[Differentiation of HL-60 cells directly cocultured with HFCL cells and alteration of their gene expression profile].
To study the molecular mechanism of the effect of fibroblastoid stromal cells (HFCL) from human bone marrow on the proliferation and differentiation in acute myeloid leukemia HL-60 cells, the cell cycle was analyzed by flow cytometry (FCM); the cell differentiation was determined by morphology NBT test and flow cytometric detection for expression of CD11b, CD14, CD13 and CD33; the genes differently expressed between HL-60 cells and HL-60 cells directly cocultured with HFCL were detected by using Affymetric oligo microarray technique. The changes of expression in some key genes were confirmed by using RT-PCR and Northern blot. The results showed that the percentage of G(1) phase cells in AML cells cocultured with HFCL cells was higher than that without HFCL cells, and the percentage of S phase cells was lower. The NBT positive cells and the expression of CD11b and CD14 increased. It was found that after direct contact of HL-60 cells with HFCL cells for 96 hours, the expression levels of 582 genes were up-regulated, 1 323 genes down-regulated. It is concluded that many genes may take part in the influence of HFCL cells on HL-60 cells, which may give important insights into the important molecules and pathways or cross-talk involved in the interaction between the AML cells and stromal cells.
Liang
Rong
R
Department of Hematology, Xijing Hospital, The Fourth Military Medical University, Xi'an 710033, China.
Huang
Gao-Sheng
GS
Chen
Xie-Qun
XQ
Bai
Qing-Xian
QX
Wang
Zhe
Z
Dong
Bao-Xia
BX
Wang
Wen-Qing
WQ
Zhang
Wei-Ping
WP
D002471:Cell Transformation, Neoplastic / Q000235:genetics*; D018920:Coculture Techniques; D005347:Fibroblasts / Q000166:cytology / Q000502:physiology*; D015870:Gene Expression; D020869:Gene Expression Profiling; D018922:HL-60 Cells; D006801:Humans; D017154:Stromal Cells / Q000166:cytology / Q000502:physiology*