摘要
目的探讨长链非编码RNA LINC00662调节miR-497-5p/活化T细胞核因子5(NFAT5)轴对子宫内膜癌(EC)细胞增殖、迁移及侵袭的影响。方法qRT-PCR检测人正常子宫内膜上皮细胞HUEEpiC和人EC细胞AN3CA、Ishikawa、HEC-1A中LINC00662、miR-497-5p、NFAT5 mRNA表达水平。将AN3CA细胞分为Control组、sh-NC组、sh-LINC00662组、sh-LINC00662+anti-NC组、sh-LINC00662+anti-miR-497-5p组,检测各组细胞中LINC00662、miR-497-5p、NFAT5 mRNA和蛋白质的表达及细胞的增殖、迁移、侵袭、凋亡,双荧光素酶实验验证LINC00662与miR-497-5p、miR-497-5p与NFAT5的关系。结果人EC细胞中,LINC00662、NFAT5 mRNA高表达,miR-497-5p低表达,且AN3CA细胞中LINC00662、miR-497-5p、NFAT5 mRNA表达差异最大,选取AN3CA细胞进行实验。双荧光素酶实验显示,LINC00662与miR-497-5p、miR-497-5p与NFAT5具有靶向结合关系。与Control组或sh-NC组相比,sh-LINC00662组细胞的存活率、克隆数、PCNA、MMP-2、MMP-9、划痕愈合率、侵袭数,以及LINC00662、NFAT5 mRNA和蛋白质表达降低,细胞凋亡率、miR-497-5p表达升高(P<0.05);与sh-LINC00662组或sh-LINC00662+anti-NC组相比,sh-LINC00662+anti-miR-497-5p组存活率、克隆数、PCNA、MMP-2、MMP-9、划痕愈合率、侵袭数、NFAT5 mRNA和蛋白质表达升高,细胞凋亡率、miR-497-5p表达降低(P<0.05)。结论人EC细胞中,LINC00662高表达,敲低LINC00662可能通过调控miR-497-5p/NFAT5轴,抑制人EC细胞AN3CA的增殖、迁移及侵袭。
Objective To investigate the effects of long non coding RNA LINCO0662 on the proliferation,migration,and invasion of endometrial cancer(EC)cells by regulating the miR-497-5p/nuclear factor of activated T cells 5(NFAT5)axis.Methods qRT-PCR was used to detect the expression levels of LINC00662,miR-497-5p,and NFAT5 mRNA in human normal endometrial epithelial cells HUEEpiC and human EC cells AN3CA,Ishikawa,HEC-1A.AN3CA cells were divided into Control group,sh-NC group,sh-LINC00662 group,sh-LINC00662+anti-NC group,and sh-LINC00662+anti-miR-497-5p group.The mRNA and protein expressions of LINC00662,miR-497-5p and NFAT5 in each group were detected,and the cell proliferation,migration,invasion and apoptosis were detected.Dual luciferase assay was used to detect the relationship between LINC00662 and miR-497-5p,as well as miR-497-5p and NFAT5.Results Human EC cells had high expression of LINC00662 and NFAT5 mRNA and low expression of miR-497-5p,and AN3CA cells had the greatest difference in the expression of LINC00662,miR-497-5p and NFAT5 mRNA.AN3CA cells were selected for experiments.Dual luciferase assay showed that LINC00662 had targeted binding with miR-497-5p,and miR-497-5p had targeted binding with NFAT5.Compared with the Control group or sh-NC group,the survival rate,clone number,PCNA,MMP-2,MMP-9,scratch healing rate,invasion number,LINC00662,NFAT5 mRNA and protein expression were lower in the sh-LINC00662 group,the apoptosis rate and miR-497-5p expression were higher(P<0.05).Compared with the sh-LINC00662 group or sh-LINC00662+anti-NC group,the survival rate,clone number,PCNA,MMP-2,MMP-9,scratch healing rate,invasion number,LINC00662,NFAT5 mRNA and protein expression were higher in the sh-LINC00662+anti-miR-497-5p group,the apoptosis rate and miR-497-5p expression were lower(P<0.05).Conclusion LINC00662 is highly expressed in human EC cells,and knocking down LINC00662 may inhibit the proliferation,migration,and invasion of human EC cells AN3CA by regulating the miR-497-5p/NFAT5 axis.
作者
张梦
马雪燕
梁楠楠
ZHANG Meng;MA Xueyan;LIANG Nannan(Department of Gynecology,Tangshan Maternal and Children Health Hospital,Tangshan,Hebei 063000,China)
出处
《中国优生与遗传杂志》
2025年第3期545-552,共8页
Chinese Journal of Birth Health & Heredity
