Gao Peng, Wang Shijie, Jing Fuchun, Zhan Jiang, Wang Yunhui
Department of Emergency Surgery, The Affiliated Hospital of Qingdao University, Qingdao, Shandong, China.
Department of Gastroenterology, People's Hospital of Juxian, Rizhao, Shandong, China.
Cancer Biomark. 2017;19(1):11-20. doi: 10.3233/CBM-160167.
Growing evidence suggests that microRNA plays an essential role in the development and metastasis of many tumors, including gastric cancer (GC). Expression of microRNA-203 (miR-203) has been reported to decrease in GC. In addition, overexpression of miR-203 inhibits grow of GC cells in vitro and in vivo. However, whether miR-203 affects cell migration and invasion of GC remains unclear. This study aimed to reveal the role of miR-203 on migration and invasion of GC, and its potential mechanisms.
Synthetic pre-miR-203 (miR-203), anti-miR-203 and scrambled negative control RNAs was transfected into the gastric cancer SGC7901 cells to generate miR-203 or anti-miR-203-transfected stable clones. The roles of miR-203 on cell invasion and motility were then analyzed by Transwell migration assay and Wound healing assay in vitro. Using siRNA to targeting ERK1/2, Slug, and E-cadherin or Slug cDNA transfection (to increase Slug expression) to examine the miR-203 signaling pathway. We also examined the efficacies of miR-203 or anti-miR-203 on peritoneal metastasis of SGC7901 cells in the nude mouse model.
Overexpression of miR-203 inhibits SGC7901 cell invasion and motility, followed by decreased phospho-ERK1/2 (pERK1/2) and Slug expression, as well as increased E-cadherin expression. Re-expression of Slug in miR-203/SGC7901cells decreased E-cadherin expression and restored the invasive phenotypes. Targeting E-cadherin in miR-203/SGC7901cells also restored the invasive phenotypes. Inhibition of miR-203 promotes SGC7901 cell invasion and motility, followed by increased phospho-ERK1/2 (pERK1/2) and Slug expression, as well as decreased E-cadherin expression. Targeting ERK1/2 or Slug in anti-miR-203/SGC7901cells increased E-cadherin expression and reversed the invasive phenotypes. In addition, targeting ERK1/2 decreased Slug and increased the E-cadherin expression. Significantly, we found that miR-203 could exert marked inhibition of the peritoneal metastasis of SGC7901 in nude mice in vivo. Targeting miR-203 could exert marked promotion of the peritoneal metastasis of SGC7901 in nude mice in vivo.
miR-203/ERK1/2/Slug/E-cadherin signaling pathway plays an essential role on SGC7901 cell invasion and motility. miR-203 can be novel modalities to prevent peritoneal metastasis of invasive cancers such as gastric cancer.
越来越多的证据表明，微小RNA在包括胃癌（GC）在内的许多肿瘤的发生和转移中起着至关重要的作用。据报道，微小RNA-203（miR-203）在胃癌中的表达降低。此外，miR-203的过表达在体外和体内均抑制胃癌细胞的生长。然而，miR-203是否影响胃癌细胞的迁移和侵袭仍不清楚。本研究旨在揭示miR-203在胃癌迁移和侵袭中的作用及其潜在机制。
将合成的前体miR-203（miR-203）、抗miR-203和乱序阴性对照RNA转染至胃癌SGC7901细胞中，以生成miR-203或抗miR-203转染的稳定克隆。然后通过Transwell迁移试验和体外伤口愈合试验分析miR-203对细胞侵袭和运动的作用。使用靶向ERK1/2、Slug和E-钙黏蛋白的小干扰RNA（siRNA）或转染Slug cDNA（以增加Slug表达）来研究miR-203信号通路。我们还在裸鼠模型中检测了miR-203或抗miR-203对SGC7901细胞腹膜转移的影响。
miR-203的过表达抑制SGC7901细胞的侵袭和运动，随后磷酸化ERK1/2（pERK1/2）和Slug表达降低，以及E-钙黏蛋白表达增加。在miR-203/SGC7901细胞中重新表达Slug可降低E-钙黏蛋白表达并恢复侵袭表型。在miR-203/SGC7901细胞中靶向E-钙黏蛋白也可恢复侵袭表型。抑制miR-203可促进SGC7901细胞的侵袭和运动，随后磷酸化ERK1/2（pERK1/2）和Slug表达增加，以及E-钙黏蛋白表达降低。在抗miR-203/SGC7901细胞中靶向ERK1/2或Slug可增加E-钙黏蛋白表达并逆转侵袭表型。此外，靶向ERK1/2可降低Slug并增加E-钙黏蛋白表达。重要的是，我们发现miR-203在体内可显著抑制裸鼠中SGC7901的腹膜转移。靶向miR-2可在体内显著促进裸鼠中SGC7901的腹膜转移。
miR-203/ERK1/2/Slug/E-钙黏蛋白信号通路在SGC7901细胞的侵袭和运动中起重要作用。miR-203可能是预防胃癌等侵袭性癌症腹膜转移的新方法。
