骨髓间充质干细胞来源外泌体通过ERK1/2信号通路调控子痫前期滋养层细胞侵袭和增殖相关机制研究  

The Mechanism of Human Bone Mesenchyma Stem Cell Derived Exosomes on Invasion and Proliferation of Trophoblast Cells in Preeclampsia Through ERK1/2 Signal Pathway

作　　者：宋素香 徐琳[1] SONG Suxiang;XU Lin(Affiliated Hospital of Qingdao University,Qingdao,266000)

机构地区：[1]青岛大学附属医院,266000

出　　处：《实用癌症杂志》2023年第7期1045-1050,共6页The Practical Journal of Cancer

摘　　要：目的探究骨髓间充质干细胞(human bone mesenchyma stem cell,BMSCs)来源外泌体通过ERK1/2信号通路调控子痫前期滋养层细胞的增殖和侵袭的机制。方法以大鼠来源的BMSCs为种子细胞,采用超速离心法进行收集BMSCs来源外泌体。将子痫前期滋养层细胞分成3组,即实验组,抑制剂组和对照组,其中实验组滋养层细胞与外泌体共培养,抑制剂组滋养层细胞与外泌体共培养后,加入ERK1/2的抑制剂AZD0364,对照组滋养层细胞与PBS溶液共培养。应用CCK-8试剂盒和Transwell试剂盒检测细胞的增殖和侵袭能力。应用RT-PCR和Western blot检测各组细胞的ERK1/2和核因子NF-κB的相对表达量。结果透射电镜结果显示,BMSCs来源外泌体为直径50~150 nm的双层膜囊泡状结构。外泌体与子痫前期滋养层细胞共培养的过程中,可见MEG-01细胞可以摄取并内化PKH67标记的外泌体。CCK-8试剂盒结果显示,实验组细胞增殖率要明显高于对照组,而抑制剂组细胞的增殖率明显低于实验组(P<0.05)。Transwell试剂盒检测结果显示,实验组细胞的侵袭明显强于对照组(P<0.05),抑制剂组细胞的侵袭明显弱于实验组(P<0.05)。RT-PCR和Western blot结果显示,实验组ERK1/2和NF-κB的相对表达量明显高于对照组(P<0.05),抑制剂组ERK1/2和NF-κB的相对表达量明显低于实验组(P<0.05)。结论BMSCs来源外泌体可以提高ERK1/2信号通路表达,进而促进子痫前期滋养层细胞的增殖和侵袭。Objective To explore the mechanism of extramesenchymal stem cell proliferation and the regulation of bone marrow stromal secretion in preeclampsia.Methods The BMSCs from rats were used as seed cells,and the exosomes from BMSCs were collected by ultracentrifugation.The trophoblast cells of preeclampsia were divided into 3 groups:the experimental group,the inhibitor group and the control group.The trophoblast cells of the experimental group were co cultured with exosomes.After the trophoblast cells of the inhibitor group were co cultured with exosomes,AZD0364,an inhibitor of ERK1/2,was added.The trophoblast cells of the control group were co cultured with PBS solution.The CCK-8 kit and the Transwell kit were used to detect the proliferation and invasion of cells.The ERK1/2 and nuclear factor NF-κB were detected by RT-PCR and Western blot.Results The reslut of the transmission electron microscopy showed that the secretes from BMSCs were bilayer vesicles with a diameter of 50-150 nm.During the co-culture of exosomes and pre-eclampsia trophoblast cells,the MEG-01 cells can absorb and internalize PKH67 labeled exosomes.The results of CCK-8 kit showed that the proliferation rate of cells in the experimental group was significantly higher than that in the control group,while the proliferation rate of cells in the inhibitor group was significantly lower than that in the experimental group(P<0.05).The results of Transwell kit showed that the invasion of cells in the experimental group was significantly higher than that in the control group(P<0.05),and the invasion of cells in the inhibitor group was significantly lower than that in the experimental group(P<0.05).The results of the RT-PCR showed that the relative expression of ERK1/2 and NF-κB in the inhibitor group was significantly higher than that in the control group(P<0.05),The relative expression of the relative expression of ERK1/2 and NF-κB was significantly lower than that in the experimental group(P<0.05).Conclusion BMSCs derived exosomes can increase the expre

关 键 词：间充质干细胞 外泌体 子痫前期 增殖 侵袭 

分 类 号：R733.3[医药卫生—肿瘤]