体外转染JWA核酶逆转录病毒载体对人骨髓基质细胞黏附功能的影响  

Effect of transfected JWA ribozyme gene on adhension function of human bone marrow stromal cells in vitro

作　　者：常城[1] 陈幸华[1] 陆俊羽[1] 孔佩艳[1] 曾东风[1] 梁雪[1] 彭贤贵[1] 刘林[1] 刘红[1] 王庆余[1] 

机构地区：[1]第三军医大学新桥医院血液科,重庆400037

出　　处：《中华血液学杂志》2006年第12期805-808,共4页Chinese Journal of Hematology

摘　　要：目的探讨JWA核酶基因转染对人骨髓基质细胞(BMSC)黏附功能的影响。方法设计合成JWA核酶基因并构建于逆转录病毒载体pLXSN上,转染体外培养的人BMSC(BMSC-JWARZ组),应用半定量RT-PCR法(sqRT-PCR)测定细胞内JWA mRNA的表达,流式细胞仪检测BMSC细胞间黏附分子-1(ICAM-1)和血管黏附分子-1(VCAM-1)的表达,并检测共培养条件下BMSC对Jurkat细胞的黏附率及Jurkat细胞增殖变化。以pLXSN空载体转染的BMSC(BMSC-pLXSN组)及未转染细胞(BMSC组)为对照。结果BMSC-JWARZ组JWA mRNA表达量[吸光度(A)值]为0．187±0．045,较BMSC-pLXSN组(0．382±0．039)及BMSC组(0．366±0．048)显著降低;BMSC-JWARZ组细胞表面ICAM-1阳性细胞率为(16．11±3．93)％,较B-pLXSN组[(38．24±5．37)％]及BMSC组[(36．27±6．19)％]显著降低;BMSC-JWARZ组细胞表面VCAM-1阳性细胞率为(12．08±3．34)％,较BMSC- pLXSN组[(26．88±5．17)％]及BMSC组[(24．55±3．68)％]显著降低;B-JWARZ组细胞对Jurkat细胞的黏附率为(23．65±5．27)％,较BMSC-pLXSN组[(35．14±8．11)％]及BMSC组[(33．72±6．29)％]显著降低,与BMSC-JWARZ组细胞共培养的Jurkat细胞倍增时间为58．7 h,明显长于与BMSC-pLXSN组(44．1 h)及BMSC组(43．7 h)细胞共培养时。结论JWA核酶基凶转染可抑制人BMSC内JWA基因的表达,抑制骨髓基质细胞黏附功能。Objective To investigate the effects of transfected JWA ribozyme gene on adhension function of human bone marrow stromal cells （BMSC） in vitro. Methods JWA ribozyme gene was designed, synthesized and cloned into plasmid pLXSN, then the recombined plasmid was transferred into cultured human BMSC. Expression of JWA mRNA in transfected cells was detected by sqRT-PCR. Expressions of intercellular adhesion molecule-1 （ ICAM-1 ） protein and vascular adhesion molecule-1 （ VCAM-1 ） protein were detected by flow cytometry. Then the BMSC was co-cultured with Jurkat cells, and adhesion rate of BMSC to Jurkat cells and proliferation of Jurkat cells were measured. Cells transfected only with pLXSN and nontransfected cells were set as controls. Results The JWA mRNA expression（ A value） in cells transfected with JWA ribozyme gene （ B-JWARZ, 0. 187 ± 0. 045 ） was decreased significantly as compared with those in pLXSN transfected cells （ B-pLXSN, 0.382 ± 0. 039 ） and nontransfected control （ 0.366 ± 0. 048 ）. The expression of ICAM-1 protein, the expression of VCAM-1 protein and the adhesion rate to Jurkat cells all did the same as the JWA expression for the three cell groups. The population doubling times of Jurkat cells when co-cultured with B-JWARZ cells （58.7 h） decreased significantly than those when co-cultured with B-pLXSN group （44.1 h） and BMSC group（43.7 h）. Conchmion The JWA ribozyme gene transfection could inhibit the expression of JWA gene and reduce the adhesion functions of BMSC .

关 键 词：骨髓基质细胞 JURKAT细胞 黏附 JWA基因 核酶 

分 类 号：R733.71[医药卫生—肿瘤]