论文部分内容阅读
目的 :探讨 P16基因对人胶质瘤细胞的抑制作用 ,为胶质瘤致瘤机理的研究和基因治疗提供理论基础。方法 :运用分子克隆技术构建重组人 P16基因表达载体 (pc DNA3- P16 ) ,通过脂质体介导法将 P16 c DNA转染到存在 P16基因纯合缺失的人胶质瘤细胞系 SHG- 44。结果 :PCR证实含 P16 c DNA质粒 DNA已转入 SHG- 44细胞 ,Western blot显示有 P16蛋白表达。细胞生长曲线显示转染 P16基因的 SHG- 44细胞 (SHG- 44 - P16 )生长明显受到抑制 ,其抑制率达 84.2 7%。在软琼脂上克隆形成能力下降约 6倍。细胞周期分析表明 ,处在 G1期细胞由 30 .70 %提高到 6 6 .2 1%。结论 :野生型 P16基因导入 P16基因功能缺失的肿瘤细胞 ,能恢复其抑制肿瘤细胞生长的功能。
Objective: To investigate the inhibitory effect of P16 gene on human glioma cells and to provide a theoretical basis for the study of glioma tumorigenesis and gene therapy. METHODS: Recombinant human P16 gene expression vector (pcDNA3-P16) was constructed by molecular cloning technique. P16 c DNA was transfected into human glioma cell line SHG-44 with homozygous deletion of P16 gene by liposome-mediated method . Results: PCR confirmed that plasmid DNA containing P16 c DNA had been transfected into SHG-44 cells and Western blot showed P16 protein expression. Cell growth curve showed that the growth of SHG-44 cells (SHG-44 - P16) transfected with P16 gene was significantly inhibited and the inhibition rate was 84.27%. Clone formation on soft agar decreased about 6-fold. Cell cycle analysis showed that cells in G1 phase increased from 30.7% to 6.62%. Conclusion: The introduction of wild-type P16 gene into P16 gene-deficient tumor cells restored its function of inhibiting tumor cell growth.