, China.
Medical Technology College, Zhejiang Chinese Medical University, Hangzhou, China.
Acta Biochim Biophys Sin (Shanghai). 2019 Aug 5;51(8):814-825. doi: 10.1093/abbs/gmz071.
Recent studies have demonstrated that the expression of the long non-coding RNA (lncRNA) AFAP1-AS1 in pancreatic cancer is negatively correlated with survival and prognosis. However, the effects of oridonin and lncRNA AFAP1-AS1 on the epithelial-to-mesenchymal transition (EMT) and migration of pancreatic cancer cells have not been fully elucidated. Surgery is the only potentially curative method for pancreatic cancer, but postoperative recurrence and metastasis are common. The aim of the present study was to assess the effect of oridonin and lncRNA AFAP1-AS1 silencing on pancreatic cancer cells. The pancreatic cancer cell lines BxPC-3 and PANC-1 cells were transfected with siAFAP1-AS1 and its negative control (siNC). After that, oridonin was used to treat the siAFAP1-AS1-transfected cells. The expression of lncRNA AFAP1-AS1 was downregulated in the pancreatic cancer cell lines BxPC-3 and PANC-1. The apoptosis and cell cycle progression of pancreatic cancer cells were evaluated by flow cytometry and Hoechst 33258 staining. Metastasis and invasion of BxPC-3 and PANC-1 cells were detected by transwell migration assay, real-time cell analysis, and western blot analysis. Cells were transfected with the lentiviral siAFAP1-AS1 and siNC, and tumorigenesis was evaluated in BALB/C nude mice. Immunohistochemical examination was used to verify the effects of oridonin and siAFAP1-AS1 on pancreatic cancer. The results demonstrated that the combination of oridonin and siAFAP1-AS1 inhibited pancreatic cancer cell proliferation, induced apoptosis, arrested cell cycle progression, prevented the migration, regulated EMT-related protein expression in BxPC-3 and PANC-1 cells, and inhibited pancreatic cancer cell tumorigenicity and EMT in nude mice.
最近的研究表明，长链非编码 RNA (lncRNA) AFAP1-AS1 在胰腺癌中的表达与生存和预后呈负相关。然而，冬凌草甲素和 lncRNA AFAP1-AS1 对胰腺癌细胞上皮-间质转化 (EMT) 和迁移的影响尚未完全阐明。手术是胰腺癌唯一有潜在治愈可能的方法，但术后复发和转移较为常见。本研究旨在评估冬凌草甲素和 lncRNA AFAP1-AS1 沉默对胰腺癌细胞的影响。用 siAFAP1-AS1 和其阴性对照 (siNC) 转染胰腺癌细胞系 BxPC-3 和 PANC-1 细胞。之后，用冬凌草甲素处理转染 siAFAP1-AS1 的细胞。下调胰腺癌细胞系 BxPC-3 和 PANC-1 中 lncRNA AFAP1-AS1 的表达。通过流式细胞术和 Hoechst 33258 染色评估胰腺癌细胞的凋亡和细胞周期进程。通过 Transwell 迁移实验、实时细胞分析和 Western blot 分析检测 BxPC-3 和 PANC-1 细胞的转移和侵袭。用慢病毒 siAFAP1-AS1 和 siNC 转染细胞，并在 BALB/C 裸鼠中评估肿瘤生成。免疫组织化学检测用于验证冬凌草甲素和 siAFAP1-AS1 对胰腺癌的影响。结果表明，冬凌草甲素和 siAFAP1-AS1 的联合抑制了胰腺癌细胞的增殖，诱导了细胞凋亡，阻滞了细胞周期进程，阻止了 BxPC-3 和 PANC-1 细胞的迁移，调节了 EMT 相关蛋白的表达，并抑制了裸鼠胰腺癌细胞的肿瘤生成和 EMT。
