MTEC1细胞系分泌的化学趋化因子的基因克隆,鉴定及功能研究

A novel gene-mouse estrogen enhanced transcript (mEET) containing two typical estrogen responsive elements (ERE) was cloned from MTEC1 cells, a mouse thymus epithelial cell line that produces constitutively many chemokines. Addition of 17 b-estradiol (E2) to the MTEC1 cell cultures enhanced mEET mRNA expression and, meanwhile, significantly inhibited monocyte chemoattractant protein-1 (MCP-1) and SDF-1??production.To analyse the functional links between the expression of mEET and of MCP-1, we transfected MTEC1 cells with ERE-deleted antisense or a sense mEET cDNA construct and activated the transfected mEET cDNA in stable MTEC1 transfectants with doxycycline. Doxycycline-induced activation of the mEET gene profoundly inhibited MCP-1 and SDF-1??expression at both mRNA and protein levels and alleviated its chemotactic activity. Conversely, inactivation of the mEET gene substantially augmented MCP-1 and SDF-1??expression. Activation of the mEET gene markedly attenuated activity of nuclear NF-kB. In summary, we have first demonstrated that estrogen-imposed inhibition of MCP-1 and SDF-1??expression occurs through the activation of the mEET gene, its product suppresses nuclear NF-kB and negatively regulates MCP-1 and SDF-1??gene activation.

研究ＭＴＥＣ１细胞分泌的化学趋化因子及其类型，用ＲＴ－ＰＣＲ及保守区简并引物等分子生物学方法，有可能确定新型因子，并进一步基因克隆和表达。研究趋化因子在骨髓细胞向胸腺及胸腺细胞由皮质向髓质定向迁移中的作用，及其在Ｔ细胞发育中对Ｔ细胞表型和功能成熟的作用和意义。生物功能的研究有利于阐明趋化因子在胸腺细胞发育过程中的作用。

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