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为了探讨人跨膜型和跨膜稳定型肿瘤坏死因子 - α(TNF- α)对脑恶性胶质瘤基因治疗的有效方法 ,研究应用了基因重组、细胞培养以及免疫组化等技术 ,通过真核表达载体 pc DNA3转染 NIH3T3细胞 ,并在体外观察了 TNF- α对脑胶质瘤生长的影响。结果显示 :与对照组相比 ,TM- TNF- α组和 TM- TNF- αm组中 GFAP和 S- 10 0免疫反应阳性肿瘤细胞的数量、胞体截面积、周长、平均光密度皆明显减少。统计学分析提示差异有显著性意义。这表明 TNF- α基因的表达可明显抑制胶质细胞的生长和分化。而且 ,还发现跨膜稳定型 TNF- α的杀瘤效果较跨膜型 TNF- α强。这为 TNF- α基因用于胶质瘤的基因治疗提供了一定的实验依据 ,提示大剂量的 TNF- α在临床治疗可能产生毒副作用。
In order to investigate the effective method of human transmembrane and transmembrane stable tumor necrosis factor-α (TNF-α) in gene therapy of malignant glioma, gene recombination, cell culture, Nuclear DNA expression vector pcDNA3 transfected NIH3T3 cells, and observed in vitro TNF-α on glioma growth. The results showed that compared with the control group, the number of GFAP and S-100 immunoreactive tumor cells, cell body cross-sectional area, perimeter and average optical density of TM-TNF-α group and TM-TNF-αm group were significantly decreased . Statistical analysis showed that the difference was significant. This indicates that the expression of TNF-α gene can significantly inhibit glial cell growth and differentiation. Moreover, transmembrane-stabilized TNF-alpha was also found to be more potent antitumor agent than transmembrane TNF- [alpha]. This provides some experimental evidence for the gene therapy of glioma with TNF-α gene, suggesting that high dose of TNF-α may have side effects in clinical treatment.