摘要
目的miR-24靶向EGFR基因对幽门螺杆菌感染的胃癌细胞生长和迁移的影响。方法体外培养幽门螺杆菌感染胃癌细胞MGC803,将miR-NC、miR-24、si-NC、si-EGFR、miR-24+pcDNA-NC和miR-24+pcDNA-EGFR转染至MGC803细胞。实时荧光定量聚合酶链反应(RT-qPCR)检测细胞miR-24和EGFR mRNA表达;细胞计数试剂盒(CCK-8)检测细胞生长;Transwell小室实验检测细胞迁移;蛋白质印迹检测细胞周期素D1(CyclinD1)、基质金属蛋白酶2(MMP-2)蛋白表达。双荧光素酶报告实验验证miR-24与EGFR的靶向结合关系。结果与正常对照组比较,幽门螺杆菌感染组胃癌细胞miR-24表达降低,EGFR mRNA表达升高(P<0.05)。与miR-NC组比较,miR-24组细胞miR-24表达升高,细胞存活率、迁移细胞数以及CyclinD1蛋白、MMP-2蛋白表达降低(P<0.05)。与si-NC组比较,si-EGFR组细胞EGFR mRNA表达降低,细胞存活率、迁移细胞数以及CyclinD1蛋白、MMP-2蛋白表达降低(P<0.05)。与miR-24+pcDNA-NC组比较,miR-24+pcDNA-EGFR组细胞存活率、迁移细胞数以及CyclinD1蛋白、MMP-2蛋白表达升高(P<0.05)。结论miR-24通过抑制EGFR表达降低幽门螺杆菌感染的胃癌细胞生长和迁移。
OBJECTIVE To investigate the effect of miR-24 targeting EGFR gene on the growth and migration of gastric cancer cells infected by Helicobacter pylori.METHODS H.pylori was cultured in vitro to infect gastric cancer cells MGC803,and miR-NC,miR-24,si-NC,si-EGFR,miR-24+pcDNA-NC and miR-24+pcDNA-EGFR were transfected into MGC803 cells.Real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)was used to detect the expression of miR-24 and EGFR mRNA in cells;cell counting kit(CCK-8)was used to detect cell growth.Cell migration was detected by transwell chamber experiment.Western blot was used to detect protein expression of cyclin D1(CyclinD1),matrix metalloproteinase 2(MMP-2).The dual luciferase reporter experiment was used to verify the targeted binding relationship between miR-24 and EGFR.RESULTS Compared with the normal control group,the expression of miR-24 in gastric cancer cells of the H.pylori infection group was significantly decreased,and the expression of EGFR mRNA was significantly increased(P<0.05).Compared with the miR-NC group,the expression of miR-24 in the miR-24 group was significantly increased,while the cell survival rate,the number of migrating cells,and the expression of CyclinD1 and MMP-2 proteins were significantly decreased(P<0.05).Compared with the si-NC group,the EGFR mRNA expression,the cell survival rate,the number of migrating cells,and the expression of CyclinD1 protein and MMP-2 protein in cells of the si-EGFR group were significantly reduced(P<0.05).Compared with the miR-24+pcDNA-NC group,the cell survival rate,the number of migrating cells and the expression of CyclinD1 protein and MMP-2 protein in the miR-24+pcDNA-EGFR group were significantly increased(P<0.05).CONCLUSION miR-24 can reduce the growth and migration of gastric cancer cells infected by H.pylori by inhibiting the expression of EGFR.
作者
张红
宋志雪
孙新光
朱笑葳
唐梦宇
陈长香
ZHANG Hong;SONG Zhi-xue;SUNXin-guang;ZHU Xiao-wei;TANG Meng-yu;CHEN Chang-xiang(Tianjin Tianshi College,Tianjin 301700,China;不详)
出处
《中华医院感染学杂志》
CAS
CSCD
北大核心
2022年第23期3587-3591,共5页
Chinese Journal of Nosocomiology
基金
天津市教委科研计划基金资助项目(2020SK138)。
