miR-195通过靶向调控FOXK1对胃癌细胞增殖、侵袭及转移的影响  

Mechanism of miR-195 regulating FOXK1 over the proliferation,invasion and metastasis of gastric cancer cell

作　　者：薛兰辉 冯谢敏[1] 宋炜[1] Xue Lanhui;Feng Xiemin;Song Wei(Ward I,Department of Oncology,Yan'an University Affiliated Hospital,Yan'an 716000,China)

机构地区：[1]延安大学附属医院肿瘤科一病区,延安716000

出　　处：《中国实用医刊》2024年第17期1-5,共5页Chinese Journal of Practical Medicine

摘　　要：目的探究微小RNA-195(miR-195)通过靶向调控叉头框蛋白K1(FOXK1)对胃癌细胞增殖、侵袭及转移的影响。方法体外培养胃癌MGC803细胞,将miR-195或miR-NC mimics、inhibitor对照序列通过Lipofectamine2000转染试剂转染入细胞内,并依次将其命名为miR-195 mimics组、mimics-NC组、miR-195 inhibitor组和inhibitor-NC组。采用逆转录聚合酶链反应(RT-PCR)检测细胞中miR-195 mRNA表达,蛋白免疫印迹(Western blot)方式检测FOXK1蛋白具体表达情况,采用CCK8方式检测细胞的具体增殖水平,细胞的具体侵袭能力分析采用Tranwell实验,细胞的具体转移能力采用划痕试验予以检测,miR-195与FOXK1靶向关系验证则采用荧光素酶实验。结果与inhibitor-NC组相比,miR-195 inhibitor组miR-195 mRNA表达降低,细胞增殖水平、Tranwell实验穿膜数、迁移距离、FOXK1蛋白表达均增大(P均<0.05);与mimics-NC组相比,miR-195 mimics组miR-195 mRNA表达升高,细胞增殖水平、Tranwell实验穿膜数、迁移距离、FOXK1蛋白表达均降低(P均<0.05);荧光素酶实验显示,miR-195可直接靶向作用于FOXK1蛋白。结论miR-195可抑制胃癌细胞增殖、侵袭和转移,可能通过调控FOXK1表达而起作用。Objective To investigate the effect of microRNA-195(miR-195)regulating Forkhead box k1(FOXK1)over the proliferation,invasion and metastasis of gastric cancer.Methods Gastric cancer MGC803 cells were cultured in vitro,and miR-195 or miR-NC mimics and inhibitor control sequences were transfected into the cells through Lipofectamine 2000 transfection reagent,and they were named in sequence as the miR-195 mimics group,mimics-NC group,miR-195 inhibitor group and inhibitor-NC group.Reverse transcription-polymerase chain reaction was used to detect the expression of miR-195 mRNA in cells;the specific expression of FOXK1 protein was detected by Western blot;the specific proliferation level of cells was detected using Cell Counting Kit-8 method;the specific invasion ability of cells was analyzed using Tranwell assay;and the specific metastasis ability of cells was detected by scratch test.The luciferase assay was used to verify the targeting relationship between miR-195 and FOXK1.Results Compared with the inhibitor-NC group,the expression of miR-195 mRNA in the miR-195 inhibitor group decreased,moreover,the cell proliferation level,number of membrane penetrations in Tranwell assay,migration distance and FOXK1 protein expression all increased in the miR-195 inhibitor group(all P<0.05).Compared with the mimics-NC group,the expression of miR-195 mRNA increased in the-195 mimics group,while the cell proliferation level,number of membrane penetrations in Tranwell assay,migration distance and FOXK1 protein expression all decreased in the miR-195 mimics group(all P<0.05).The luciferase assay showed that miR-195 can directly target on FOXK1 protein.Conclusions MiR-195 can inhibit the proliferation,metastasis and invasion of gastric cancer cells,which may act by regulating the expression of FOXK1.

关 键 词：胃癌 微小RNA-195 叉头框蛋白K1 增殖 侵袭转移 

分 类 号：R735.2[医药卫生—肿瘤]