异丙酚通过miR-133a-3p/FTL轴调控GADD45A/JNK通路影响胃癌细胞增殖、迁移及凋亡  

Propofol regulates the GADD45A/JNK pathway through the miR-133a-3p/FTL axis and then affects the proliferation,migration and apoptosis of gastric cancer cells

作　　者：韦玲 杨凯 刘焕 程栋 WEI Ling;YANG Kai;LIU Huan;CHENG Dong(Department of Anesthesiology,Dongxihu District People’s Hospital,Wuhan 430040;Union Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430022,Hubei,China)

机构地区：[1]武汉市东西湖区人民医院麻醉科,湖北武汉430040 [2]华中科技大学同济医学院附属协和医院麻醉科,湖北武汉430022

出　　处：《川北医学院学报》2023年第8期1019-1025,共7页Journal of North Sichuan Medical College

基　　金：湖北省武汉市医学科研项目(WX20D18)。

摘　　要：目的:探讨异丙酚是否通过miR-133a-3p/FTL轴调控GADD45A/JNK通路影响胃癌细胞的增殖、迁移及凋亡。方法:将胃癌MKN-28细胞系分为磷酸盐缓冲液(PBS)组、异丙酚组、miR-NC组、miR-133a-3p mimics组、anti-miR-NC组、anti-miR-133a-3p组、si-NC组、si-FTL组、异丙酚+anti-miR-NC+si-NC组、PBS+anti-miR-NC+si-NC组、异丙酚+anti-miR-133a-3p+si-NC组、异丙酚+anti-miR-133a-3p+si-FTL组。应用RT-qPCR检测miR-133a-3p和铁蛋白轻链(FTL)mRNA的表达水平;应用MTT法与克隆形成实验测细胞增殖;应用流式细胞术、蛋白质印迹法分别检测细胞凋亡及蛋白表达水平;应用双荧光素酶报告实验检测miR-133a-3p与FTL的靶向关系;应用划痕实验与Transwell侵袭实验检测细胞迁移和侵袭。结果:与PBS组比较,异丙酚组细胞活性、划痕愈合率、细胞克隆形成数、迁移、侵袭细胞数均减少(P<0.05);细胞凋亡率、p53、Bcl-2相关X蛋白(Bax)、miR-133a-3p、GADD45A及p-JNK蛋白表达水平均升高(P<0.05),FTL表达水平降低(P<0.05)。双荧光素酶实验结果显示,miR-133a-3p靶向调控FTL。敲低FTL或过表达miR-133a-3p后,细胞凋亡率升高,细胞活性、划痕愈合率、细胞克隆形成数、迁移和侵袭细胞数降低;细胞中GADD45A、p-JNK、p53、Bax表达水平升高(P<0.05)。敲低FTL逆转了使用异丙酚及抑制miR-133a-3p表达对GADD45A/JNK信号通路以及MKN-28细胞恶性生物学行为的影响。结论:异丙酚可能通过miR-133a-3p/FTL轴调控GADD45A/JNK通路继而抑制胃癌细胞增殖、迁移,并促进细胞凋亡。Objective:To investigate whether propofol affects the proliferation,migration and apoptosis of gastric cancer cells by regulating the GADD45A/JNK pathway through miR-133a-3p/FTL axis.Methods:Gastric cancer cells MKN-28 were divided into phosphate buffered saline(PBS)group,propofol group,miR-NC group,miR-133a-3p mimics group,anti-miR-NC group,anti-miR-133a-3p group,si-NC group,si-FTL group,propofol+anti-miR-NC+si-NC group,PBS+anti-miR-NC+si-NC group,propofol+anti-miR-133a-3p+si-NC group,and propofol+anti-miR-133a-3p+si-FTL group.Real-time fluorescent quantitative PCR(RT-qPCR)was used to detect the expressions of miR-133a-3p and ferritin light chain(FTL)mRNA,cell proliferation was measured by MTT assay and clonal formation assay,apoptosis and protein expression were detected by flow cytometry and Western blot,dual luciferase reporter experiment was used to detect the targeting relationship between miR-133a-3p and FTL,Scratch assay and Transwell assay were used to detect cell migration and invasion.Results:Compared with PBS group,the cell activity,scratch healing rate,number of cell clone formation,migration and invasion cells in propofol group were decreased(P<0.05),the apoptosis rate,expression levels of p53,Bcl-2 related X protein(Bax),miR-133a-3p,GADD45A and p-JNK were increased(P<0.05),while FTL expression level was decreased(P<0.05).Dual luciferase experiment results showed that miR-133a-3p targeted regulation of FTL.After knockdown of FTL or overexpression of miR-133a-3p,cell apoptosis rate increased,cell activity,scratch healing rate,cell clone formation number,migration and invasion number decreased.The expression levels of GADD45A,p-JNK,p53 and Bax were increased(P<0.05).Knockdown of FTL reversed the effects of propofol and inhibiting the expression of miR-133a-3p on the GADD45A/JNK signaling pathway and the malignant biological behavior of MKN-28 cells.Conclusion:Propofol may regulate the GADD45A/JNK pathway through the miR-133a-3p/FTL axis,thereby inhibiting the proliferation and migration of gastric can

关 键 词：异丙酚 miR-133a-3p FTL GADD45A/JNK通路 胃癌 增殖 迁移 凋亡 

分 类 号：R735.2[医药卫生—肿瘤] R971[医药卫生—临床医学]