不同年龄段人前列腺外周带基质细胞表型特征的研究

Characterization of stromal cell cultures from the prostatic periheral zone of men at different ages

目的:探讨体外培养的不同年龄段人前列腺外周带基质细胞的表型特征和生长特性差异,分析前列腺基质细胞微环境对老年男性前列腺癌促发的可能原因。方法:原代培养不同年龄段前列腺外周带基质细胞,利用免疫细胞化学技术分析细胞表型特征、透射电镜观察细胞超微结构、琥珀酸脱氢酶法观察细胞体外增殖以及流式细胞术检测细胞凋亡情况。结果:不同年龄段前列腺基质细胞均表达成纤维细胞标记物prolyl-4-hydroxy-lase,而平滑肌肌动蛋白(α-SMA)、肌间线蛋白(desmin)的表达随年龄逐渐增加,即α-SMA的阳性表达:年轻组vs老年组=(2.56±1.81)%vs(38.89±11.22)%,(P<0.01);而desmin阳性表达:年轻组vs老年组=(0.89±0.93)%vs(14.89±5.97)%,(P<0.01)。并且α-SMA和/或desmin阳性细胞形态相对宽大、扁平、多形性。超微结构观察发现,前列腺基质细胞内蛋白合成有关的细胞器随着年龄的增加不断增多,主要为粗面内质网和高尔基复合体。老年组前列腺基质细胞的增殖率低于年轻组(P<0.01);而两组基质细胞凋亡率均在1%～3%之间,组间差异无统计学意义。结论:前列腺外周带基质细胞中具有活性的肌成纤维细胞随年龄不断增加可能是老年男性前列腺癌高发和恶性进展的原因之一。

Objective：To characterize age-related cellular phenotype alterations and growth rates of human prostatic stromal cell cultures from the normal prostatic peripheral zone of young donors（PZ-young） and old donors（PZ-old）.Methods：We isolated stromal cells from 10 donors of different ages,assessed the cellular phenotypes by immunocytostaining for prolyl-4-hydroxylase,alpha-smooth muscle actin（α-SMA） and desmin,and analysed the ulturastructure by transmission electron microscopy（TEM）.The proliferation and apoptosis of the cells were determined by Cell Counting Kit-8 assay and flowcytometry,respectively.Results：All the stromal cells were positive for prolyl-4-hydroxylase regardless of the donors＇ age,while α-SMA and desmin positive cells increased with their age.The positive expressions of α-SMA and desmin were（2.56±1.81） % and（0.89±0.93） % in PZ-young,and（38.89±11.22） % and（14.89±5.97） % in PZ-old（P0.01）.The α-SMA-and/or desmin-positive stromal cells were morphologically large,flat and polygonal.Ultrastructural analysis showed that the cell cultures from PZ-old were richer in rough endoplasmic reticulum and golgi complexes.The stromal cells of PZ-old had a lower growth rate than that of PZ-young（P0.01）,but there was no significant difference in the apoptosis rate between the two groups.Conclusion：Cellular phenotypes of human prostate stromal cell cultures change with the increase of age from predominantly typical fibroblasts to a mixture of fibroblasts and myofibroblasts,which might responsible for the high incidence of prostate cancer in elderly men.