胃癌间质干细胞活化Wnt/β-catenin信号通路促进胃癌细胞增殖及迁移作用的探讨  

Gastric cancer-derived mesenchymal stem cells promote gastric cancer cell proliferation and migration by activating Wnt/β-catenin signaling pathway

作　　者：尹磊 刘伟 胡玉燕 蒋涵 钱晖 YIN Lei;LIU Wei;HU Yuyan;JIANG Han;QIAN Hui(Jiangsu Key Institute of Clinical Laboratory Medicine,Jiangsu University Medical College,Zhenjiang 212013,Jiangsu,China)

机构地区：[1]江苏大学医学院&检验医学江苏省重点实验室,江苏镇江212013

出　　处：《临床检验杂志》2020年第5期371-375,共5页Chinese Journal of Clinical Laboratory Science

基　　金：镇江市外泌体基础与转化应用高技术研究重点实验室(SS2018003);江苏省普通高校研究生科研创新计划项目(KYCX17_1819);江苏大学大学生科研课题资助项目(18A498)。

摘　　要：目的探讨胃癌间质干细胞(gastric-cancer-derived mesenchymal stem cells,GC-MSCs)活化Wnt/β-catenin信号通路促进胃癌细胞增殖、迁移的作用。方法采用贴壁培养法从胃癌组织中分离GC-MSCs,成骨、成脂培养基诱导GC-MSCs分化。收集GC-MSC上清,胃癌细胞系HGC-27分别经L-DMEM培养液(Control组)、GC-MSC上清(GC-MSC组)、GC-MSC上清及20μmol/L ICG001(GC-MSC+ICG001组)处理24 h,克隆形成试验和Transwell迁移试验分别检测细胞增殖、迁移能力,免疫荧光染色或western blot检测Wnt/β-catenin信号相关蛋白(β-catenin、CD44、CyclinD3)的表达水平。结果 GC-MSC上清处理组HGC-27细胞增殖形成的克隆数[(203.700±8.762)个]显著高于Control组[(33.670±1.856)个](t=18.98,P<0.01),且其迁移细胞数[(349.700±7.881)个]较Control组[(145.000±3.712)个]亦显著增多(t=23.46,P<0.01)。GC-MSC上清可促进HGC-27细胞β-catenin入核,诱导Wnt/β-catenin信号下游蛋白CD44、CyclinD3的表达。Wnt/β-catenin信号抑制剂ICG001可下调上述分子的表达,逆转GC-MSC上清对HGC-27细胞的促增殖、促迁移作用。与GC-MSC组[(203.000±3.606)个]相比,GC-MSC+ICG001组的克隆细胞数[(70.667±2.603)个]显著减少(q=39.24,P<0.01),且迁移细胞数[(166.000±3.215)个]较GC-MSC组[(352.700±4.096)个]显著降低(q=47.73,P<0.01)。结论 GC-MSCs活化Wnt/β-catenin信号并促进胃癌细胞增殖及迁移。Objective To investigate the role of gastric-cancer-derived mesenchymal stem cells(GC-MSCs)in the activation of Wnt/β-catenin signaling pathway and promotion of gastric cancer cell proliferation and migration.Methods GC-MSCs were isolated from gastric cancer tissues by adherent culture,and the differentiation of GC-MSCs was induced by osteogenic and lipogenic media.GC-MSC supernatant was collected and the gastric cancer cell HGC-27 were treated with L-DMEM medium(control group),GC-MSC supernatant(GC-MSC group)and GC-MSC supernatant combined with 20μmol/L ICG001(GC-MSC+ICG001 group)for 24 hours,respectively.The cell proliferation and migration were detected by clone formation and Transwell migration assays,respectively.The expression levels of Wnt/β-catenin signal related proteins(β-catenin,CD44,Cyclin D3)were analyzed by immunofluorescence or western blot.Results The number of clones(203.700±8.762)formed by HGC-27 cell proliferation in GC-MSC supernatant-treated group was significantly higher than that in control group(33.670±1.856)(t=18.98,P<0.01),and the number of migrated cells(349.700±7.881)was significantly higher than that in control group(145.000±3.712)(t=23.46,P<0.01).GC-MSC supernatant promoted HGC-27β-catenin nuclear transport and induced the expressions of CD44 and CyclinD3,the downstream proteins of Wnt/β-catenin signal pathway.ICG001,an inhibitor of Wnt/β-catenin signal reduced the expression of these molecules and reversed the promoting effects of GC-MSC supernatant on the proliferation and migration of HGC-27 cells.Compared with GC-MSC group(203.000±3.606),the number of clones in GC-MSC+ICG001 group(70.667±2.603)was significantly reduced(q=39.24,P<0.01)and the number of migrated cells in GC-MSC+ICG001 group(166.000±3.215)was significantly lower than that of GC-MSC group(352.700±4.096,q=47.73,P<0.01).Conclusion GC-MSCs could activate Wnt/β-catenin signal and promote the proliferation and migration of gastric cancer cells.

关 键 词：胃癌间质干细胞 胃癌 增殖 迁移 WNT/Β-CATENIN 

分 类 号：R735.2[医药卫生—肿瘤]