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目的探讨年轻和老年前列腺外周带基质细胞AR活性差异及对前列腺上皮细胞的增殖影响。方法不同年龄段前列腺基质细胞给予生理浓度10nM DHT刺激,荧光素酶报告基因检测AR转录活性差异,Elisa检测AR调控因子FGF-2,KGF,IGF-1分泌差异。CCK8检测与BPH-1及LNCaP共培养后BPH-1及LNCaP增殖情况。免疫组化,Q-PCR,Westernblot探宄不同年龄前列腺基质细胞ARA55表达,慢病毒干扰ARA55,给予10nM DHT刺激,Western-blot检测基质细胞AR表达差异,荧光素酶报告基因检测基质细胞AR转录活性差异,Elisa检测FGF-2,KGF,IGF-1分泌差异。CCK8检测阴性对照组(ARA55-SC)与干扰组(ARA55-sh)与BPH-1及LNCaP共培养后BPH-1及LNCaP增殖情况。结果基质细胞给予10nM DHT刺激,老年基质细胞AR转录活性、AR调控细胞因子FGF-2,KGF,IGF-1及对上皮细胞(BPH-1、LNCaP)促增殖高于年轻人(P<0.05)。老年基质细胞ARA55表达高于年轻人。干扰基质细胞ARA55后,不同年龄段基质细胞AR表达无差异,老年基质细胞ARA55-sh较ARA55-SC AR转录活性、AR调控细胞因子FGF-2,KGF,IGF—1分泌及对上皮细胞(BPH-1、LNCaP)促增殖降低(P0.05)。结论在10nM DHT刺激下,老年人基质细胞AR活性强于年轻人。老年基质细胞可能通过高表达ARA55增强AR转录活性和AR调控细胞因子分泌,促进BPH-1及LNCaP增殖。
Objective To investigate the difference of AR activity in stromal cells of peripheral prostatic stroma between young and old and the effect on the proliferation of prostatic epithelial cells. METHODS: Prostate stromal cells were stimulated with 10 nM DHT at different ages. The transcriptional activity of AR was detected by luciferase reporter assay. The secretion of AR regulatory factors FGF-2, KGF and IGF-1 were detected by Elisa. CCK8 detected with BPH-1 and LNCaP co-cultured BPH-1 and LNCaP proliferation. Immunohistochemistry, Q-PCR and Western blot were used to detect the expression of ARA55 in prostate stromal cells of different ages. The lentivirus interfered ARA55 and stimulated with 10nM DHT. The AR expression of stromal cells was detected by Western-blot. The AR transcriptional activity of stromal cells was detected by luciferase reporter assay Differences, Elisa test FGF-2, KGF, IGF-1 secretion differences. CCK8 detected the proliferation of BPH-1 and LNCaP after co-cultured with negative control group (ARA55-SC) and interference group (ARA55-sh) with BPH-1 and LNCaP. Results The stromal cells stimulated with 10 nM DHT showed higher AR transcriptional activity, AR-stimulated FGF-2, KGF, IGF-1 and up-regulation of epithelial cells (BPH-1 and LNCaP) . ARA55 expression in aged stromal cells is higher than that in young people. The expression of ARA55-sh in ARSCs was significantly higher than that in ARA55-SC AR cells (P <0.05) -1, LNCaP) (P <0.05), while there was no difference between ARA55.sh and ARA55-SC in young human stromal cells (P> 0.05). CONCLUSIONS: AR activity of stromal cells in the elderly is stronger than that of young people stimulated by 10nM DHT. Geriatric stromal cells may enhance AR transcriptional activity by ARA55 overexpression and AR regulation of cytokine secretion to promote the proliferation of BPH-1 and LNCaP.