Stathmin基因在人成骨肉瘤细胞中的表达及意义

Significance of Stathmin Gene Overexpression in Osteosarcoma Cells

背景与目的:Stathmin作为细胞信号转导分子在细胞分化及恶性肿瘤发生、发展上发挥重要作用。本研究旨在探讨Stathmin基因在人成骨肉瘤细胞中的表达,并探讨阻断其表达对该肿瘤细胞的生物学行为的影响。方法:RT-PCR及原位杂交法检测2个人成骨肉瘤细胞系及45例人成骨肉瘤组织中Stathmin基因表达情况;以高表达Stathmin基因的人成骨肉瘤细胞系SOSP-9607为靶细胞、反义Stathmin(ASODN)为阻断剂,通过MTT实验观察ASODN对该细胞系的生长抑制作用,并用流式细胞仪分析其对细胞增殖周期的影响。结果:Stathmin基因在2个成骨肉瘤细胞系高表达,45例成骨肉瘤组织中24例呈阳性表达(阳性率为53.3%),10例正常组织中2例呈弱阳性表达,Stathmin在人成骨肉瘤组织的表达与正常组织中的表达有显著性差异(P<0.05)。ASODN明显抑制成骨肉瘤细胞的生长(P<0.05)。SOSP-9607在ASODN作用下,细胞分裂阻滞在分裂期的中期,并诱导细胞发生凋亡。结论:Stathmin基因在人成骨肉瘤中高表达,该基因有望成为成骨肉瘤生物治疗中的重要靶点。

BACKGROUND &OBJECTIVE: Stathmin, a signal transduction regulatory factor, plays a crucial role in cell division and malignant tumor development. This study was designed to analyze stathmin gene expression in the tissue of osteosarcoma, and to explore the growth inhibition by blocking stathmin expression with Stathmin antisense oligodeoxynucleotide (ASODN) on cultured human osteosarcoma cell line with stathmin gene overexpression. METHODS: Reverse transcriptase polymerase chain reaction (RT PCR) and in situ hybridization methods were used to determine the expression of stathmin in two human osteosarcoma cell lines and 45 osteosarcoma tissue specimens. Using stathmin gene overexpression in human osteosarcoma cell line SOSP 9607 as target cell and stathmin ASODN as gene expression blocking agent, cell growth inhibition of stathmin ASODN was determined by MTT method, and cell growth and mitotic character was analyzed by flow cytometry. RESULTS: Both osteosarcoma cell lines of SOSP 9607 and SOSP 9901 showed stathmin gene overexpression by RT PCR method and 24 of 45 osteosarcoma specimens were stathmin positive by in situ hybridization, while there were only 2 in 10 normal tissues with mild positive signal. There was significant difference in stathmin gene expression between human osteosarcoma tissues and normal tissues (P< 0.05). The growth of osteosarcoma cells were suppressed by ASODN (P< 0.05,P< 0.01). The cell cycle of SOSP 9607 was blocked in metaphase and cell apoptosis was found. CONCLUSION: Stathmin gene displays high level expression in osteosarcoma, which may become a new target for the bio treatment for osteosarcoma.