SEMA3F通过CXCR4/JAK2/STAT3通路抑制子宫内膜癌细胞增殖、侵袭、迁移及血管生成  

SEMA3F suppresses the proliferation,invasion,migration and angiogenesis of endometrial cancer cells via CXCR4/JAK2/STAT3 pathway

作　　者：时周云 陈红晶 钱琳玉 朱玲玲[1] SHI Zhouyun;CHEN Hongjing;QIAN Linyu;ZHU Lingling(Department of Obstetrics and Gynecology,Nantong Maternal and Child Health Hospital,Jiangsu Nantong 226018,China)

机构地区：[1]南通市妇幼保健院妇产科,江苏南通226018

出　　处：《现代肿瘤医学》2023年第7期1218-1225,共8页Journal of Modern Oncology

基　　金：江苏省南通市科技计划(指导性)项目(编号:JCZ20022)。

摘　　要：目的:探究信号素3F(semaphorin-3F,SEMA3F)通过趋化因子受体4(C-X-C motif chemokine receptor 4,CXCR4)/JANUS激酶2(janus kinase 2,JAK2)/信号转导及转录激活蛋白3(signal transducer and activator of transcription 3,STAT3)通路对子宫内膜癌细胞增殖、侵袭、迁移及血管生成的调控作用。方法:体外培养人子宫内膜基质细胞系、人子宫内膜癌细胞(Ishikawa、KLE、RL95-2、HEC-1-A)及人脐静脉血管内皮细胞(human umbilical vein endothelial cells,HUVEC)。构建含有SEMA3F重组质粒的慢病毒载体并转染至Ishikawa细胞,分为对照组(Control)、过表达对照组(Ov-NC)、SEMA3F过表达组(Ov-SEMA3F);将含SEMA3F重组质粒的慢病毒载体与构建的含CXCR4重组质粒的慢病毒载体或STAT3激动剂(Colivelin)共转染至Ishikawa细胞,分为4组:Control组、Ov-SEMA3F组、Ov-SEMA3F+Ov-CXCR4组、Ov-SEMA3F+Colivelin组。实时荧光定量PCR(quantitative real-time PCR,RT-qPCR)、蛋白质印迹法(Western blot)检测SEMA3F、CXCR4表达水平;EdU染色检测细胞增殖;划痕和Transwell实验分别检测细胞迁移和侵袭能力;Transwell和小管形成实验分别检测Ishikawa细胞上清作用下HUVEC迁移和子宫内膜癌细胞小管形成能力;Western blot分析增殖和转移相关蛋白及CXCR4/JAK2/STAT3通路相关蛋白表达水平。结果:SEMA3F在子宫内膜癌细胞中表达降低(均P<0.01),SEMA3F过表达可抑制细胞增殖、迁移、侵袭和血管生成(均P<0.001)。SEMA3F过表达可抑制CXCR4/JAK2/STAT3信号通路(P<0.001),CXCR4过表达或STAT3激动剂(Colivelin)可逆转SEMA3F对Ishikawa细胞增殖、迁移、侵袭及Ishikawa细胞上清作用下HUVEC细胞迁移、血管生成能力的抑制作用(均P<0.05)。结论:SEMA3F可通过使CXCR4/JAK2/STAT3通路失活进而抑制子宫内膜癌细胞增殖、侵袭、迁移及血管生成。Objective:To figure out the regulatory role of SEMA3F in the proliferation,invasion,migration and angiogenesis of endometrial cancer cells via C-X-C motif chemokine receptor 4(CXCR4)/Janus kinase 2(JAK2)/signal transducer and activator of transcription 3(STAT3)pathway.Methods:Human endometrial stromal cell line,endometrial cancer cells(Ishikawa,KLE,RL95-2 and HEC-1-A)and human umbilical vein endothelial cells(HUVEC)were cultured in vitro.Lentiviral vector containing SEMA3F recombinant plasmid was constructed and transfected into Ishikawa cells which were divided into Control group,Ov-NC group and Ov-SEMA3F group.Lentiviral vector containing SEMA3F recombinant plasmid were co-transfected with constructed lentiviral vector containing CXCR4 recombinant plasmid or STAT3 activator Colivelin into Ishikawa cells which were divided into 4 groups:Control group,Ov-SEMA3F group,Ov-SEMA3F+Ov-CXCR4 group and Ov-SEMA3F+Colivelin group.RT-qPCR and Western blot detected the expression levels of SEMA3F and CXCR4.Cell proliferation was estimated by EdU staining.Cell migration and invasion capacities were respectively measured through wound healing and Transwell assays.Transwell and tube formation assays respectively detected the migration and tube formation of HUVEC in Ishikawa cells supernatant.Western blot tested the expression levels of proliferation-,metastasis-associated proteins and CXCR4/JAK2/STAT3 signaling-associated factors.Results:SEMA3F expression was reduced in endometrial cancer cells(all P<0.01)and SEMA3F overexpression suppressed the proliferation,migration,invasion and angiogenesis of endometrial cancer cells(all P<0.001).SEMA3F overexpression also inhibited CXCR4/JAK2/STAT3 pathway(P<0.001).Further,CXCR4 overexpression or STAT3 activator Colivelin reversed the inhibitory role of SEMA3F in Ishikawa cell proliferation,migration,invasion,as well as HUVEC migration and angiogenesis in Ishikawa cells supernatant(all P<0.05).Conclusion:SEMA3F might inactivate CXCR4/JAK2/STAT3 pathway to further inhibit the proliferatio

关 键 词：子宫内膜癌 SEMA3F 血管生成 CXCR4/JAK2/STAT3通路 

分 类 号：R737.33[医药卫生—肿瘤]