作者：赵轶峰1，张超2，胡晓敏1，王雄1，赵铁军3，李曙光1

单位：1.河北北方学院附属第一医院 胃肠肿瘤外科，河北 张家口 075000；2.河北北方学院附属第一医院 介入科，河北 张家口 075000；3.河北北方学院基础医学院，河北 张家口 075000

Authors: Zhao Yifeng1, Zhang Chao2, Hu Xiaomin1, Wang Xiong1, Zhao Tiejun3, Li Shuguang1

Unit: 1.Department of Gastrointestinal Tumor Surgery, the First Hospital Affiliated to Hebei North University, Zhangjiakou 075000, Hebei, China；2.Department of Interventional, the First Hospital Affiliated to Hebei North University, Zhangjiakou 075000, Hebei, China；3.School of Basic Medicine, Hebei North University, Zhangjiakou, Hebei 075000

摘要：

目的 探讨微小RNA-433(miR-433)靶向丝裂原活化蛋白激酶（MAPK）通路抑制胃癌BGC-823细胞增殖与侵袭的机制。方法 2021年10月至2022年5月期间，在河北北方学院附属第一医院胃肠肿瘤外科共收集30例胃癌组织及与之匹配的癌旁组织，以及人胃黏膜上皮细胞株GES-1及人胃癌细胞株BGC-823、MGC-803、SGC-7901。实时荧光定量PCR(qRT-PCR)检测胃癌组织、癌旁组织和GES-1、BGC-823、MGC-803、SGC-7901细胞中miR-433表达水平；对BGC-823细胞进行转染，分组为：空白组（细胞未转染）、miR-NC组（miR-433 mimics阴性对照转染细胞）、mi R-433模拟物（mimics）组（miR-433 mimics转染细胞），qRT-PCR检测各组BGC-823细胞miR-433表达水平；细胞计数试剂盒-8和Transwell实验分别检测各组BGC-823细胞增殖和侵袭能力；双荧光素酶报告基因实验验证mi R-433与MAPK靶向关系；蛋白印迹法检测各组BGC-823细胞中MAPK通路相关蛋白表达水平。结果 胃癌组织中miR-433表达水平显著低于癌旁组织（P<0.05）；与GES-1细胞比较，BGC-823、MGC-803、SGC-7901细胞中miR-433表达水平显著降低（P<0.05），且BGC-823细胞中mi R-433表达水平最低；与空白组和miR-NC组比较，miR-433 mimics组BGC-823细胞中mi R-433表达水平显著升高，细胞增殖率、侵袭细胞数、磷酸化p38 MAPK(p-p38 MAPK)/p38 MAPK、磷酸化JNK(p-JNK)/c-Jun氨基末端激酶（JNK）蛋白表达水平显著降低（P<0.05）；双荧光素酶报告基因实验结果 显示，miR-433与MAPK在BGC-823细胞中存在靶向关系。结论 上调miR-433表达可抑制胃癌BGC-823细胞的增殖与侵袭，该机制可能与靶向抑制MAPK通路有关。

关键词：微小RNA-433；丝裂原活化蛋白激酶；胃癌BGC-823细胞；增殖；侵袭

Abstract：

Objective To investigate the mechanism of microRNA-433 （miR-433） targeting mitogen activated protein kinase （MAPK） pathway to inhibit the proliferation and invasion of gastric cancer BGC-823 cells. Method From October 2021 to May 2022, the gastric cancer tissues and its matched paracancerous tissues from 30 patients in the Department of gastrointestinal tumor surgery, the First Hospital Affiliated to Hebei North University and human gastric mucosal epithelial cell line GES-1 and human gastric cancer cell lines BGC-823, MGC-803 and SGC-7901 were used. Real time fluorescent quantitative PCR （qRT-PCR） was used to detect the expression levels of miR-433 in gastric cancer tissues, paracancerous tissues and GES-1, BGC -823, MGC -803 and SGC -7901 cells; BGC -823 cells were transfected and divided into blank group（untransfected cells ）, miR-NC group （ miR - 433 mimics negative control transfected cells ） , miR – 433 mimics group （miR-433 mimics transfected cells）, the expression levels of miR-433 in BGC-823 cells was detected by qRT -PCR; cell counting kit -8 and Transwell assay were used to detect the proliferation and invasion of BGC -823 cells in each group; double luciferase reporter gene experiment verified the targeting relationship between miR -433 and MAPK; the expression levels of MAPK pathway related proteins in BGC -823 cells was detected by Western blot. Result The expression level of miR -433 in gastric cancer tissues was significantly lower than that in paracancerous tissues （P<0.05）; compared with GES-1 cells, the expression levels of miR -433 in BGC -823, MGC -803 and SGC -7901 cells was significantly decreased （P<0.05）, and the expression level of miR-433 in BGC-823 cells was the lowest; compared with the blank group and miR-NC group, the expression levels of miR -433 in BGC -823 cells in miR -433 mimics group was significantly increased, and the cell proliferation rate, the number of invasive cells, the expression levels of phosphorylated p38 MAPK （p -p38 MAPK）/p38 MAPK, phosphorylated JNK （p -JNK）/c -Jun amino terminal kinase （JNK） protein were significantly decreased （P<0.05）; the results of dual luciferase reporter gene experiments showed that miR-433 and MAPK were targeted in BGC-823 cells. Conclusion Up-regulation of miR -433 expression can inhibit the proliferation and invasion of gastric cancer BGC -823 cells, this mechanism may be related to the targeted inhibition of MAPK pathway.

Key Words: MicroRNA-433; Mitogen activated protein kinase; Gastric cancer BGC-823 cells; Proliferation; Invasion