骨髓间质干细胞与肿瘤细胞中FN1、NME2、TIMP3基因表达检测

Detection for expression level of gene FN1,NME2 and TIMP3 in mesenchymal stem cells,tumor cells and leukemia cells

目的探讨纤维连接蛋白1(FN1)、肿瘤转移抑制基因2(NME2)、组织金属蛋白酶抑制因子3(TIMP3)基因在胎儿骨髓间质干细胞(FMSCs)、成人骨髓间质干细胞(AMSCs)、F6、SGC及白血病细胞中的差异表达,寻找肿瘤和白血病的分子生物学特征。方法分别提取FMSCs、AMSCs、F6、SGC及白血病细胞的总RNA,逆转录cDNA,以cDNA为模板用PCR和实时PCR技术分别测定FN1、NME2、TIMP3基因表达水平。结果AMSCs组FN1、NME2、TIMP3基因表达水平均高于FMSCs、F6、SGC组(P<0.001);F6细胞以及白血病细胞中未能检出TIMP3基因的表达。结论FN1、NME2、TIMP3基因的下调以及TIMP3基因的缺如,可能是FMSCs突变为F6肿瘤细胞的分子特征之一;TIMP3基因的下调可能是肿瘤细胞的一种分子标志。

Objective To investigate the differential expression of genes encoding fibronectin 1 （FNI）, non-metastatic cell proteins 2 （NME2） and tissue inhibitor of metalloproteinase 3 （TIMP3） in fetal bone marrow mesenchymal stem cells （ FMSCs）, adult bone marrow mesenchymal stem cells （AMSCs）, F6, SGC and leukemia cells and study the molecular biological characters of tumor and leukemia cells. Methods RT-PCR was used to clone the cDNA of total RNA extracted from FMSCs, AMSCs, F6, SGC and leukemia cells respectively and then the expression levels of gene FN1, NME2 and TIMP3 were detected by PCR and real-time PCR. Results The expression levels of gene FN1 ,NME2 and TIMP3 in AMSCs were higher than those in FMSCs,F6,SGC （P 〈0. 001 ） ,but gene TIMP3 could not be detected in cDNA of F6 and leukemia cells. Conclusions The down-regulation of gene FN1, NME2 and TIMP3 or the deficiency of gene TIMP3 may be one of the molecular characteristics of the mutation of F6 from FMSCs, and the down-regulation of gene TIMP3 is probably one of the common markers of tumor cells.