Jikai J, Ryo R, Yasunaga M, Sugano W, Yamaguchi N, Hirano T
Department of Laboratory Medicine, Kobe University School of Medicine.
Rinsho Byori. 1992 Aug;40(8):841-7.
The usefulness of in situ hybridization for detecting mRNA encoding various kinds of proteins in megakaryocytic cell lines (K562, HEL and CMK) was examined. Using in situ hybridization, glycoprotein (GP) IIIa mRNA was found to be present in K562, HEL and CMK cells. Both GPIb and GPIIb mRNA were also present in HEL and CMK cells, while only HEL cells expressed platelet factor 4 mRNA. These findings were identical to those obtained with Northern blotting. It should be emphasized that in situ hybridization was useful to clearly define each cell expressing platelet specific protein mRNA. The expression of interleukin-6 (IL-6) and IL-6 receptor mRNA in mononuclear cells prepared from bone marrow aspirates was examined. In situ hybridization study demonstrated the presence of IL-6 receptor mRNA in the recognized megakaryocytes only, while IL-6 mRNA was found to be present in the megakaryocytes and a few mononuclear cells. These findings suggest that differentiation and proliferation of normal megakaryocytes might be controlled by an IL-6 autocrine loop, and detection of IL-6 receptor mRNA might be useful to identify megakaryocytes.
研究了原位杂交技术在检测巨核细胞系（K562、HEL和CMK）中编码各种蛋白质的mRNA方面的实用性。通过原位杂交发现，糖蛋白（GP）IIIa mRNA存在于K562、HEL和CMK细胞中。GPIb和GPIIb mRNA也存在于HEL和CMK细胞中，而只有HEL细胞表达血小板因子4 mRNA。这些结果与通过Northern印迹法获得的结果一致。应当强调的是，原位杂交有助于清晰地界定每个表达血小板特异性蛋白质mRNA的细胞。检测了从骨髓抽吸物中制备的单核细胞中白细胞介素-6（IL-6）和IL-6受体mRNA的表达情况。原位杂交研究表明，IL-6受体mRNA仅存在于已识别的巨核细胞中，而IL-6 mRNA存在于巨核细胞和少数单核细胞中。这些结果提示，正常巨核细胞的分化和增殖可能受IL-6自分泌环调控，检测IL-6受体mRNA可能有助于识别巨核细胞。
