Caspase-3和GSDME介导的细胞焦亡对子宫肌瘤细胞侵袭和迁移的影响  

Effects of Caspase-3 and GSDME-mediated pyroptosis on invasion and migration of uterine fibroids

作　　者：谌媛媛 胡银逢 刘金龙[2] SHEN Yuanyuan;HU Yinfeng;LIU Jinlong(Obstetrics and Gynecology Department of the 1 st Affiliated Hospital of Anhui Medical University,Hefei 230022,Anhui,China;Neurosurgery Department of Linquan County People's Hospital,Linquan 236400,Anhui,China)

机构地区：[1]安徽省医科大学第一附属医院妇产科,安徽合肥230022 [2]临泉县人民医院神经外科,安徽临泉236400

出　　处：《贵州医科大学学报》2022年第9期1037-1043,共7页Journal of Guizhou Medical University

基　　金：安徽省卫生计生委科研计划项目(2016QK033)。

摘　　要：目的 探讨Caspase-3/消皮素E(GSDME)介导的细胞焦亡对子宫肌瘤细胞侵袭和迁移的影响。方法 采用实时荧光定量PCR(qRT-PCR)检测子宫平滑肌细胞及子宫肌瘤细胞中caspase-3和GSDME mRNA表达;构建GSDME过表达质粒并转染至子宫肌瘤细胞,再将细胞分为空载体质粒组(Vector组)和转染GSDME载体质粒组(OVE-GSDME组),采用Western blot法检测GSDME、GSDME-N、caspase-3及cleaved-caspase-3蛋白表达水平,采用酶联免疫法检测乳酸脱氢酶(LDH)含量,显微镜下观察细胞的形态,Transwell法检测细胞的侵袭和迁移能力。结果 子宫肌瘤细胞caspase-3和GSDME mRNA表达水平低于子宫平滑肌细胞(P<0.01),OVE-GSDME组子宫肌瘤细胞的GSDME、GSDME-N、caspase-3、cleaved-caspase-3蛋白表达水平及LDH含量高于Vector组细胞(P<0.01),并且OVE-GSDME组子宫肌瘤细胞的体积增大、并出现“吹泡”形状,表现出典型的细胞焦亡现象;Transwell结果显示,OVE-GSDME组子宫肌瘤细胞的细胞侵袭和迁移指数低于Vector组(P<0.01)。结论 激活Caspase-3/GSDME,可能通过介导细胞焦亡的机制抑制子宫肌瘤细胞侵袭和迁移能力。Objective To investigate the effect of Caspase-3/GSDME-mediated pyroptosis on the invasion and migration of uterine fibroids.Methods Real-time fluorescence quantitative PCR(qRT-PCR)was used to detect the expression of caspase-3 and GSDME mRNA in uterine smooth muscle cells and uterine fibroids cells.GSDME overexpression plasmids were constructed and transfected into uterine fibroids.The experiments were divided into transfection blank plasmid group(Vector group)and transfected GSDME plasmid group(OVE-GSDME group).The protein expressions of GSDME,GSDME-N,caspase-3 and cleaved-caspase-3 were detected by Western blotting;lactate dehydrogenase(LDH)release was detected by ELISA,cell morphology was observed by microscope,and cell invasion and migration were detected by Transwell.Results Lower expression of caspase-3 and GSDME mRNA was found in uterine fibroids than uterine smooth muscle cells(P<0.01).GSDME,GSDME-N,caspase-3,and cleaved-caspase-3 protein expression and LDH release were more promoted than Vector group(P<0.01).Uterine fibroids cellular size was enlarged,bubbling form and typical cell pyroptosis were observed in OVE-GSDME group.Transwell experiments showed that uterine fibroids cell of OVE-GSDME group showed less cell invasion and migration indices than Vector gorup(P<0.01).Conclusion Activation of Caspase-3/GSDME can inhibit invasion and migration of uterine fibroids cell by mediating pyroptosis mechanism.

关 键 词：子宫肌瘤 CASPASE-3 消皮素E(GSDME) 焦亡 侵袭 迁移 细胞 

分 类 号：R737[医药卫生—肿瘤]