Quantitative analysis of the two macrophage colony-stimulating factor mRNA expressed in a human stromal cell line by reverse transcription-polymerase chain reaction (RT-PCR).
We established a quantitative analysis system for 4.0 kb and 1.6 kb macrophage colony-stimulating factor (M-CSF) mRNA, using reverse transcription-polymerase chain reaction. Using this system, we performed quantitative analysis of the two mRNAs expressed in the human stromal cell line, KM102, in the resting condition and when stimulated by various concentrations of interferon-gamma (IFN-gamma). The expression of 1.6 kb M-CSF mRNA was more efficiently stimulated by IFN-gamma than that of 4.0 kb M-CSF mRNA. The alternative splicing of a single M-CSF gene has been shown to generate several M-CSF proteins with different localization; we believe that molecular analysis of the transcription products by this system is important to better understand the physiological significance of the different species of M-CSF derived from each mRNA.
Department of Internal Medicine, Jichi Medical School, Tochigi, Japan.
T
T
Ohtsuki
M
M
Ikeda
K
K
Hatake
H
H
Tomizuka
Y
Y
Hoshino
S
S
Suzu
K
K
Harigaya
K
K
Motoyoshi
Y
Y
Miura
D001483:Base Sequence; D002460:Cell Line; D006801:Humans; D015151:Immunoblotting; D007371:Interferon-gamma / Q000494:pharmacology; D016173:Macrophage Colony-Stimulating Factor / Q000235:genetics*; D008969:Molecular Sequence Data; D010316:Particle Size; D016133:Polymerase Chain Reaction / Q000379:methods; D012333:RNA, Messenger / Q000032:analysis*
D012333:RNA, Messenger; D016173:Macrophage Colony-Stimulating Factor; D007371:Interferon-gamma