摘要
目的分离鉴定重组可溶性肿瘤坏死因子相关细胞凋亡诱导配体(rsTRAIL)在诱导JurkatT淋巴细胞白血病细胞凋亡过程中差异表达的基因。方法采用抑制性差减杂交(suppressionsubtractivehybridization,SSH)及PCR技术筛选差异表达的基因。采用狭缝杂交和Northern印迹技术鉴定差异基因片段;以DNA自动序列分析仪测定所获cDNA核苷酸顺序。结果获得了6个差异表达的基因片段,4个在TRAIL诱导的白血病细胞凋亡过程中被抑制,2个被激活;其中,A14、X1、D1、A23和C5等5个基因片段为新基因,其GeneBank登记号分别为AW731601、AW731602、AW731603、AW731604和BE239235。Northern印迹显示,基因D1在Jurkat和MCF-7中的表达明显高于其它肿瘤细胞,提示该基因可能具有肿瘤特异性。结论TRAIL诱导的细胞凋亡过程,涉及多种基因表达的改变。
or Objective To identify the genes differentially expressed in leukemia cell apoptosis induced by recombinant soluble tumor necrosis factor-related apoptosis inducing ligand(rsTRAIL).Methods Suppression subtractive hybridization(SSH)and polymerase chain reaction(PCR)were used for the cloning and identification of the genes differentially expressed in the apoptotic Jurkat cells induced by TRAIL.Slot blot and Northern blot were used for the expression pattern analysis of the genes.Automatic DNA sequencing was used for DNA sequence analysis.Results Six cDNA fragments differentially expressed in the Jurkat leukemia cells treated with TRAIL were found,in which four were inhibited and two were activated during the Jurkat cell apoptosis treated with TRAIL.Among which the five genes of A14,X1,D1,A23and C5were found at the first time by DNA sequencing and GeneBank database searching.So that they were registered in GeneBank as AW731601,AW731602,AW731603,AW731604and BE239235,respectively.It was found that the gene D1was expressed higher in Jurkat leukemia cells and MCF-7breast cancer cells than that in K562leukemia,825gastric cancer and7721liver cancer cells.Conclusions Five novel cDNA fragments were found,and among which D1might be a tumor specific gene.
出处
《中国医学科学院学报》
CAS
CSCD
北大核心
2002年第3期238-241,共4页
Acta Academiae Medicinae Sinicae
基金
北京市自然科学基金(7992032)
国家重点基础性研究973项目基金(G1990534)资助
