摘要
目的探讨去泛素化酶3(USP3)在子宫肌瘤中的表达情况以及其对子宫肌瘤细胞增殖及凋亡的影响。方法选择2017年4月至2018年7月子宫肌瘤切除手术患者56例。采用免疫组化、RT-PCR和Western blot检测USP3在子宫肌瘤组织、正常子宫肌层组织以及体外培养的子宫肌瘤细胞、子宫平滑肌细胞中的表达水平。设计合成USP3的干扰序列和对照序列,转染子宫肌瘤细胞,实验分为3组,即对照组、siRNA-Control组和siRNA-USP3组。Cell Counting Kit-8(CCK8)检测细胞增殖情况;流式细胞术观察细胞凋亡情况,Western blot检测细胞中p53和MDm2的表达水平。结果USP3在子宫肌瘤组织中的表达水平显著高于正常子宫肌层组织(P<0.05);USP3在子宫肌瘤细胞中的表达水平显著高于子宫平滑肌细胞(P<0.05)。敲低USP3后,子宫肌瘤细胞的增殖在24 h以后明显受到抑制;凋亡率显著增加;细胞中p53的表达显著升高(P<0.05);MDm2的表达显著降低(P<0.05)。结论USP3在子宫肌瘤中呈高表达,RNA干扰技术沉默USP3可以抑制子宫肌瘤细胞增殖,促进细胞凋亡,可能与抑制MDm2活性以及促进p53表达有关。
Objective To investigate the expression of ubiquitin-specific protease 3(USP3)in patients with uterine fibroids and its effects on cell proliferation and cell apoptosis of uterine fibroid cells.Methods A total of 56 patients with uterine fibroids who underwent hysteromyomectomy in our hospital from April 2017 to July 2018 were enrolled in the study.The expression levels of USP3 in uterine fibroids(observation group),myometrial tissue(control group)and in vitro cultured uterine fibroid cells and smooth muscle cells were detected by immunohistochemistry,reverse transcription-polymerase chain reaction(RT-PCR)and Western Blot,respectively.The interference sequence and control sequence of USP3 were designed and synthesized,then,which were transfected into uterine fibroid cells,which were divided into control group,siRNA-control group and siRNA-USP3 group.The expression levels of USP3 were detected by Western Blot,and the cell proliferation was tested by cell counting Kit-8(CCK8),moreover,the cell apoptosis was detected by flow cytometry,and the expression levels of p53 and mouse double minute 2(MDm2)were detected by Western Blot.Results The expression levels of USP3 in uterine fibroids were significantly higher than those in myometrial tissue(P<0.05).The expression levels of USP3 in uterine fibroid cells were significantly higher than those in smooth muscle cells(P<0.05).At 24 hours after knowing down the USP3,the cell proliferation was inhibited significantly,and the cell apoptosis was increased significantly,moreover,the expression levels of p53 were increased significantly(P<0.05),however,the expression levels of MDm2 were decreased significantly(P<0.05).Conclusion USP3 is highly expressed in uterine fibroids.Knocking down USP3 can inhibit the cell proliferation of uterine fibroid and promote cell apoptosis,its action mechanism may be correlated with inhibiting the activity of MDm2 and increasing the expression of p53.
作者
高光玲
李杨
杜玲莉
李珍珠
蒋海霞
GAO Guangling;Li Yang;DU Lingli(Guangyuan Hospital for Women and Children Health, Sichuan, Guangyuan 628000,China;不详)
出处
《河北医药》
CAS
2020年第13期1974-1978,共5页
Hebei Medical Journal
关键词
去泛素化酶3
子宫肌瘤
细胞增殖
细胞凋亡
ubiquitin-specific protease 3
uterine fibroids
cell proliferation
cell apoptosis
