OXT通过miR-196b-3p/ASPP2信号通路调节卵巢癌细胞生长  

OXT Regulates Ovarian Cancer Cell Growth via miR-196b-3p/ASPP2 Signal Pathway

作　　者：吴莉敏 陈茜[1] 赵艳[1] 吴莉娜[2] 冯晓珊 沈梦丹 马文娟[1] 孙艳萍 WU Li-min;CHEN Qian;ZHAO Yan;WU Li-na;FENG Xiao-shan;SHEN Meng-dan;MA Wen-juan;SUN Yan-ping(Department of Gynaecology and Obstetrics,the Second Affiliated Hospital of Xi’an Jiaotong University,Xi’an 710004,China;Operating Theater,the Second Affiliated Hospital of Xi’an Jiaotong University,Xi’an 710004,China)

机构地区：[1]西安交通大学第二附属医院妇产科,西安710004 [2]西安交通大学第二附属医院手术室,西安710004

出　　处：《南昌大学学报（医学版）》2020年第2期53-60,共8页Journal of Nanchang University：Medical Sciences

摘　　要：目的探讨催产素(oxytocin,OXT)对卵巢癌细胞增殖与凋亡的影响及其作用机制。方法人卵巢癌细胞系SKOV3和A2780细胞来源于美国模式培养物保藏所(ATCC)。OXT和OXT受体(oxytocin receptor,OXTR)拮抗剂Atosiban刺激卵巢癌细胞,改变细胞中miR-196b-3p的水平,使用MTT法、ELISA试验、Western blot、生物信息学工具及荧光素酶报告基因实验等研究以上处理对卵巢癌细胞增殖与凋亡的作用和机制。结果OXT刺激SKOV3细胞OXTR激活(F=28.842,P<0.05),增殖活性降低(F=12.988,P<0.05),Caspase3活性增加(F=26.676,P<0.05);Ki67、增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)及p53凋亡刺激蛋白抑制剂(inhibitor of apoptosis-stimulating of p53 protein,iASPP)表达下调,p53凋亡刺激蛋白(apoptosis-stimulating of p53 protein,ASPP)1(ASPP1)及2(ASPP2)表达上调;但Atosiban部分逆转OXT的效果。OXT刺激卵巢癌细胞miR-196b-3p水平减少(A2780:F=76.406,P<0.05;SKOV3:F=45.874,P<0.05)。miR-196b-3p-mimic转染联合OXT刺激,导致细胞增殖活性增加(F=9.232,P<0.05),Caspase3活性降低(F=36.350,P<0.05),同时ASPP2表达降低(F=83.013,P<0.05)。而miR-196b-3p-inhibitor转染联合OXT刺激,ASPP2表达增加(F=83.013,P<0.05)。生物信息学工具预测联合荧光素酶报告基因实验确认ASPP2是miR-196b-3p的靶基因。结论OXT介导的OXTR激活通过调节miR-196b-3p/ASPP2信号通路调节卵巢癌细胞生长。Objective To investigate the effect of oxytocin(OXT) on proliferation and apoptosis of ovarian cancer cells and its mechanism of action.Methods The ovarian cancer cell lines SKOV3 and A2780 were purchased from American Type Culture Collection. Cells were stimulated by OXT alone or in combination with OXT receptor(OXTR) antagonist Atosiban. The miR-196 b-3 p mimic and inhibitor were used to change the levels of miR-196 b-3 p in the cells. MTT method,ELISA assay,Western blot,bioinformatics tools and luciferase reporter assay were used to confirm the effect and mechanism of OXT.Results After stimulation with OXT alone,the OXTR was activated(F=28.842,P<0.05),the proliferation was inhibited(F=12.988,P<0.05),caspase-3 activity was increased(F=26.676,P<0.05),the expression of ki-67,proliferating cell nuclear antigen and inhibitor of apoptosis-stimulating of p53 protein(ASPP) was down-regulated,and the expression of ASPP1 and ASPP2 was up-regulated in SKOV3 cells. However,the effects of OXT were partly reversed by Atosiban.The levels of miR-196 b-3 p were decreased after OXT stimulation in both A2780(F=76.406,P<0.05) and SKOV3 cells(F=45.874,P<0.05). After miR-196 b-3 p-mimic transfection,the proliferation was enhanced(F=9.232,P<0.05),caspase-3 activity was suppressed(F=36.350,P<0.05),and ASPP2 expression was decreased(F=83.013,P<0.05) in OXT-treated cells. Nevertheless,the expression of ASPP2 was increased after transfection with miR-196 b-3 p-inhibitor(F=83.013,P<0.05). Bioinformatics prediction and luciferase reporter gene experiment confirmed that ASPP2 was a direct target gene of miR-196 b-3 p.Conclusion OXT-mediated OXTR activation regulates ovarian cancer cell growth via miR-196 b-3 p/ASPP2 signaling pathway.

关 键 词：催产素 卵巢癌 细胞增殖 细胞凋亡 microRNA-196b-3p p53凋亡刺激蛋白2 

分 类 号：R34[医药卫生—基础医学] R737.32