CAJ | 학술논문

目的观察Jagged1通过活化Notch通路对人外周血CD14+单核细胞破骨分化、增殖的影响.方法免疫组织化学法检测Jagged1在肺癌骨转移标本中的表达,Ficoll法联合磁珠分选法分离人外周血CD14+单核细胞,分别加入Jagged1重组蛋白、Jagged1重组蛋白及γ-分泌酶抑制剂(DAPT)分组培养,采用实时定量聚合酶链反应(Real-time PCR)检测组织蛋白酶K(CK)、抗酒石酸性磷酸酶(TRAP)、降钙素受体(CTR)及Notch靶基因发状分裂相关增强子-1(HES-1)、HES相关YRPW主题-1(HEY-1)mRNA的表达,TRAP染色鉴定破骨细胞,扫描电镜检测破骨细胞溶骨功能.细胞计数法(CCK-8)检测CD14+单核细胞增殖.结果 Jagged1在肺癌骨转移标本中高表达,癌旁组织无或低表达;Jagged1组TRAP、CK、CTR及HES-1、HEY-1 mRNA的表达、TRAP+细胞数较对照组及DAPT组明显升高(P＜0.05),而DAPT组与对照组比较均无明显变化;细胞培养48 h,Jagged1组细胞增殖较对照组及DAPT组明显抑制.结论 Jagged1通过活化Notch通路促进破骨前体细胞分化、抑制其增殖.
목적 관찰Jagged1통과활화Notch통로대인외주혈CD14+단핵세포파골분화、증식적영향.방법 면역조직화학법검측Jagged1재폐암골전이표본중적표체,Ficoll법연합자주분선법분리인외주혈CD14+단핵세포,분별가입Jagged1중조단백、Jagged1중조단백급γ-분비매억제제(DAPT)분조배양,채용실시정량취합매련반응(Real-time PCR)검측조직단백매K(CK)、항주석산성린산매(TRAP)、강개소수체(CTR)급Notch파기인발상분렬상관증강자-1(HES-1)、HES상관YRPW주제-1(HEY-1)mRNA적표체,TRAP염색감정파골세포,소묘전경검측파골세포용골공능.세포계수법(CCK-8)검측CD14+단핵세포증식.결과 Jagged1재폐암골전이표본중고표체,암방조직무혹저표체;Jagged1조TRAP、CK、CTR급HES-1、HEY-1 mRNA적표체、TRAP+세포수교대조조급DAPT조명현승고(P＜0.05),이DAPT조여대조조비교균무명현변화;세포배양48 h,Jagged1조세포증식교대조조급DAPT조명현억제.결론 Jagged1통과활화Notch통로촉진파골전체세포분화、억제기증식.
Objective To study the role of Jadded1 and Notch signaling pathway played in the differentiation and proliferation of osteoclasts derived from human circulating CD14 + monocytes.Methods Peripheral blood CD14 + monocytes were isolated from healthy adult by Ficoll density gradient centrifugation combined with magnetic beads separation.Recombinant protein Jagged1 and γ-Secretase Inhibitor (DAPT) with Jagged1 were added to the culture system of CD14 + monocytes respectively.The mRNA expression of osteoclast markers tartrate-resistant acid phosphatase (TRAP),cathepsin K (CK),calctionin receptor (CTR) and Notch key target genes hairy and enhancer of split-1 (HES-1) and HES-related with YRPW motif-1 (HEY-1) were measured by quantitative real-time quantitative polymerase chain reaction (Real-time PCR).The formation of osteoclast,bone resorption,Notch expression and proliferation of CD14 + monocytes were detected by TRAP staining,scanning electron microscope,and cell counting kit-8 (CCK-8).Results High expression of Jagged1 in bone metastases specimens was observed in contrast of no expression or low expression in adjacent tissues.TRAP,CK,CTR,HES-1 and HEY-1 mRNA expression were significantly higher than the control group and DAPT group in Jadded1 group (P ＜ 0.05).TRAP + cell count,osteolytic area was significantly increased in Jagged1 group when compared with control and DAPT group,and no significant difference observed between the last two groups.Cell proliferation was inhibited in Jagged1 group also (P ＜ 0.05).Conclusion Jagged1 promotes osteoclast differentiation and inhibits proliferation by activating Notch signaling pathway.