KDR基因和Sema3基因在再障和正常人骨髓基质细胞中的表达

Expression of KDR and Sema3 genes in bone marrow stromal cells from patients with aplastic anemia and normal individuals

目的 :了解激酶插入区域受体 (kinaseinsertdomainreceptor,KDR)基因和脑信号蛋白Ⅲ(Semaphorin3,Sema3)基因在再生障碍性贫血 (AA)和正常人的骨髓基质细胞 (BMSC)和骨髓造血细胞中的表达情况。方法 :收集 9例AA和 33例正常骨髓标本 ,分离单个核细胞 (MNC)后体外长期培养扩增BMSC ,并收集悬浮细胞 (骨髓细胞 )。RT -PCR -ELISA检测KDR基因和Sema3基因在BM SC和造血细胞中的表达 ,分析表达率 ,并以管家基因 β2 微球蛋白 (β2 M)为内参照进行半定量分析。结果 :KDR基因在正常对照BMSC中的表达率 (97 0 % )明显高于对应的骨髓细胞 (70 8% ,P =0 0 0 7)。KDR基因在AA的BMSC和骨髓细胞中的表达率和表达水平与正常对照比较均无显著差异。Sema3基因在正常BMSC中的表达水平明显低于其对应的骨髓细胞 (P =0 0 35 )。Sema3基因在AA的BMSC和骨髓细胞中的表达率和表达水平与正常对照组比较均无显著差异。KDR基因和Sema3基因的表达水平在正常造血细胞中呈显著正直线相关 (r =0 70 3,P =0 0 0 2 )。结论 :KDR基因在BMSC中的高表达提示其可能在维持造血微环境中有重要作用。Sema3基因在造血细胞中的高表达状态提示其可能具有维持造血细胞生存的作用。

To investigate the expression of KDR (kinase insert domain receptor) and Sema3(semaphorin 3) genes in bone marrow stromal cells (BMSCs) and bone marrow hematopoietic cells from patients with AA (aplastic anemia) and normal individuals. Methods: Bone marrow mononuclear cells separated from AA (9 cases) and normal individuals (33 cases) were cultured in vitro. The adherent cells amplified by long-term culture and nonadherent cells were collected respectively. RT-PCR-ELISA assay was then performed to detect the expression of KDR and Sema3 genes. Expression ratios were analyzed and expression levels were normalized by housekeeping gene β_2 microglobulin (β_2M) acting as an internal calibration for the purpose of semi-quantitative analysis. Results: The expression ratio of KDR gene in BMSCs from normal control (97.0%) was higher than in its corresponding nonadherent cells (70.8%, P=0.007). There were no differences when expression ratios and levels of KDR gene in BMSCs and nonadherent cells from AA were compared with normal controls. Expression level of Sema3 gene in BMSCs from normal control was significantly lower than in its corresponding nonadherent cells (P=0.035). There were no differences when expression ratios and levels of Sema3 gene in BMSCs and nonadherent cells from AA were compared with normal controls. Expression levels of of Sema3 and KDR genes in the nonadherent cells from normal control display a significantly positive correlation (r=0.703, P=0.002). Conclusion: High expression of KDR gene in BMSCs suggests that it may play a critical role in sustaining the hematopoietic microenviroment. High expression of the Sema3 gene in hematopoietic cells suggests that it could sustain their survival. The significantly changes of KDR and Sema3 genes expression in AA could not be found.