Rui-lan Gao1, Xiao-jie Lin, Xiao-hong Chen. 1. Institute of Hematology, The First Affiliated Hospital, Zhejiang University of Chinese Medicine, Hangzhou. zphtcmcn@mail.hz.zj.cn
Abstract
OBJECTIVE: To explore the relationship of up-regulated genes with cell proliferation and differentiation 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 [alpha-33 ]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, including 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 family. 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 profile.
OBJECTIVE: To explore the relationship of up-regulated genes with cell proliferation and differentiation 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 [alpha-33 ]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, including 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 ratsarcoma (RAS) homology gene family. 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 profile.