肿瘤研究与临床
腫瘤研究與臨床
종류연구여림상
CANCER RESEARCH AND CLINIC
2009年
10期
649-653
,共5页
韩崇旭%许文荣%孙艳%张锡然
韓崇旭%許文榮%孫豔%張錫然
한숭욱%허문영%손염%장석연
骨髓%间质干细胞%肿瘤细胞%培养的%基因%Nucleostemin
骨髓%間質榦細胞%腫瘤細胞%培養的%基因%Nucleostemin
골수%간질간세포%종류세포%배양적%기인%Nucleostemin
Bone marrow%Mesenchymal stem cells%Tumor cells%cultured%Cells,Nucleostemin
目的 研究骨髓间充质干细胞(MSC)诱导肿瘤发生的机制.方法 用荧光差异显示技术(FDD)寻找差异基因;PCR、免疫组织化学、Western blotting加以验证;实时荧光定量反转录聚合酶链反应(Real time RT-PCR)检测诱导致瘤细胞在裸鼠体内致瘤后致瘤组织的基因表达水平.结果 FDD结果显示Nucleostemin(NS)基因高表达,与PCR、Western blotting显示结果一致.Real time RT-PCR表明,NS基因在MSC、F6及3组F6致瘤组织细胞之间的表达水平差异有统计学意义(F=160,P<0.05).F6组NS基因表达水平为(0.0372±0.0019),MSC基因表达水平为(0.0021±0.0002),增高18倍(P<0.05);裸鼠皮下注射F6细胞后,第4、6、7周分离致瘤组织细胞内NS基因表达水平分别为(0.0504±0.0083)、(0.0995±0.0026)和(0.0614±0.0036),呈上升趋势(与MSC对比,P值均<0.05).Western blotting及免疫组织化学染色证明F6细胞NS蛋白表达明显增高.结论 NS基因与肿瘤细胞形成有关.
目的 研究骨髓間充質榦細胞(MSC)誘導腫瘤髮生的機製.方法 用熒光差異顯示技術(FDD)尋找差異基因;PCR、免疫組織化學、Western blotting加以驗證;實時熒光定量反轉錄聚閤酶鏈反應(Real time RT-PCR)檢測誘導緻瘤細胞在裸鼠體內緻瘤後緻瘤組織的基因錶達水平.結果 FDD結果顯示Nucleostemin(NS)基因高錶達,與PCR、Western blotting顯示結果一緻.Real time RT-PCR錶明,NS基因在MSC、F6及3組F6緻瘤組織細胞之間的錶達水平差異有統計學意義(F=160,P<0.05).F6組NS基因錶達水平為(0.0372±0.0019),MSC基因錶達水平為(0.0021±0.0002),增高18倍(P<0.05);裸鼠皮下註射F6細胞後,第4、6、7週分離緻瘤組織細胞內NS基因錶達水平分彆為(0.0504±0.0083)、(0.0995±0.0026)和(0.0614±0.0036),呈上升趨勢(與MSC對比,P值均<0.05).Western blotting及免疫組織化學染色證明F6細胞NS蛋白錶達明顯增高.結論 NS基因與腫瘤細胞形成有關.
목적 연구골수간충질간세포(MSC)유도종류발생적궤제.방법 용형광차이현시기술(FDD)심조차이기인;PCR、면역조직화학、Western blotting가이험증;실시형광정량반전록취합매련반응(Real time RT-PCR)검측유도치류세포재라서체내치류후치류조직적기인표체수평.결과 FDD결과현시Nucleostemin(NS)기인고표체,여PCR、Western blotting현시결과일치.Real time RT-PCR표명,NS기인재MSC、F6급3조F6치류조직세포지간적표체수평차이유통계학의의(F=160,P<0.05).F6조NS기인표체수평위(0.0372±0.0019),MSC기인표체수평위(0.0021±0.0002),증고18배(P<0.05);라서피하주사F6세포후,제4、6、7주분리치류조직세포내NS기인표체수평분별위(0.0504±0.0083)、(0.0995±0.0026)화(0.0614±0.0036),정상승추세(여MSC대비,P치균<0.05).Western blotting급면역조직화학염색증명F6세포NS단백표체명현증고.결론 NS기인여종류세포형성유관.
Objective To study the tumorigenesis mechanism in bone marrow mesenchyme stem cells (MSC). Methods The bone marrow MSC could be induced into turnout (F6 cells) in vitro. The difference between gene expression of F6 cells and MSC was distinguished by fluorescent differential display (FDD). Verification of the result was detected by Real time RT-PCR and Western blotting, and immunocytochemistry. Results FDD analysis confirmed that Nucleostemin (NS) was positively up-regulated in F6 cells compared with MSC. Similar results were obtained by PCR and Western blotting. The NS gene expression levels in MSC, F6, 176-4, F6-6 and F6-7 were significantly different(F =160, P <0.05). The NS gene expression level in F6 (0.0372±0.0019) was 18 folds higher than those of MSC(0.0021±0.0002,P <0.05). Expression levels in F6-4, F6-6 and F6-7 tissue were 0.0504±0.0083, 0.0995±0.0026 and 0.0614±0.0036, and were significantly higher than that in MSC(P <0.05). The expression of NS increased significantly with the accreting volume of turnour, and high-level protein expression of NS was confirmed by Western blotting and immunocytochemistry. Conclusion The expression level of NS might be one of the factors playing important roles during turnour genesis, especially in MSC mutation.
