摘要
目的 探讨内源性TGF β1、TNF α在三氧化二砷 (As2 O3)诱导HL 6 0细胞凋亡过程中的作用。方法 建立As2 O3 诱导HL 6 0细胞凋亡模型 ,应用RT PCR、定量PCR、ELISA、缺口末端标记(TUNEL)法、片段化DNA分析等方法研究As2 O3 诱导HL 6 0细胞凋亡过程中内源性TGF β1、TNF α及其受体基因以及TGF β1蛋白表达的变化 ;进而研究TGF β1、TNF α反义磷酸硫代脱氧寡核苷酸 (PSODN)对细胞凋亡的干预作用。结果 ①As2 O3 诱导HL 6 0细胞凋亡过程中伴有TGF β1、TNF α表达上调 (处理前后mRNA拷贝数分别为 135 46± 12 4,4972 16± 187和 2 32 73± 2 2 9,6 742 17± 189) ,bcl 2表达下调 (处理前、后分别为 10 42 4± 2 74和 336 1± 89) ,细胞培养 2 4h上清中TGF β1蛋白水平明显提高 ,上升为对照组的 2 .0倍。②TGF β1、TNF α反义PSODN能够明显抑制As2 O3 诱导细胞凋亡的发生 ,并使细胞bcl 2mRNA及蛋白表达恢复到处理前的水平。结论 内源性TGF β1、TNF α在As2 O3 诱导HL 6 0细胞凋亡中起重要作用 ,两者均可通过下调bcl 2表达促进细胞凋亡。
Objective To study the effect of endogenous TGF-β 1 and TNF-α on As 2O 3 inducing apoptosis of HL-60 cells. Methods The expressions of endogenous TGF-β 1 and TNF-α in apoptotic HL-60 cells induced by As 2O 3 were assayed by RT-PCR, quantitative RT-PCR, ELISA, DNA fragmentation and TUNEL. The effect of TGF-β 1 and TNF-α antisense phosphorothioate oligodeoxynucleotides (PSODNs) on As 2O 3 inducing apoptotic HL-60 cells was further studied. Results ① Expressions of endogenous TGF-β 1 and TNF-α were significantly up-regulated in As 2O 3 inducing apoptotic HL-60 cells (from 13?546±124 and 497?216±187 before treatment to 23?273±229 and 674?217±189 after treatment, respectively), accompanied with down-regulated bcl-2 mRNA expression(from 10?424±274 before treatment to 3?361±89 after treatment). ②TGF-β 1 and TNF-α antisense PSODNs could rescue As 2O 3 induced apoptosis of HL-60 cells, with a restoration of bcl-2 gene expression. Conclusions Endogenous TGF-β 1 and TNF-α played an important role in As 2O 3 inducing HL-60 cells apoptosis through down-regulation of bcl-2 expression.
出处
《中华血液学杂志》
CAS
CSCD
北大核心
2003年第5期231-234,共4页
Chinese Journal of Hematology
基金
国家教委优秀年轻教师基金资助项目 ( 95 - 5 0 3)
福建省教委科研基金资助项目 (K95 0 33)
国家"百千万人才工程"专项基金资助项目
