Gu Lei, Zhang Jiaqiang, Shi Minmin, Peng Chenghong
Department of General Surgery, Ruijin Hospital, School of Medicine, Shanghai Jiao Tong UniversityShanghai 200025, China.
Research Institute of Digestive Surgery, Ruijin Hospital, School of Medicine, Shanghai Jiao Tong UniversityShanghai 200025, China.
Am J Transl Res. 2017 Jun 15;9(6):2798-2806. eCollection 2017.
The aim of this study was to evaluate the effects of miR-1180 on pancreatic cancer. We sampled adjacent cancer and carcinoma tissues from 30 pancreatic cancer patients and measured miR-1180 expression by qRT-PCR and NF-κB protein expression by immunohistochemistry. To explore the effects of this miRNA in cell culture, we used pancreatic cancer (PANC-1) cells that received only vehicle (negative control, NC), miR-1180 mimic, or miR-1180-inhibitor. Cells were treated with cisplatin to induce apoptosis. Proliferation, apoptosis, cell cycle progression, cell invasion, and cell migration were assessed in the three cell groups. The expression levels of relevant proteins (TNIP2, NF-κB, MMP-2, MMP-9, Bax, Bcl-2, p21, and cyclin D1) in each cell group were determined by western blotting. Compared with healthy tissue adjacent to carcinoma tissues, miR-1180 expression in cancer tissues was significantly enhanced (P<0.05). NF-κB protein had a similar expression pattern to miR-1180; miR-1180 expression was positively correlated with NF-κB expression. The invasion and wound healing abilities of miR-1180-inhibited cells were significantly reduced compared with the NC or miR-1180-expressing cells (P<0.05). The cell proliferation rate of miR-1180-inihibited cells was also significantly lower than that of NC or miR-1180-expressing cells (P<0.05), while the cell apoptosis and G1 phase rates of miR-1180-inihibited cells were significantly higher than the NC or miR-1180-expressing cells (P<0.05). In conclusion, suppressing miR-1180 expression may exert anti-cancer effects on pancreatic cancer cells via regulation of TNIP 2/NF-κB signaling and the downstream MMP-2/-9, Bax, Bcl-2, p21, and cyclin D1 factors.
本研究旨在评估miR-1180对胰腺癌的影响。我们采集了30例胰腺癌患者的癌旁组织和癌组织样本，通过qRT-PCR检测miR-1180表达，通过免疫组织化学检测NF-κB蛋白表达。为了在细胞培养中探究这种微小RNA的作用，我们使用了仅接受溶剂（阴性对照，NC）、miR-1180模拟物或miR-1180抑制剂的胰腺癌（PANC-1）细胞。用顺铂处理细胞以诱导凋亡。评估了三个细胞组中的细胞增殖、凋亡、细胞周期进程、细胞侵袭和细胞迁移情况。通过蛋白质免疫印迹法测定每个细胞组中相关蛋白（TNIP2、NF-κB、MMP-2、MMP-9、Bax、Bcl-2、p21和细胞周期蛋白D1）的表达水平。与癌组织相邻的健康组织相比，癌组织中miR-1180表达显著增强（P<0.05）。NF-κB蛋白的表达模式与miR-1180相似；miR-1180表达与NF-κB表达呈正相关。与NC或miR-1180表达细胞相比，miR-1180抑制细胞的侵袭和伤口愈合能力显著降低（P<0.05）。miR-1180抑制细胞的细胞增殖率也显著低于NC或miR-1180表达细胞（P<0.05），而miR-1180抑制细胞的细胞凋亡率和G1期比例显著高于NC或miR-1180表达细胞（P<0.05）。总之，抑制miR-1180表达可能通过调节TNIP 2/NF-κB信号通路及下游MMP-2/-9、Bax、Bcl-2、p21和细胞周期蛋白D1因子对胰腺癌细胞发挥抗癌作用。
