摘要
目的 探究缺氧条件下胆管癌(cholangiocarcinoma,CCA)细胞分泌的外泌体(exosome,Exo)对CCA细胞的增殖、迁移、侵袭及Janus激酶2(Janus kinase 2,JAK2)/信号转导和转录激活因子3(signal transducer and activator of transcription 3,STAT3)通路的影响。方法 提取CCA细胞分泌的Exo并鉴定。将常氧、缺氧条件下的Exo与CCA细胞共培养,分为常氧组、缺氧组、缺氧+抑制剂组,另设空白组,检测CCA细胞的增殖、迁移、侵袭能力和相关蛋白表达。结果 在CCA细胞质中检测到Exo携带荧光染料3,3'-双十八烷基氧杂碳菁高氯酸盐。与空白组比较,常氧组的克隆形成数、细胞迁移数、侵袭细胞数量上升,增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)、基质金属蛋白酶(matrix metallopeptidase,MMP)-2、MMP-9、神经钙黏蛋白(neural cadherin,N-Cadherin)、波形蛋白(Vimentin)、兔抗磷酸化JAK2(phosphorylated JAK2,p-JAK2)/JAK2、兔抗磷酸化STAT3(phosphorylated STAT3,p-STAT3)/STAT3表达升高,上皮细胞钙黏蛋白(epithelial cadherin,E-Cadherin)表达降低(P<0.05);与常氧组比较,缺氧组的克隆形成数、细胞迁移数、侵袭细胞数量上升,PCNA、MMP-2、MMP-9、N-Cadherin、Vimentin、p-JAK2/JAK2、p-STAT3/STAT3表达升高,E-Cadherin表达降低(P<0.05);与缺氧组比较,缺氧+抑制剂组的克隆形成数、细胞迁移数、侵袭细胞数量下降,PCNA、MMP-2、MMP-9、N-Cadherin、Vimentin、p-JAK2/JAK2、p-STAT3/STAT3表达降低,E-Cadherin表达升高(P<0.05)。结论 缺氧条件下CCA细胞分泌的Exo通过激活JAK2/STAT3通路促进CCA细胞的增殖、迁移、侵袭及上皮间质转化。
Objective To investigate the effects of exosome(Exo)secreted by cholangiocarcinoma(CCA)cells under hypoxic conditions on the proliferation,migration,invasion,and Janus kinase 2(JAK2)/signal transducer and activator of transcription 3(STAT3)pathway of CCA cells.Methods Exo secreted by CCA cells was extracted and identified.Exo under normoxic and hypoxic conditions were co-cultured with CCA cells and divided into normal oxygen group,hypoxic group,hypoxic+inhibitor group,and blank group.The proliferation,migration,and invasion abilities of CCA cells were tested separately.Immunoblotting was applied to detect the expression levels of relevant proteins.Results Exo carrying fluorescent dye 3,3'-dioctadecyloxacarbocyanine perchlorate was detected in CCA cytoplasm.Compared with blank group,the number of clone formation,cell migration and invasion cells in normoxic group increased,the expression of proliferating cell nuclear antigen(PCNA),matrix metallopeptidase(MMP)-2,MMP-9,neural cadherin(N-Cadherin),Vimentin,phosphorylated JAK2(p-JAK2)/JAK2,phosphorylated STAT3(p-STAT3)/STAT3 increased,and the expression of epithelial cadherin(E-Cadherin)decreased(P<0.05);Compared with normal oxygen group,the number of clone formation,cell migration and invasion cells in hypoxia group increased,the expression of PCNA,MMP-2,MMP-9,N-Cadherin,Vimentin,p-JAK2/JAK2 and p-STAT3/STAT3 increased,and the expression of E-Cadherin decreased(P<0.05).Compared with hypoxia group,the number of clone formation,cell migration and invasion cells in hypoxia+inhibitor group decreased,the expression of PCNA,MMP-2,MMP-9,N-Cadherin,Vimentin,p-JAK2/JAK2 and p-STAT3/STAT3 decreased,and the expression of E-Cadherin increased(P<0.05).Conclusion Exo secreted by CCA cells under hypoxic conditions promote proliferation,migration,invasion,and epithelial mesenchymal transition of CCA cell by activating the JAK2/STAT3 pathway.
作者
张剑华
苏子剑
连锦鸿
ZHANG Jianhua;SU Zijian;LIAN Jinhong(Department of Hepatobiliary Surgery,Quanzhou First Hospital,Quanzhou 362000,Fujian,China)
出处
《中国现代医生》
2025年第24期29-34,共6页
China Modern Doctor
基金
福建医科大学启航项目(2020QH1270)。
关键词
缺氧
胆管癌
外泌体
增殖
迁移
侵袭
Janus激酶2/信号转导和转录激活因子3通路
Hypoxia
Cholangiocarcinoma
Exosome
Proliferation
Migration
Invasion
Janus kinase 2/signal transducer and activator of transcription 3 pathway
