jagged1活化notch通路促进raw264.7向破骨分化但抑制增殖①

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1、王汝杰等Jagged1活化Notch通路促进ＲAW264.7向破骨分化但抑制增殖第7期·865·doi:10．3969/j．issn．1000-484X．2014．07．001·基础免疫学·①Jagged1活化Notch通路促进ＲAW264.7向破骨分化但抑制增殖王汝杰刘复州沈伟伟胡旭陈培毛德举卓云云陈武桂周跃初同伟(第三军医大学第二附属医院骨科，重庆400037)中图分类号Ｒ392文献标志码A文章编号1000-484X(2014)07-0865-05［摘要］目的:探讨Jagged1通过活化Notch通路对ＲAW264.7细胞向破骨分化及增殖的影响。方法:将ＲAW2

2、64.7细胞分三组培养:对照组(细胞培养基+鼠ＲANKL50ng/L)、Jagged1组(细胞培养基+鼠ＲANKL+Jagged1重组蛋白)、γ-分泌酶抑制剂(DAPT)组(细胞培养基+鼠ＲANKL+Jagged1重组蛋白+DAPT)，实时荧光定量PCＲ检测各组破骨细胞标志基因组织蛋白酶K(CathepsinK，CK)、抗酒石酸性磷酸酶(Tartrate-resistantacidphosphatase，TＲAP)、降钙素受体(Calcitioninre-ceptor，CTＲ)及Notch靶基因HES-1、HEY-1mＲNA的表达，TＲAP染色鉴定破骨细胞，扫描电镜

3、检测破骨细胞溶骨功能。免疫ICD荧光检测N的表达变化，CCK-8检测ＲAW264.7细胞增殖。结果:Jagged1组TＲAP、CK、CTＲ及HES-1、HEY-1mＲNA的表+达、TＲAP细胞数较对照组及DAPT组明显升高(P＜0.05)，而DAPT组与对照组均无明显变化;细胞免疫荧光显示Jagged1ICDICD组N除表达于细胞膜、细胞质外，细胞核也有较高表达，对照组及DAPT组细胞核中N无明显表达;细胞培养48h，Jagged1组细增殖较对照组及DAPT组出现明显抑制。结论:Jagged1通过活化Notch通路促进ＲAW264.7向破骨细胞分化、抑制其增殖。［关

4、键词］Jagged1;破骨细胞;Notch;细胞分化;细胞增殖Jagged1promotesosteoclastdifferentiationandinhibitsproliferationbyactiva-tingofNotchsignalingpathwayinducedWANGＲu-Jie，LIUFu-Zhou，SHENWei-Wei，HUXu，CHENPei，MAODe-Ju，ZHUOYun-Yun，CHENWu-Gui，ZHOUYue，CHUTong-Wei.DepartmentofOrthopedics，SecondAffiliatedHospital，

5、ThirdMilitaryMedicalUniversity，Chongqing400037，China［Abstract］Objective:TostudytheroleofJagged1andNotchsignalingpathwayplayedinthedifferentiationandproliferationofＲAW264.7cells.Methods:ＲAW264.7cellsweredividedintothreegroupstoculture:Thecontrolgroup:ＲAW264.7cellswerethreatedwithculture

6、andＲANKL.TheJagged1group:ＲAW264.7cellswerethreatedwithrecombinantproteinJagged1besidesthecontrolgroup.TheDAPTgroup:ＲAW264.7cellswerethreatedwithDAPTbesidestheJagged1group.ThemＲNAexpressionofosteo-clastmarkers(TＲAP，CK，CTＲ)andNotchkeytargetgenes(HES-1andHEY-1)weremeasuredbyreal-timePCＲ.T

7、heformationofosteoclast，boneresorption，NotchexpressionandproliferationofＲAW264.7cellsweredetectedbyTＲAPstaining，scanningelectronmicroscope，immunofluorescenceandcellcountingkit-8(CCK-8).Ｒesults:TＲAP，CK，CTＲ，HES-1andHEY-1mＲNAexpression+weresignificantlyhigherthanthecontrolgroupandDAPTgr