Ohtsu H, Kuramasu A, Suzuki S, Igarashi K, Ohuchi Y, Sato M, Tanaka S, Nakagawa S, Shirato K, Yamamoto M, Ichikawa A, Watanabe T
Department of Pharmacology I, Tohoku University School of Medicine, Sendai 980-77, Japan.
J Biol Chem. 1996 Nov 8;271(45):28439-44. doi: 10.1074/jbc.271.45.28439.
Phenotype of P815 mouse mast cells changes markedly during culture in the peritoneal cavity of syngenic BDF1 mice. The cells, cultured for 1 week in the peritoneal cavity of syngenic BDF1 mice, proliferate and express high levels of L-histidine decarboxylase (HDC) and mouse mast cell protease (MMCP)-6 mRNAs, indicating the ability of P815 cells to differentiate toward mature connective tissue mast cells. Peritoneal fluid aspirated from P815-inoculated BDF1 mouse and added to cultured P815 cells in vitro was also found to induce HDC mRNA expression, suggesting that at least some of the humoral factors in the peritoneal fluid induce HDC mRNA transcription. Among the erythroid transcription factors, P815 cells expressed GATA-2 but not GATA-1 mRNA before and after the intraperitoneal incubation. In contrast, the expression of NF-E2 subunit p45 disappeared, while expression of subunit mafK was markedly reduced after incubation. Cotransfection assays using HDC-luciferase reporter and p45 and/or mafK expression constructs showed that NF-E2 affects the transactivation of HDC gene. These results suggest that NF-E2 is also an important transcription factor in mast cell differentiation.
P815小鼠肥大细胞在同基因BDF1小鼠腹腔内培养期间表型发生显著变化。在同基因BDF1小鼠腹腔内培养1周的细胞会增殖，并高水平表达L-组氨酸脱羧酶（HDC）和小鼠肥大细胞蛋白酶（MMCP）-6 mRNA，这表明P815细胞具有向成熟结缔组织肥大细胞分化的能力。从接种P815的BDF1小鼠腹腔中吸出的腹腔液，并添加到体外培养的P815细胞中，也发现可诱导HDC mRNA表达，这表明腹腔液中至少某些体液因子可诱导HDC mRNA转录。在红系转录因子中，P815细胞在腹腔内孵育前后均表达GATA-2，但不表达GATA-1 mRNA。相反，孵育后NF-E2亚基p45的表达消失，而亚基mafK的表达显著降低。使用HDC-荧光素酶报告基因以及p45和/或mafK表达构建体进行的共转染试验表明，NF-E2影响HDC基因的反式激活。这些结果表明，NF-E2也是肥大细胞分化中的一种重要转录因子。
