1. 郑州大学第一附属医院, a. 麻醉科; b. 消化内科, 郑州 450052; 2. 河南省疾病预防控制中心门诊部, 郑州 450003; 3. 郑州大学第五附属医院麻醉科, 郑州 450052
Etomidate Inhibits Proliferation, Migration and Invasion of Gastric Cancer Cells by Regulating the Expression of microRNA-211-5p/ROBO1
WANG Dong-mei1a, XU Bi-yu2, YANG Tao3, HE Juan1a, ZHAO Hua-ping1a, LÜ Shuai1b, XING Na1a*
1a. Department of Anesthesiology; 1b. Department of Gastroenterology, First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China; 2. Outpatient Department of Henan Disease Control Center, Zhengzhou 450003, China; 3. Department of Anesthesiology, Fifth Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China
Abstract：OBJECTIVE To explore the mechanism of etomidate for inhibiting the proliferation, migration and invasion of gastric cancer cells by regulating the expression of miR-211-5p/ROBO1. METHODS QRT-PCR and Western blot were used to detect the effects of etomidate on the expression of microRNA-211-5p and ROBO1 in gastric cancer AGS and MNK-45 cells. MTT assay was used to detect the effect of etomidate on cell proliferation. Transwell assay was used to detect the effect of etomidate on cell invasion and migration. Double luciferase reporter assay and Western blot were used to detect the relationship between microRNA-211-5p and ROBO1. The effects of over-expression of microRNA-211-5p or inhibition of ROBO1 expression on proliferation, invasion and migration of gastric cancer AGS and MNK-45 cells were observed. RESULTS Compared with the control group, the inhibition rate of proliferation and the levels of microRNA-211-5p and p21 of gastric cancer AGS and MNK-45 cells in etomidate 0.2, 0.4 and 0.8 μg·mL-1 groups increased in a dose-dependent manner, while the number of migrating and invading cells, ROBO1, cyclin D1, MMP-2 and MMP-9 levels decreased in a dose-dependent manner (P<0.05). TargetScan bioinformatics software predicted that there were complementary sequences between microRNA-211-5p and ROBO1 3′UTR. Double luciferase assay showed that there were specific binding sites between microRNA-211-5p and ROBO1. Western blot analysis showed that microRNA-211-5p negatively regulated ROBO1 expression. Compared with microRNA-NC, the proliferation inhibition rate and p21 level of gastric cancer AGS and MNK-45 cells in microRNA-211-5p group increased significantly, while the number of migration and invasion cells, cyclin D1, MMP-2 and MMP-9 protein levels decreased significantly (P<0.05). Compared with si-NC, after inhibiting ROBO1 expression, the proliferation inhibition rate and p21 level of AGS and MNK-45 cells in si-ROBO1 group increased significantly, while the number of migrating and invading cells, ROBO1, cyclin D1, MMP-2 and MMP-9 protein levels decreased significantly (P<0.05). Compared with etomidate + anti-microRNA-NC group, the inhibitory rates of microRNA-211-5p, p21 and cell proliferation in gastric cancer AGS and MNK-45 cells in etomidate + anti-microRNA-211-5p group decreased significantly, while the number of migrating and invading cells, ROBO1, cyclin D1, MMP-2 and MMP-9 protein levels increased significantly (P<0.05). CONCLUSION Etomidate can inhibit the proliferation, invasion and migration of gastric cancer cells, which may play a role in inhibiting the proliferation, migration and invasion of gastric cancer cells by regulating the expression of microRNA-211-5p/ROBO1.
王冬梅, 徐碧钰, 杨涛, 何娟, 赵华平, 吕帅, 邢娜. 依托咪酯通过调控miR-211-5p/ROBO1的表达抑制胃癌细胞增殖、迁移和侵袭的机制研究[J]. 中国药学杂志, 2020, 55(20): 1686-1695.
WANG Dong-mei, XU Bi-yu, YANG Tao, HE Juan, ZHAO Hua-ping, LÜ Shuai, XING Na. Etomidate Inhibits Proliferation, Migration and Invasion of Gastric Cancer Cells by Regulating the Expression of microRNA-211-5p/ROBO1. Chinese Pharmaceutical Journal, 2020, 55(20): 1686-1695.
LIU W W, WANG F, ZHANG Y, et al. Progress in the study of the mechanism of competitive endogenous RNA and gastric cancer. Chin J Lab Med (中华检验医学杂志), 2017, 40(8):637-640.
