miRNA-34b对子宫内膜癌细胞凋亡、侵袭、迁移的影响及作用机制  

Effect and Mechanism of miR-34b on Apoptosis,Invasion and Migration of Endometrial Cancer Cells

作　　者：李晓妍 高秀娟[2] 刘雪洁 LI Xiao-yan;GAO Xiu-juan;LIU Xue-jie(Department of Gynaecology,the Second Central Hospital of Baoding,Hebei Baoding 072750,China;Department of Gynecology,Affiliated Hospital of Chengde Medical College,Hebei Chengde 067020,China)

机构地区：[1]保定市第二中心医院妇科,河北保定072750 [2]承德医学院附属医院妇科,河北承德067020

出　　处：《现代检验医学杂志》2022年第6期88-93,98,共7页Journal of Modern Laboratory Medicine

基　　金：河北省医学科学研究课题(编号:20200265),资助单位:河北省卫生健康委员会。

摘　　要：目的探究miR-34b对子宫内膜癌(endometrial cancer)凋亡、迁移、侵袭的影响及对Jag1/Cyclin D1信号通路的影响。方法将人子宫内膜癌细胞AN3CA细胞分为对照组、NC组和miR-34b组。通过流式细胞技术、Transwell实验分别检测各组细胞凋亡、迁移能力和侵袭能力;通过定量聚合酶链反应(quantitative polymerase chain reaction,qPCR)及Westernblot检测各组细胞Jagged-1蛋白(Jag1)及其靶基因G1/S-特异性周期蛋白-D1(Cycling D1)mRNA和蛋白的表达水平;通过裸鼠荷瘤实验分析miR-34b对肿瘤生长的影响。结果对照组、NC组和miR-34b组细胞凋亡比例分别为5.6%±0.12%,5.80%±0.22%和19.4%±0.51%。miR-34组细胞凋亡比例高于对照组,差异具有统计学意义(t=8.325,P<0.01);对照组、NC组和miR-34b组细胞迁移数分别为322.15±13.71个,327.67±9.14个和162.19±7.49个,miR-34b组细胞迁移数低于对照组,差异具有统计学意义(t=7.945,P<0.01);对照组、NC组和miR-34b组细胞侵袭数分别为357.6±14.11个,364.05±16.12个和157.58±8.77个,miR-34b组细胞侵袭数低于对照组,差异具有统计学意义(t=7.643,P<0.01);qPCR结果表明对照组、NC组和miR-34b组Jag1 mRNA表达为2.75±0.21,2.67±0.14和1.19±0.19,miR-34b组低于对照组,差异具有统计学意义(t=8.434,P<0.01);对照组、NC组和miR-34b组蛋白表达为1.71±0.11,1.77±0.24和0.69±0.16,miR-34b组低于对照组,差异具有统计学意义(t=7.895,P<0.01);对照组、NC组和miR-34b组Cyclin D1mRNA表达为1.29±0.13,1.32±0.11和0.59±0.13,miR-34b组低于对照组,差异具有统计学意义(t=9.124,P<0.01);对照组、NC组和miR-34b组蛋白表达为1.81±0.18,1.79±0.14和1.29±0.16,miR-34b组低于对照组,差异具有统计学意义(t=8.227,P<0.01);裸鼠荷瘤实验表明过表达miR-34b裸鼠肿瘤生长曲线与对照组差异有统计学显著性意义,并且肿瘤体积显著低于对照组(t=8.184,P<0.01)。结论miR-34b可以促进人子宫内膜癌细胞的凋亡,抑制细胞迁移和侵袭Objective To investigate the effect and mechanism of miR-34b on apoptosis,migration and invasion of endometrial cancer cells.Methods Human endometrial cancer cell line AN3CA was divided into control group,NC group and miR-34b group.The apoptosis,migration and invasion of each group were detected by flow cytometry and Transwell experiment respectively.The mRNA and protein expression levels of Notch ligand(Jag1)and its target gene G1/S-specific cyclin-D1 were detected by qPCR and Western blot.The effect of miR-34b on tumor growth was analyzed by tumor bearing experiment in nude mice.Results The apoptosis rates of control group,NC group and miR-34b group were 5.6%±0.12%,5.80%±0.22%and 19.4%±0.51%,respectively.The proportion of apoptosis in miR-34 group was higher than that in control group,the difference was statistically significant(t=8.325,P<0.01).The number of cell migration in the control group,NC group and miR-34b group were 322.15±13.71,327.67±9.14 and 162.19±7.49,respectively.The number of cell migration in miR-34b group was lower than that in the control group,and the difference was statistically significant(t=7.945,P<0.01).The number of cell invasion in the light group,NC group and miR-34b group were 357.6±14.11,364.05±16.12 and157.58±8.77,respectively,and the number of cell invasion in miR-34b group was lower than that in the control group,and the difference was statistically significant(t=7.643,P<0.01).QPCR results showed that the expression of Jag1 mRNA in control group,NC group and miR-34b group was 2.75±0.21,2.67±0.14 and 1.19±0.19,respectively.The expression of Jag1 mRNA in miR-34b group was lower than that in control group,and the difference was statistically significant(t=8.434,P<0.01).The protein expression in control group,NC group and miR-34b group was 1.71±0.11,1.77±0.24 and 0.69±0.16,respectively.The expression of Jag1 mRNA in miR-34b group was lower than that in control group,and the difference was statistically significant(t=7.895,P<0.01).The expression of cyclin D1 mRNA in con

关 键 词：miR-34b 人子宫内膜癌 凋亡 迁移与侵袭 Jag1/Cyclin D1信号通路 

分 类 号：R737.33[医药卫生—肿瘤] R730.43[医药卫生—临床医学]