Thrombopoietin and chemokine mRNA expression in patient post-chemotherapy and in vitro cytokine-treated marrow stromal cell layers.
CD34(+) cells and megakaryocyte progenitors were lower in marrow from patients after hematological recovery from the first cycle of 5-fluorouracil, leucovorin, adriamycin, cyclophosphamide (FLAC) chemotherapy plus PIXY321 (GM-CSF/interleukin 3; IL-3 hybrid) than in FLAC + GM-CSF or pre-FLAC marrows. Marrow stromal layers, an in vitro model of the marrow microenvironment, express a combination of stimulatory and inhibitory factors that modulate hematopoietic progenitor cell proliferation and differentiation. The TaqMan assay and quantitative reverse transcriptase-polymerase chain reaction were used to measure monocyte chemoattractant protein-1 (MCP-1), melanoma stimulatory growth activity, and monokine inducible by interferon-gamma (Mig) (inhibitory chemokines for primitive or megakaryocyte progenitors) mRNA levels in in vitro PIXY and GM-CSF-treated and patient post-FLAC marrow stromal layers. Chemokine mRNA was increased after in vitro GM-CSF and to a lesser extent after PIXY treatment. MCP-1 mRNA levels were fivefold higher in FLAC + PIXY than in FLAC + GM-CSF layers, and Mig mRNA was elevated in FLAC + GM-CSF layers. Thrombopoietin (TPO), insulin-like growth factor I (IGF-I), and IGF-II (stimulatory factors for primitive and megakaryocyte progenitors) mRNA were also measured. TPO mRNA levels were 30% lower in GM-CSF and PIXY-pretreated than in control layers with no decrease in IGF mRNA. TPO mRNA in stromal layers of patients who developed grade 3 thrombocytopenia (platelets < 20 x 10(9)/l) during the third cycle of FLAC was only 24% of levels in stromal layers of marrow from other post-FLAC patients. Results demonstrate that patient and in vitro treatment had modulatory effects on TPO and chemokine mRNA expression in marrow stromal layers.
Schwartz
G N
GN
Department of Experimental Transplantation and Immunology, Medicine Branch and Surgery Branch, National Cancer Institute, Bethesda, Maryland, USA. gnschwar@cpcug.org
Kammula
U
U
Warren
M K
MK
Park
M K
MK
Yan
X Y
XY
Marincola
F M
FM
Gress
R E
RE
D000971:Antineoplastic Combined Chemotherapy Protocols / Q000627:therapeutic use*; D001854:Bone Marrow Cells / Q000166:cytology / Q000473:pathology / Q000502:physiology*; D001943:Breast Neoplasms / Q000188:drug therapy* / Q000473:pathology*; D002478:Cells, Cultured; D018932:Chemokine CCL2 / Q000235:genetics; D018925:Chemokines / Q000235:genetics*; D003520:Cyclophosphamide / Q000008:administration & dosage; D016207:Cytokines / Q000494:pharmacology*; D004317:Doxorubicin / Q000008:administration & dosage; D005260:Female; D005472:Fluorouracil / Q000008:administration & dosage; D005786:Gene Expression Regulation / Q000187:drug effects / Q000276:immunology; D016179:Granulocyte Colony-Stimulating Factor / Q000494:pharmacology; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000494:pharmacology / Q000627:therapeutic use*; D006801:Humans; D007334:Insulin-Like Growth Factor I / Q000235:genetics; D007335:Insulin-Like Growth Factor II / Q000235:genetics; D007377:Interleukin-3 / Q000494:pharmacology / Q000627:therapeutic use*; D002955:Leucovorin / Q000008:administration & dosage; D012333:RNA, Messenger / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000627:therapeutic use; D011994:Recombinant Proteins; D017154:Stromal Cells / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D013926:Thrombopoietin / Q000235:genetics*; D014158:Transcription, Genetic / Q000187:drug effects / Q000276:immunology
D018932:Chemokine CCL2; D018925:Chemokines; D016207:Cytokines; D007377:Interleukin-3; C078524:PIXY321 fusion protein, recombinant; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D016179:Granulocyte Colony-Stimulating Factor; D007334:Insulin-Like Growth Factor I; D007335:Insulin-Like Growth Factor II; D004317:Doxorubicin; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D003520:Cyclophosphamide; D013926:Thrombopoietin; D002955:Leucovorin; D005472:Fluorouracil