Harley C B, Menon C R, Rachubinski R A, Nieboer E
Department of Biochemistry, McMaster University, Hamilton, Ontario, Canada.
Biochem J. 1989 Sep 15;262(3):873-9. doi: 10.1042/bj2620873.
To help to evaluate the role of metallothionein (MT) in peripheral-blood leucocytes, we examined MT protein and mRNA levels in these cells before and after exposure to CdCl2 in culture. Protein was assayed by 109Cd2+ binding, and RNA by dot-blot hybridization. MT was induced in both lymphocytes and adherent monocytes about 10-fold with a 12 h exposure to 10 microM-CdCl2, but absolute levels were 3-fold higher in monocytes: 57 x 10(5) (+Cd2+) versus 6 x 10(5) (-Cd2+) molecules/cell for monocytes; 18 x 10(5) (+Cd2+) versus 2 x 10(5) (-Cd2+) for lymphocytes. Polymorphonuclear cells expressed relatively little MT (0.6 x 10(5) molecules/cell), and this did not change with phorbol ester stimulation or exposure to Cd2+, arguing against a direct protective role for MT in activated neutrophils. MT mRNA levels corresponded qualitatively to expression of protein in these cells. Our data provide quantitative comparisons of leucocyte MT expression and regulation in the human population. Variation in both basal and induced MT mRNA levels reflects environmental or experimental (intra-individual) and possibly genetic (inter-individual) differences.
为了评估金属硫蛋白(MT)在外周血白细胞中的作用,我们检测了培养的外周血白细胞在暴露于氯化镉(CdCl2)前后的MT蛋白和mRNA水平。通过109Cd2+结合法检测蛋白,通过斑点杂交法检测RNA。淋巴细胞和贴壁单核细胞在暴露于10 microM - CdCl2 12小时后,MT均被诱导增加约10倍,但单核细胞中的绝对水平高出3倍:单核细胞为57×10(5)(+Cd2+)分子/细胞,而(-Cd2+)时为6×10(5)分子/细胞;淋巴细胞为18×10(5)(+Cd2+)分子/细胞,而(-Cd2+)时为2×10(5)分子/细胞。多形核细胞表达的MT相对较少(0.6×10(5)分子/细胞),并且在佛波酯刺激或暴露于Cd2+后也没有变化,这表明MT在活化的中性粒细胞中没有直接的保护作用。MT mRNA水平在质量上与这些细胞中蛋白质的表达相对应。我们的数据提供了人群中白细胞MT表达和调节的定量比较。基础和诱导的MT mRNA水平的变化反映了环境或实验(个体内)以及可能的遗传(个体间)差异。