摘要
目的:探讨环状RNA浆细胞瘤变体易位1(Hsa-circ-PVT1)对子宫腺肌病异位子宫内膜间质细胞增殖、侵袭和血管生成的影响,以及其对微小RNA145(miR-145)和丝裂原活化蛋白激酶1(MAPK1)基因的调控机制。方法:qRT-PCR检测子宫腺肌病异位子宫内膜中Hsa-circ-PVT1、miR-145表达。双荧光素酶实验检测Hsa-circ-PVT1和miR-145的关系。生物信息数据库TargetScanHuman、miRDB、miRactDB预测miR-145的下游靶点。细胞分组:Hsa-circ-PVT1-inhibitor-NC+miR-145-antagomir-NC+si-MAPK1-NC(NC)、Hsa-circ-PVT1-inhibitor(inhibitor)、Hsa-circ-PVT1-inhibitor+miR-145-antagomir(inhibitor+antagomir)、Hsa-circ-PVT1-inhibitor+miR-145-miR-145-antagomir+si-MAPK1(inhibitor+antagomir+si)、正常细胞(normal组)。EDU检测各组细胞增殖,Transwell小室检测各组细胞侵袭能力;小管形成实验检测各组细胞血管生成能力;Western blot法检测各组细胞中MAPK1、增殖细胞核抗原(PCNA)、基质金属蛋白酶-9(MMP-9)和血管内皮生长因子(VEGF)表达水平。结果:子宫腺肌病异位子宫内膜中Hsa-circ-PVT1表达明显升高,miR-145表达明显下降,双荧光素酶证实Hsa-circ-PVT1靶向miR-145的表达,miR-145下游靶标为MAPK1。与normal组Y14细胞相比,inhibitor组细胞增殖、侵袭及血管新生的性能明显降低,inhibitor+antagomir组能部分逆转这一抑制作用,而inhibitor+antagomir+si组能部分恢复这一作用,差异均有统计意义(均P<0.05)。结论:子宫腺肌病异位子宫内膜中Hsa-circ-PVT1表达明显升高,Hsa-circ-PVT1能靶向miR-145调控MAPK1表达,抑制Hsa-circ-PVT1表达能明显抑制间质细胞的增殖、侵袭及血管生成。
Objective:To investigate the effect of cyclic RNA plasmacytoma variant translocation 1(Hsa-circ-PVT1)on the cell proliferation,invasion and angiogenesis of ectopic endometrial stromal cells of adenomyosis(ADS),and its regulation mechanism on microRNA145(miR-145)and mitogen-activated protein kinase 1(MAPK1).Methods:qRT-PCR was used to detect the expressions of Hsa-circ-PVT1 and miR-145 in clinical tissue samples.The dual luciferase assay was used to detect the relationship between Hsa-circ-PVT1 and miR-145.Bioinformatics databases TargetScanHuman,miRDB and miRactDB predict the downstream targets of miR-145.The cell groups were as follows:Hsa-circ-PVT1-inhibitor-NC+miR-145-antagomir-NC+si-MAPK1-NC(NC),Hsa-circ-PVT1-inhibitor(inhibitor),Hsa-circ-PVT1-inhibitor+miR-145-antagomir(inhibitor+antagomir),Hsa-circ-PVT1-inhibitor+miR-145-miR-145-antagomir+si-MAPK1(inhibitor+antagomir+si),and normal cells(nomal group)were taken separately.EDU was used to detect the proliferation activity of each group of cells.Transwell chamber was used to detect the invasion ability of each group of cells.The tubule formation experiment was used to detect the angiogenesis ability of each group of cells.Western blot was used to detect the expression levels of MAPK1 and proliferating cell nuclear antigen(PCNA),matrix metalloproteinases 9(MMP-9)and vascular endothelial growth factor(VEGF).Results:The expression of Hsa-circ-PVT1 in the ectopic endometrium of adenomyosis was significantly increased,the expression of miR-145 and was significantly decreased.Double luciferase confirmed that Hsa-circ-PVT1 targeted the expression of miR-145,and the downstream target of miR-145 was MAPK1.Compared with normal group,the proliferation,invasion and angiogenesis of Y14 cells in inhibitor group were significantly decreased.Inhibitor+antagomir group could partially reverse this inhibitory effect,while inhibitor+antagomir+si group could partially restore this effect,the difference was statistically significant(all P<0.05).Conclusion:In the ectopic endometrium
作者
陈慧
郎茂竹
王婧
朱亚莉
田艳
Chen Hui;Lang Maozhu;Wang Jing(Department of Gynecology,Affiliated Hospital of Guizhou Medical University,Guiyang 550004)
出处
《现代妇产科进展》
北大核心
2023年第3期182-187,共6页
Progress in Obstetrics and Gynecology
