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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 investigate the differences in phenotypic characteristics and growth characteristics of stromal cells in peripheral stroma of human prostate in different age groups, and to analyze the possible causes of prostatic stromal cell microenvironment on the promotion of prostate cancer in elderly men. Methods: Primary cultured stromal cells of peripheral stroma of prostate were cultured in different ages. Immunocytochemistry was used to analyze the phenotypic characteristics of the cells. Transmission electron microscopy was used to observe the ultrastructure of the cells. The proliferation of cells in vitro was observed by succinate dehydrogenase method and the cells were detected by flow cytometry Apoptosis. Results: Prolial stromal cells expressed prolyl-4-hydroxy-lase at different ages, while the expression of α-SMA and desmin increased with age, that is, α (2.56 ± 1.81)% vs (38.89 ± 11.22)%, respectively (P <0.01). The positive expression of desmin in the young group vs the elderly group was (0.89 ± 0.93)% vs (14.89 ± 5.97)%, (P <0.01). And α-SMA and / or desmin-positive cells were relatively broad, flat and pleomorphic. Ultrastructural observation revealed that the organelles involved in protein synthesis in prostate stromal cells increased with age, mainly including the rough endoplasmic reticulum and the Golgi complex. Proliferation of prostate stromal cells in the elderly group was lower than that in the young group (P <0.01). The apoptosis rate of stromal cells in both groups was between 1% and 3%, with no significant difference between the two groups. CONCLUSIONS: The increasing number of myofibroblasts in stromal cells in peripheral stroma of prostate may be one of the reasons for the high and malignant progression of prostate cancer in elderly men.