[Expression and detection of platelet specific genes in human megakaryocytic cells].
The expression and biological function of interleukin-6(IL-6), and its receptor mRNA, were studied in a human megakaryocytic cell line (CMK). IL-6 possessed stimulatory effects on the DNA synthesis as well as colony formation of CMK cells. The IL-6 receptor mRNA could be detected by the method of reverse transcriptase polymerase chain reaction (RT-PCR) but not Northern blotting. On the contrary, IL-6 mRNA was detected by the method of RT-PCR, and its expression induced by the addition of 12-O-tetradecanoylphorbol-13-acetate (TPA) could be clearly shown by Northern blotting. These findings indicate that IL-6 and its receptor mRNA should be analyzed by both methods, and the growth and differentiation of CMK cells may be controlled by an IL-6 autocrine loop. Next, the expression and biological role of low molecular GTP-binding proteins (smg p21A and -B) mRNAs were examined in CMK cells. Both the smg p21A and -B mRNAs were detected in CMK cells using Northern blotting, and their levels were markedly elevated by TPA treatment. The mRNA level of glycoprotein IIb, a typical marker of the megakaryocytes, was increased by TPA, but the time course of the increase in the smg p21 mRNA levels was more rapid that that in the GPIIb mRNA level. These findings suggest that smg p21s play an important role during the TPA-induced differentiation of CMK cells.
Adachi
M
M
Department of Laboratory Medicine, Kobe University School of Medicine.
Yasunaga
M
M
Nakayama
K
K
Jikai
J
J
Yamaguchi
N
N
Ryo
R
R
D001792:Blood Platelets / Q000378:metabolism*; D002454:Cell Differentiation; D019204:GTP-Binding Proteins / Q000235:genetics; D005796:Genes; D006801:Humans; D015850:Interleukin-6 / Q000235:genetics; D008533:Megakaryocytes / Q000166:cytology / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000032:analysis; D011971:Receptors, Immunologic / Q000235:genetics; D019947:Receptors, Interleukin-6; D020668:rap GTP-Binding Proteins
D015850:Interleukin-6; D012333:RNA, Messenger; D011971:Receptors, Immunologic; D019947:Receptors, Interleukin-6; D019204:GTP-Binding Proteins; D020668:rap GTP-Binding Proteins