CacyBP/SIP对子宫内膜异位症间质细胞侵袭、迁移及血管生成的影响

Effects of CacyBP/SIP on Invasion,Migration and Angiogenesis of Stromal Cells in Endometriosis

目的探讨钙周期素结合蛋白(Calcyclin-Binding Protein/Siah-1-Interacting Protein,CacyBP/SIP)对子宫内膜异位症(Endometriosis,EMs)间质细胞侵袭、迁移及血管生成的作用机制。方法选取2021年1月至2022年12月在石家庄市妇幼保健院产科就诊的60例EMs患者为研究对象,经腹腔镜手术,取子宫内膜组织并分离子宫内膜间质细胞。使用重组CacyBP/SIP慢病毒、抑制剂转染细胞之后检测病毒、抑制剂转染结果,实验分为5组,分别为CacyBP/SIP慢病毒组、慢病毒阴性对照组、CacyBP/SIP抑制剂组、抑制剂阴性对照组及未治疗组。定量PCR鉴定各组细胞CacyBP/SIP水平,Transwell法检测各组细胞侵袭、迁移,MTT比色法检测各组细胞增殖,流式细胞仪检测各组细胞凋亡,免疫印迹法检测各组细胞血管内皮生长因子(Vascular Endothelial Growth Factor,VEGF)、环氧化酶-2(Cyclooxygenase-2,COX-2)表达水平。结果与抑制剂阴性对照组比较,CacyBP/SIP慢病毒组24、48、72 h细胞增殖升高(P<0.05),与CacyBP/SIP慢病毒组比较,CacyBP/SIP抑制剂组的细胞增殖降低(P<0.05);与抑制剂阴性对照组比较,CacyBP/SIP慢病毒组CacyBP/SIP水平、细胞侵袭数量、迁移数量及VEGF、COX-2蛋白表达升高(P<0.05),与CacyBP/SIP慢病毒组比较,CacyBP/SIP抑制剂组的上述指标降低(P<0.05);未治疗组、慢病毒阴性对照组、抑制剂阴性对照组、CacyBP/SIP慢病毒组、CacyBP/SIP抑制剂组细胞凋亡率分别为8.54%±0.26%、8.42%±0.38%、8.46%±0.19%、3.38%±0.15%、18.42%±1.23%,与抑制剂阴性对照组比较,CacyBP/SIP慢病毒组细胞凋亡率降低(P<0.05),与CacyBP/SIP慢病毒组比较,CacyBP/SIP抑制剂组细胞凋亡率升高(P<0.05)。结论CacyBP/SIP的低表达可降低EMs间质细胞的增殖、侵袭及迁移能力,推测其通过抑制VEGF、COX-2基因,可抑制血管生成。

Objective To explore the mechanism of Calcyclin-binding protein/Siah-1-interacting protein(CacyBP/SIP)on the invasion,migration,and angiogenesis of endometriosis(EMs)stromal cells.Methods A total of 60 patients with EMs who visited the Obstetrics Department of Shijiazhuang Maternal and Child Health Hospital from January 2021 to December 2022 were selected as the research subjects.Through laparoscopic surgery,endometrial tissue was taken and endometrial stromal cells were isolated.The recombinant CacyBP/SIP lentivirus and inhibitor transfection was used to detect the virus and inhibit the transfection results.The experiment was divided into 5 groups including CacyBP/SIP lentivirus group,lentivirus negative control group,CacyBP/SIP inhibitor group,inhibitor negative control group,and untreated group.Quantitative PCR was used to identify CacyBP/SIP levels in each group of cells,Transwell method was used to detect cell invasion and migration in each group,MTT colorimetric method was used to detect the proliferation of cells in each group,flow cytometry was used to detect cell apoptosis in each group,immunoblotting was used to detect the expression of vascular endothelial growth factor(VEGF)and cyclooxygenase-2(COX-2)in cells of each group.Results Compared with the inhibitor negative control group,the CacyBP/SIP lentivirus group showed an increase in cell proliferation at 24,48 and 72 h(P<0.05);however,compared with the CacyBP/SIP lentivirus group,the CacyBP/SIP inhibitor group showed a decrease in cell proliferation(P<0.05).Compared with the inhibitor negative control group,the CacyBP/SIP lentivirus group showed an increase in CacyBP/SIP levels,number of cell invasion,migration,and expression of VEGF and COX-2 proteins(P<0.05);however,compared with the CacyBP/SIP lentivirus group,the CacyBP/SIP inhibitor group showed a decrease in above indicators(P<0.05).The cell apoptosis rates in the untreated group,lentivirus negative control group,inhibitor negative control group,CacyBP/SIP lentivirus group,and CacyBP/SIP inhibitor group were 8.54%±0.26%,8.42%±0.38%,8.46%±0.19%,3.38%±0.15%,and 18.42%±1.23%,respectively.Compared with the inhibitor negative control group,the cell apoptosis rate in the CacyBP/SIP lentivirus group decreased(P<0.05);compared with the CacyBP/SIP lentivirus group,the apoptosis rate of cells in the CacyBP/SIP inhibitor group increased(P<0.05).Conclusion The low expression of CacyBP/SIP can reduce the proliferation,invasion and migration ability of EMs mesenchymal cells,which is speculated to inhibit angiogenesis by inhibiting VEGF and COX-2 genes.