Sinisi A A, Bellastella A, Prezioso D, Nicchio M R, Lotti T, Salvatore M, Pasquali D
Istituto di Endocrinologia, Facoltà di Medicina, Seconda Università di Napoli, Italy.
J Clin Endocrinol Metab. 1997 Aug;82(8):2566-9. doi: 10.1210/jcem.82.8.4142.
The transcripts of five SRIH receptor subtypes (SSTR1, SSTR2, SSTR3, SSTR4, and SSTR5) were investigated by RT-PCR in epithelial cells (EC) and stromal cells (SC) from primary cultures of five normal human prostates and six prostate cancers. Primary cultures of prostate EC were established in serum-free keratynocyte medium with 5% FCS, epidermal growth factor, and bovine pituitary extract; SC were cultured in MEM with 10% FCS. Total RNA was extracted from EC and SC using a modified guanidine thiocyanate method. RT-PCR was performed after deoxyribonuclease treatment, using SSTR1-, SSTR2-, SSTR3-, SSTR4-, and SSTR5-specific-primers and adding glyceraldehyde-3-phosphate dehydrogenase-specific primers as internal control. A PCR product of the expected size of 334 bp, corresponding to SSTR1, was expressed only in EC from prostate cancer, whereas the expected 461-bp product of SSTR2 was found only in EC from normal prostate. SSTR3 messenger RNA was undetectable in normal and cancer EC, whereas SSTR4 and SSTR5 were present in both cell types. SSTR1, SSTR2, SSTR3, SSTR4, and SSTR5 messenger RNAs were not expressed in SC from both normal and cancer prostates. The RT-PCR method clearly demonstrated SSTRs' expression in the human prostate EC in vitro with differences between normal and tumoral samples. Our results may explain the ineffectiveness of some SSTR2 selective SRIH analogues in the treatment of prostate cancer and suggest that the absence of SSTR2 could represent a growth advantage in prostate cancer.
采用逆转录聚合酶链反应(RT-PCR)对来自5例正常前列腺和6例前列腺癌原代培养的上皮细胞(EC)和基质细胞(SC)中5种生长抑素受体亚型(SSTR1、SSTR2、SSTR3、SSTR4和SSTR5)的转录本进行了研究。前列腺EC的原代培养在含有5%胎牛血清(FCS)、表皮生长因子和牛垂体提取物的无血清角质形成细胞培养基中进行;SC在含有10% FCS的MEM培养基中培养。使用改良的异硫氰酸胍法从EC和SC中提取总RNA。在脱氧核糖核酸酶处理后进行RT-PCR,使用SSTR1、SSTR2、SSTR3、SSTR4和SSTR5特异性引物,并添加甘油醛-3-磷酸脱氢酶特异性引物作为内对照。预期大小为334 bp的对应于SSTR1的PCR产物仅在前列腺癌的EC中表达,而预期大小为461 bp的SSTR2产物仅在正常前列腺的EC中发现。在正常和癌性EC中均未检测到SSTR3信使核糖核酸,而SSTR4和SSTR5在两种细胞类型中均存在。正常和癌性前列腺的SC中均未表达SSTR1、SSTR2、SSTR3、SSTR4和SSTR5信使核糖核酸。RT-PCR方法清楚地证明了生长抑素受体在人前列腺EC中的体外表达,且正常和肿瘤样本之间存在差异。我们的结果可能解释了一些SSTR2选择性生长抑素类似物在前列腺癌治疗中无效的原因,并表明SSTR2的缺失可能代表前列腺癌的生长优势。