[2]
WANG Y Y. Study on apoptosis of human hepatoma HepG2 cells induced by different concentrations of etomidate. Taiyuan: Shanxi Medical University, 2014.
[3]
WANG Q X, ZHU Y Q, ZHANG H, et al. Altered MiRNA expression in gastric cancer: a systematic review and Meta-analysis. Cell Physiol Biochem, 2015, 35(3):933-944.
[4]
WANG L, SHEN Y F, SHI Z M, et al. Overexpression miR-211-5p hinders the proliferation, migration, and invasion of thyroid tumor cells by downregulating SOX11. J Clin Lab Anal, 2017, 32(3):e22293.
[5]
WANG K, JIN W, JIN P, et al. miR-211-5p suppresses Metastatic behavior by targeting SNAI1 in renal cancer. Mol Cancer Res, 2017, 15(4):448-456.
[6]
ZHANG C X, MENG J, XU D P, et al. Expression and clinical significance of Robo 1 protein in gastric cancer. J Cont Med (当代医学), 2016, 22(17):3-5.
[7]
SALI, M TAHIR A A, KHAN, et al. Etomidate suppresses invasion and migration of human A549 lung adenocarcinoma cells. Anticancer Res, 2019, 39(1):215-223.
[8]
CAO Q, LIU F, JI K, et al. MicroRNA-381 inhibits the metastasis of gastric cancer by targeting TMEM16A expression. J Exp Clin Cancer Res, 2017, 36(1):29-44.
[9]
LIU W X. Effects of different doses of dexmedetomidine combined with etomidate on pain and infection in patients after radical gastrectomy . J Pract Cancer (实用癌症杂志), 2016, 31(12):2023-2025.
[10]
JIAO Y X. The apoptosis of HL-60 cells induced by etomidate and the inhibitory effect of pretreatment on it . Dalian: Dalian Medical University, 2007.
[11]
ZHENG L, JIAO W, MEI H, et al. miRNA-337-3p inhibits gastric cancer progression through repressing myeloid zinc finger 1-facilitated expression of matrix metalloproteinase 14. Oncotarget, 2016, 7(26):40314-40328.
[12]
YANG L, LIANG H, WANG Y, et al. MiRNA-203 suppresses tumor cell proliferation, migration and invasion by targeting Slug in gastric cancer. Protein Cell, 2016, 7(5):383-387.
[13]
DONG L, DENG J, SUN Z M, et al. Interference with the β-catenin gene in gastric cancer induces changes to the miRNA expression profile. Tumour Biol, 2015, 36(9):6973-6983.
[14]
DÍAZMARTÍNEZ M, BENITOJARDÓN L, ALONSO L, et al. miR-204-5p and miR-211-5p contribute to BRAF inhibitor resistance in melanoma. Cancer Res, 2018, 78(4):1017-1030.
[15]
QUAN J, PAN X, HE T, et al. Tumor suppressor miR-211-5p is associated with cellular migration, proliferation and apoptosis in renal cell carcinoma. Exp Ther Med, 2018, 15(4):4019-4028.
[16]
CHEN L, ZHANG Z, YI Z, et al. MicroRNA-211-5p suppresses tumour cell proliferation, invasion, migration and metastasis in triple-negative breast cancer by directly targeting SETBP1. Br J Cancer, 2017, 117(1):78-88.
[17]
QIN F, ZHANG H, MA L, et al. Low expression of Slit2 and robo1 is associated with poor prognosis and brain-specific metastasis of breast cancer patients. Sci Rep, 2015, 5(1):14430-14441.
[18]
WANG J, WANG L, LIU F F, et al. Expression of Robo1 protein in breast cancer and its relationship with brain metastasis of breast cancer. Chin J Oncol (中华肿瘤杂志), 2011, 33(6):447-451.
[19]
LIU Z G, LIU F, HAN J H. Effect of siRNA targeting interference of ROBO1 gene expression on cell cycle, proliferation and related proteins in esophageal cancer cells. Chin J Geront (中国老年学杂志), 2018, 38(17):4216-4219.
[20]
CHEN W, YE L, WEN D, et al. MiR-490-5p inhibits hepatocellular carcinoma cell proliferation, migration and invasion by directly regulating ROBO1. Pathol Oncol Res, 2017, 25(1):1-9.
2020, Vol. 55 
