结合珠蛋白在正常人表皮细胞及HaCaT细胞中的表达

Expression of Haptoglobin in Normal Human Epidermal Cells and HaCaT Cells

目的检测正常人表皮细胞及HaCaT细胞中结合珠蛋白(Hp)mRNA及蛋白的表达。方法用原位杂交及RT-PCR法检测正常人表皮细胞及HaCaT细胞中HpmRNA的表达,用免疫组化法检测正常人表皮中Hp的表达;用免疫组化法及蛋白质免疫印迹法检测HaCaT细胞中Hp的表达。结果在正常人表皮角质形成细胞(KC)及HaCaT细胞中HpmRNA表达阳性;正常人表皮朗格汉斯细胞(LC)中HpmRNA表达阴性;正常人表皮内可见Hp阳性的树突状细胞。用免疫组化法在每个HaCaT细胞胞质中未见明显Hp染色,但用蛋白质免疫印迹法在HaCaT细胞中检测到Hp蛋白。结论正常人KC及HaCaT细胞有合成Hp能力,正常人表皮LC无合成Hp的能力,在HaCaT细胞中有少量Hp表达。

Objective To investigate the expression of haptoglobin(Hp)mRNA and protein in nor-mal human epidermal cells and human keratinocyte cell line-HaCaT cells.Methods In situ hybridization and RT-PCR were used to detect the expression of Hp mRNA in normal human epidermal cells and HaCaT cells.Immunohistochemistry was used to detect the expression of Hp on normal human epidermal cells.Im-munohistochemistry and Western blot were used to detect Hp expression on HaCaT cells.Results There was Hp mRNA expression in normal human keratinocytes and HaCaT cells.There was no Hp mRNA expres-sion in normal human epidermal Langerhans cells.There were some Hp positive dendritic cells in normal human epidermis.There was no obvious Hp protein staining in the HaCaT cells by immunohistochemistry.There was Hp protein band from HaCaT cells by Western blot.Conclusions There is Hp mRNA expression in normal human keratinocytes and HaCaT cells which suggests that normal human keratinocytes and HaCaT cells have the ability to synthesize Hp protein.Normal human epidermal Langerhans cells have no ability to synthesize Hp protein.There is small amount of Hp protein in HaCaT cells.