目的:通过研究恶性胶质瘤细胞中死亡诱导信号复合体(DISC)与肿瘤坏死因子相关凋亡诱导配体(TRAIL)诱导凋亡的关系,初步探讨TRAIL诱导凋亡抵抗的机制。方法:分离恶性胶质瘤组织,获得和培养原代胶质瘤细胞,用100μg.L-1 TRAIL作用后采用...目的:通过研究恶性胶质瘤细胞中死亡诱导信号复合体(DISC)与肿瘤坏死因子相关凋亡诱导配体(TRAIL)诱导凋亡的关系,初步探讨TRAIL诱导凋亡抵抗的机制。方法:分离恶性胶质瘤组织,获得和培养原代胶质瘤细胞,用100μg.L-1 TRAIL作用后采用酸性磷酸酶法检测细胞凋亡水平;Western blotting法检测细胞表达死亡诱导信号复合体的水平。结果:获得3株(GC417、GC321、GC125)原代恶性胶质瘤细胞;3株细胞对TRAIL诱导凋亡的敏感程度不同[GC321(0.12±0.01 vs 0.51±0.02)和GC125(0.22±0.01 vs 0.36±0.01)],对TRAIL诱导凋亡作用敏感,与对照组比较差异有统计学意义(P<0.01);而GC417(0.24±0.01 vs 0.23±0.02)对TRAIL诱导凋亡不敏感。Western blotting法检测结果显示,死亡诱导信号复合体表达不同,GC321和GC125表达增高,GC417表达减少。结论:不同来源的原代培养恶性胶质瘤细胞对TRAIL诱导凋亡的反应不同,死亡诱导信号复合体表达也不同,死亡诱导信号复合体表达的减少可能与凋亡抵抗的发生密切相关。展开更多
Recent experiments showed that the apoptosis of tumor cells was highly related to the expression of tumor necrosis factor related apoptosis inducing factor (TRAIL) which was first cloned from expressed sequence tag da...Recent experiments showed that the apoptosis of tumor cells was highly related to the expression of tumor necrosis factor related apoptosis inducing factor (TRAIL) which was first cloned from expressed sequence tag database (EST). At the same time, it played a role in the lymphocytes differentiation and proliferation, but many of its characteristics and mechanism remain unknown up to now. To investigate its biological functions and related mechanisms, the coding region of human soluble TRAIL gene from LPS activated healthy human peripheral lymphocytes (PBL) was cloned using RT PCR and constructed a 6 His tag vector pProEX HTb hTRAIL. By DNA sequencing, the cloned hTRAIL was identical to that reported.Upon IPTG inducing, the soluble hTRAIL was highly expressed in E. coli K802 up to 30% of bacterial protein. The expressed 6 His hTRAIL protein could induce death of L929 cells as the same as TNF α. The recombinant TRAIL was obtained which paved the way for further studies on its biological characteristics and related mechanisms.展开更多
文摘目的:通过研究恶性胶质瘤细胞中死亡诱导信号复合体(DISC)与肿瘤坏死因子相关凋亡诱导配体(TRAIL)诱导凋亡的关系,初步探讨TRAIL诱导凋亡抵抗的机制。方法:分离恶性胶质瘤组织,获得和培养原代胶质瘤细胞,用100μg.L-1 TRAIL作用后采用酸性磷酸酶法检测细胞凋亡水平;Western blotting法检测细胞表达死亡诱导信号复合体的水平。结果:获得3株(GC417、GC321、GC125)原代恶性胶质瘤细胞;3株细胞对TRAIL诱导凋亡的敏感程度不同[GC321(0.12±0.01 vs 0.51±0.02)和GC125(0.22±0.01 vs 0.36±0.01)],对TRAIL诱导凋亡作用敏感,与对照组比较差异有统计学意义(P<0.01);而GC417(0.24±0.01 vs 0.23±0.02)对TRAIL诱导凋亡不敏感。Western blotting法检测结果显示,死亡诱导信号复合体表达不同,GC321和GC125表达增高,GC417表达减少。结论:不同来源的原代培养恶性胶质瘤细胞对TRAIL诱导凋亡的反应不同,死亡诱导信号复合体表达也不同,死亡诱导信号复合体表达的减少可能与凋亡抵抗的发生密切相关。
文摘Recent experiments showed that the apoptosis of tumor cells was highly related to the expression of tumor necrosis factor related apoptosis inducing factor (TRAIL) which was first cloned from expressed sequence tag database (EST). At the same time, it played a role in the lymphocytes differentiation and proliferation, but many of its characteristics and mechanism remain unknown up to now. To investigate its biological functions and related mechanisms, the coding region of human soluble TRAIL gene from LPS activated healthy human peripheral lymphocytes (PBL) was cloned using RT PCR and constructed a 6 His tag vector pProEX HTb hTRAIL. By DNA sequencing, the cloned hTRAIL was identical to that reported.Upon IPTG inducing, the soluble hTRAIL was highly expressed in E. coli K802 up to 30% of bacterial protein. The expressed 6 His hTRAIL protein could induce death of L929 cells as the same as TNF α. The recombinant TRAIL was obtained which paved the way for further studies on its biological characteristics and related mechanisms.
