Zhang Nianliang, Xing Xiangji, Gu Fengcai, Zhou Gang, Liu Xianglan, Li Baoqiang
Department of Anesthesiology, Rizhao People's Hospital, Rizhao 276826, Shandong, People's Republic of China.
Department of Anesthesiology, Rizhao Women and Children Hospital, Rizhao 276800, Shandong, People's Republic of China.
Onco Targets Ther. 2020 Jun 9;13:5309-5321. doi: 10.2147/OTT.S244550. eCollection 2020.
Metastasis remains one of the greatest challenges involved in treating gastric cancer (GC). Ropivacaine (Rop) is not only a well-documented local anesthetic medicament but also has been reported to exert an antitumor role in cancer development. This study explored the effects of ropivacaine on the growth, migration and invasion of gastric cancer and the underlying mechanisms.
Cell Counting Kit-8 (CCK8) assay was conducted to test the effect of Rop on the proliferation of AGS and BGC-823 GC cells. Moreover, cell apoptosis, migration and invasion were examined by flow cytometry and transwell assay, respectively. The expression of miR-520a-3p was determined by qRT-PCR. miRNA targeting sites were analyzed using bioinformatics analysis and dual-luciferase reporter assay. Protein levels of WEE1 and PI3K/AKT were detected by Western blot. Furthermore, the tumor-forming experiment of nude mice was used to detect the growth of cells in vivo.
Rop inhibited proliferation but promoted apoptosis of GC cells. Besides, the migration and invasion of GC cells were also inhibited by Rop. Moreover, miR-520a-3p expression was enhanced by Rop, and transfection with miR-520a-3p mimic decreased cell proliferation, migration and invasion. The upregulation of miR-520a-3p was partly contributed to the inhibitory effect of ropivacaine on GC cell lines. Finally, Rop inactivated WEE1 and PI3K/AKT pathway via upregulation of miR-520a-3p.
Our results suggested that Rop decreased growth, migration and invasion of GC cells via regulating miR-520a-3p expression and further inactivated WEE1 and PI3K/AKT signaling pathways.
转移仍然是胃癌（GC）治疗中面临的最大挑战之一。罗哌卡因（Rop）不仅是一种有充分文献记载的局部麻醉药物，而且据报道在癌症发展过程中具有抗肿瘤作用。本研究探讨了罗哌卡因对胃癌生长、迁移和侵袭的影响及其潜在机制。
采用细胞计数试剂盒-8（CCK8）检测罗哌卡因对AGS和BGC-823胃癌细胞增殖的影响。此外，分别通过流式细胞术和Transwell实验检测细胞凋亡、迁移和侵袭情况。采用qRT-PCR检测miR-520a-3p的表达。通过生物信息学分析和双荧光素酶报告基因检测分析miRNA靶向位点。采用蛋白质免疫印迹法检测WEE1和PI3K/AKT的蛋白水平。此外，利用裸鼠成瘤实验检测细胞在体内的生长情况。
罗哌卡因抑制胃癌细胞增殖，但促进其凋亡。此外，罗哌卡因还抑制胃癌细胞的迁移和侵袭。而且，罗哌卡因可增强miR-520a-3p的表达，转染miR-520a-3p模拟物可降低细胞增殖、迁移和侵袭能力。miR-520a-3p的上调部分归因于罗哌卡因对胃癌细胞系的抑制作用。最后，罗哌卡因通过上调miR-520a-3p使WEE1和PI3K/AKT通路失活。
我们的结果表明，罗哌卡因通过调节miR-520a-3p的表达降低胃癌细胞的生长、迁移和侵袭能力，并进一步使WEE1和PI3K/AKT信号通路失活。
