EGF促进人羊膜间充质干细胞迁移上调MMP14和IL-1β表达  

EGF promotes migration and expressions of MMP14 and IL-1β in human amniotic mesenchymal stem cells

作　　者：谭丽萍[1] 庞希宁[1] 施萍[2] 王竟[3] 王喜良[1] 

机构地区：[1]中国医科大学 卫生部细胞生物学重点实验室 干细胞与再生医学研究室,辽宁沈阳110001 [2]中国医科大学附属第一医院全科医学教研室,辽宁沈阳110001 [3]中国医科大学附属第一医院肛肠外科,辽宁沈阳110001

出　　处：《基础医学与临床》2013年第11期1404-1409,共6页Basic and Clinical Medicine

基　　金：国家重点基础研究发展计划(2012CB518103);国家自然科学基金(81370883);辽宁省科学技术计划(2012225080);沈阳市科学技术计划(F11-262-9-01)

摘　　要：目的研究表皮细胞生长因子(EGF)促进人羊膜间充质干细胞(hAMSCs)迁移过程对基质金属蛋白酶14(MMP14)和白细胞介素1β(IL-1β)表达的影响。探讨EGF促进hAMSCs迁移的机制。方法贴壁法分离培养原代细胞流式细胞术鉴定Transwell小室检测EGF对hAMSCs迁移能力影响;Western blot和real-time PCR检测MMP14和IL-1β蛋白及mRNA表达。结果原代细胞表达间充质干细胞表面标记CD44,不表达CD45;不同浓度EGF作用hAMSCs 24 h 100μg/L EGF迁移指数最大为2.0;100μg/L EGF作用hAMSCs12、24和48 h迁移指数为1.6、2.0和1.7;MMP14蛋白表达12 h为(0.81±0.20)显著高于对照组的(0.63±0.12)(P<0.05);IL-1β蛋白表达48 h为(0.55±0.16)显著高于对照组的(0.30±0.05)(P<0.05)MMP14和IL-1βmRNA表达上调显著高于对照组(P<0.05)。结论 EGF促进hAMSCs迁移上调MMP14和IL-1β表达;MMP14、IL-1β和EGF协同激活MAPK/ERK、P13K/AKT信号通路参与调控EGF促进hAMSCs迁移。Objective To research the expression regulation of epidermal growth factor （EGF） on matrix metallo- proteinase 14 and interleukin-1β in the progress of human amniotic mesenehymal stem cell migration and explore the mechanisms of EGF to promote hAMSCs migration. Methods Take health cesarean cultured amniotic mem- branes adherent mesenchymal stem cells, using transwell cell migration in vitro system of observation EGF on migra- tion of hAMSCs ; By Western blot detection EGF action hAMSCs different time MMP14 and IL-1β protein expression changes, Using real-time quantitative PCR （Real-time PCR ） detection of EGF action hAMSCs different time MMPI4 and IL-1β expression levels changes of mRNA. Results Primary isolated and cultured human amniotic） cells Identified by flow cytometry express surface marker of mesenchymal stem cells CD44 do not express CD45, tran-swell chamber detect different concentration of EGF acts on hAMSCs 24 h reveal that 100 ~g/L EGF is the best： 2. 0,100 μg/L EGF acts on hAMSCsl2, 24 and 48 h, migration index namely 1.6,2. 0 and 1.7;The protein level of MMP14 and IL-1β were markedly increased by 100 ng/mL EGF 12 h results of MMP14 are（0. 81 ± 0. 20）signif- icantly higher than that of control group （0. 63± 0. 12） ;the 48 h results of IL-1β are （0. 55±0. 16 ）significantly higher than control group（0. 30 ±0. 05 ）, 100 μg/L EGF upregulated the express of MMP14 and IL-1β mRNA sig- nificantly higher than control group（P 〈 0. 05 ）. Conclusions EGF has a significant role in promoting the migration of hAMSCs.

关 键 词：基质金属蛋白酶14 白细胞介素1Β 表皮生长因子 羊膜间充质干细胞 细胞迁移 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学]