miR-210-5p靶向Netrin-1在子痫前期中的表达及对绒毛外滋养层细胞增殖、凋亡、侵袭和迁移能力的影响  

Expression of miR-210-5p by targeting Netrin-1 in preeclampsia and its effect on proliferation,apoptosis,invasion and migration of extravillous trophoblast cells

作　　者：杜茜 雷利梅[1] DU Qian;LEI Limei(Department of Obstetrics and Gynecology,Mianyang Central Hospital affiliated to University of Electronic Science and Technology Mianyang Hospital,Mianyang,Sichuan 621000,China)

机构地区：[1]电子科学大学附属绵阳医院·绵阳市中心医院妇产科,四川绵阳621000

出　　处：《中国优生与遗传杂志》2023年第3期460-468,共9页Chinese Journal of Birth Health & Heredity

摘　　要：目的探究miR-210-5p靶向Netrin-1在子痫前期(PE)中的表达及对绒毛外滋养层细胞增殖、凋亡、侵袭和迁移能力的影响。方法选取2021年1月至12月在电子科学大学附属绵阳医院·绵阳市中心医院妇产科留存的62例PE患者和18例正常妊娠者的胎盘组织标本,免疫组化染色检测Netrin-1表达;RT-qPCR检测miR-210-5p、Netrin-1 mRNA表达。将HTR8-SVneo细胞进行不同转染,pHRi-anti-miR-NC、pHRi-anti-miR-210-5p、pHRi-NC、pHRi-Netrin-1分别转染至HTR8-SVneo细胞,pHRi-anti-miR-210-5p和pHRi-si-NC、pHRi-anti-miR-210-5p和pHRi-si-Netrin-1分别共转染至HTR8-SVneo细胞。双荧光素酶报告验证miR-210-5p对Netrin-1的调控关系;CCK-8及EdU染色、AnnexinV-FITC/PI法、划痕实验和Transwell实验分别检测细胞增殖、凋亡、迁移和侵袭能力;RT-qPCR检测miR-210-5p、Netrin-1 mRNA表达;Western blot检测Netrin-1、Ki67、Bcl2、Bax、MMP-2/9表达。结果与Control组相比,PE组miR-210-5p表达量升高,Netrin-1mRNA和蛋白表达量降低(均P<0.05)。miR-210-5p靶向负调控Netrin-1。转染pHRi-anti-miR-210-5p或pHRi-Netrin-1后,细胞24、48、72 h OD值及EdU阳性率升高,凋亡率降低,划痕愈合率和细胞侵袭数增加;Ki67、Bcl-2、MMP-2、MMP-9表达水平升高,Bax表达水平降低(均P<0.05);共转染pHRi-anti-miR-210-5p和pHRi-si-Netrin-1可减弱转染pHRi-anti-miR-210-5p对细胞生物学行为的作用。结论Netrin-1在PE中表达量降低,miR-210-5p表达量升高;抑制miR-210-5p表达可通过促进Netrin-1表达,促进绒毛外滋养层细胞增殖、侵袭和迁移能力,抑制其凋亡能力。Objective To explore the expression of miR-210-5p by targeting Netrin-1 in preeclampsia(PE)and its effect on proliferation,apoptosis,invasion and migration of extravillous trophoblast cells.Methods Placental tissue samples of 60 PE patients and 18 normal pregnant women in Department of Obstetrics and Gynecology in Mianyang Central Hospital affiliated to University of Electronic Science and Technology Mianyang Hospital from January 2021 to December 2021 were selected.Immunohistochemistry was used to detect the expression of Netrin-1.RT-qPCR was used to detect the expression of miR-210-5p and Netrin-1 mRNA.HTR8-SVneo cells were transfected differently,pHRi-anti-miR-NC,pHRi-anti-miR-210-5p,pHRi-NC,pHRiNetrin-1 were transfected into HTR8-SVneo cells respectively,and pHRi-anti-miR-210-5p and pHRi-si-NC,pHRi-anti-miR-210-5p and pHRi-si-Netrin-1 were co-transfected into HTR8-SVneo cells respectively.Double luciferase report was used to verify the regulatory relationship of miR-210-5p on Netrin-1.CCK-8 method and EdU staining,AnnexinV FITC/PI method,scratch test and Transwell test were used to detect the ability of proliferation,apoptosis,migration and invasion.RT-qPCR was used to detect the expression of miR-210-5p and Netrin-1 mRNA.Western blot was used to detect the expression of Netrin-1,Ki67,Bcl-2,Bax,MMP-2/9.Results Compared with the control group,the expression of miR-210-5p was increased and the expression of Netrin-1 mRNA was decreased(all P<0.05).miR-210-5p negatively regulated Netrin-1.After transfection of pHRi-anti-miR-210-5p or pHRi-Netrin-1,24,48,72 h OD value and EdU positive rate of cells were increased,the apoptosis rate was decreased,the scratch healing rate and the number of invasion cells were increased.The expression levels of Ki67,Bcl-2,MMP-2and MMP-9 were increased,while the expression levels of Bax was decreased(all P<0.05).Co-transfection of pHRi-anti-miR-210-5p and pHRi-si-Netrin-1 could weaken the effect of transfection of pHRi-anti-miR-210-5p on the biological behavior of cells.Conclusion The

关 键 词：miR-210-5p NETRIN-1 子痫前期 绒毛外滋养层细胞 

分 类 号：R714.244[医药卫生—妇产科学]