论文部分内容阅读
目的 :探索 p16基因在脑胶质瘤发生发展过程中的作用。方法 :分别采用脂质体、磷酸钙 +DMSO休克转染的方法 ,观察外源野生型 p16基因短暂转染与长期稳定转染对人胶质瘤细胞系U2 5 1、SWO的作用 ,并筛选阳性克隆。同时以空载体质粒PCDNA3为对照。免疫组化、Northern杂交检测p16基因表达 ,对转染后细胞生长、细胞周期及细胞凋亡的变化进行分析。结果 :p16基因短暂转染有明显的凋亡诱导作用 ;而p16基因长期稳定转染无明显的凋亡诱导作用 ,但外源 p16基因的高水平表达显著抑制了胶质瘤U2 5 1、SWO细胞的生长 ,克隆形成率减少 ,肿瘤细胞发生了G1期阻滞。结论 :外源野生型p16基因可抑制胶质瘤细胞恶性增殖并诱导细胞凋亡。肿瘤细胞的异质性特点与 p16基因转染后的“自然选择”作用可能是p16基因长期稳定转染无明显凋亡诱导作用的主要机制
Objective: To explore the role of p16 gene in the development of glioma. METHODS: The effects of transient transfection of exogenous wild-type p16 gene and long-term stable transfection on human glioma cell lines U251 and SWO were observed by liposome, calcium phosphate and DMSO shock transfection, respectively. Positive clones. At the same time, the empty vector plasmid PCDNA3 was used as a control. The expression of p16 gene was detected by immunohistochemistry and Northern blot analysis. Changes in cell growth, cell cycle, and apoptosis after transfection were analyzed. RESULTS: The transient transfection of p16 gene had obvious apoptosis-inducing effect. However, the long-term stable transfection of p16 gene had no obvious apoptosis-inducing effect, but the high-level expression of exogenous p16 gene significantly inhibited glioma U251 1 and SWO. Cell growth, colony formation was reduced, and G1 arrest occurred in tumor cells. Conclusion : Exogenous wild-type p16 gene can inhibit the malignant proliferation of glioma cells and induce apoptosis. The heterogeneity of tumor cells and the “natural selection” effect of p16 gene transfection may be the main mechanism of long-term stable transfection of p16 gene without obvious apoptosis induction.