COMPARISON OF THE EXPRESSION OF
CYTOKINES MRNA IN FOUR MEGAKARYOCYTIC CELL LINES: MEG-01, DAMI, CHRF-288-11
AND M07E
Zhao D-C1,
Yang M2, Li K2, Chik K-W2, Yan M2,
Li C-K2, Shing M-M-K2, Fok T-F2, Yuen
P-M-N2
1
Department of Pediatrics, Maternal and Children Hospital of Guangdong
Province, Guangzhou, China
2
Department of Pediatrics, Prince of Wales Hospital, The Chinese University
of Hong Kong
Objective: Hematopoiesis is
regulated by a complex network of cytokines. Different studies performed in
different megakaryocytic cells have shown the cytokines pattern of
variability. Whether or not the megakaryocytic cell lines could produce
different pattern of cytokine in their different stages. The study was
tried to assay the repertoire of cytokines mRNA in these four cell lines:
MEG-01, DAMI, CHRF-288-11 and M-07E. Methods: Total RNA was extracted from
2-5×106 cells by RNeasyR Kits and quantitated with a spectrophotometer in
megakaryoblastic cell lines including MEG-01, DAMI, CHRF-288-11, and M-07E.
Cytokine mRNA expression for peripheral blood mononuclear cells (PBMCS) was
induced by ConA(10ng/ml) for 6 hours. Reverse transcription of mRNA was
performed to cDNA. cDNA was amplified by PCR method in volume of 50ul
containing 1ul of the cDNA, 2.5 ul of 10×PCR buffer, 0.5U of
Taq polymerase, 0.2mM/l dNTP and 15 pmol of each primer. PCR products were
run 2% agarose gel containing ethidium bromide and the amplified DNA could
be visualized by UV transillumination and photographed. Furthermore, the
products of RT-PCR were tested by southern blot analysis.