摘要
目的采用cDNA芯片技术分析三七总皂苷(PNS)诱导造血细胞的基因表达谱,探讨上调基因与细胞增殖、分化的相关性。方法选择增殖、分化相关重要的功能基因480个,制备cDNA膜芯片。人造血细胞巨核系CHRF-288、粒系HL-60和红系K562细胞经PNS处理后,分别提取mRNA,经逆转录后用〔α-33P〕dATP标记,与芯片的cDNA杂交。结果PNS诱导表达上调3倍以上的基因按功能分11类,包括甲基和乙酰化转移酶、诱导分化、抑制凋亡、转录调控蛋白、周期蛋白、信号传递介导蛋白和激酶、受体、DNA和RNA聚合酶,以及大鼠肉瘤(RAS)基因家族等。PNS诱导CHRF-288、HL-60和K562细胞后,mRNA表达水平上调3倍以上的基因分别为78条、89条和59条,占检测基因总数的16.3%、18.5%和12.3%。结论PNS诱导上调的基因均与细胞增殖和分化相关,与我们报道的血液病动物模型、造血干/祖细胞、基因转录调控和蛋白激酶等研究结果相符,为PNS的作用提供了基因表达谱方面的有力证据。
Objective To explore the relationship of up-regulated genes with ceil proliferation and differen- tiation by analyzing the hematopoietic cells gene expression profile induced by panax notoginosides （PNS） using cDNA microarray. Methods The cDNA membrane microarray with 480 target genes related to proliferation and differentiation of hematopoietic cells was prepared, and mRNA was extracted and purified from 3 lineages of human hematopoietic cell lines, including megakaryocytic CHRF-288, granulocytic HL-60 and erythrocytic K562 cells, respectively after they were treated with PNS. The hybridization with target genes on microarray membrane was performed using （a-^33p）dATP labeled cDNA from reversed mRNA. Results After treated by PNS, the genes up-regulated for more than 3 folds could be classified to 11 sorts according to their function, in- cluding the methyl-transferase, acetyl-transferase, differentiation initiated factor, anti-apoptosis, transcription regulation protein, cell cycle related protein, protein and kinase of signal pathway, receptors, DNA or RNA polymerase, protein phosphatase, transporter or trafficking protein and rat sarcoma （RAS） homology gene fami- ly. In three cell lines of CHRF-288, HL-60 and K562 treated by PNS, 78 （16.3%）, 89 （18.5%） and 59 （ 12.3 % ） pieces of genes respectively were up-regulated for more than 3 folds. Conclusion All the up-regulated genes induced by PNS in microarray analysis were related to hematopoietic cell proliferation and differentiation, the outcome is in accord with the results reported previously by the authors from the studies of mice model with hematopathy, hematopoietic stem/progenitor cells, gene transcription regulation and protein kinase of signal pathway, etc. It provides a powerful evidence for the PNS activity and its mechanism by gene expression pro- file.
出处
《中国中西医结合杂志》
CAS
CSCD
北大核心
2007年第11期999-1003,共5页
Chinese Journal of Integrated Traditional and Western Medicine
基金
国家自然科学基金资助项目(No.30070933)
关键词
三七总皂苷
基因芯片
造血干/祖细胞
增殖
分化
panax notoginosides
cDNA microarray
hematopoietic stem/progenitor cell
proliferation
differentiation
