Chang G T, Blok L J, Steenbeek M, Veldscholte J, van Weerden W M, van Steenbrugge G J, Brinkmann A O
Department of Endocrinology and Reproduction, Erasmus University Rotterdam, the Netherlands.
Cancer Res. 1997 Sep 15;57(18):4075-81.
Differential gene expression between androgen-dependent (LNCaP-FGC) and androgen-independent (LNCaP-LNO) prostate cancer cells has been investigated using RNA arbitrarily primed and differential display PCR of mRNA. Four differentially expressed cDNA transcripts were identified, of which differential expression was confirmed by Northern blot analysis. Sequence analysis revealed two unknown (JC19 and GC79) and two known genes [B-cell translocation gene 1 and UDP-glucuronosyltransferase 2B15 (UGT2B15)]. JC19, GC79, and B-cell translocation gene 1 were more highly expressed in LNCaP-FGC cells compared with LNCaP-LNO cells, whereas UGT2B15 was only expressed in LNCaP-LNO cells. Androgens and 1,25-dihydroxyvitamin D3 were able to down-regulate UGT2B15 mRNA in LNCaP-LNO cells. For GC79 mRNA, down-regulation was only observed with androgens in LNCaP-FGC cells. Expression of JC19 mRNA was studied using a panel of human prostate cancer xenografts. In androgen-dependent xenografts, expression of JC19 mRNA was much higher compared with androgen-independent xenografts, in which significant expression was hardly detectable. The mRNA expression pattern in the xenografts is in good agreement with that observed in the cell culture system. In conclusion, the differential display technique used in the present study allows analysis of gene expression in vitro and in vivo and can be used for the identification of important genes involved in androgen-independent prostate cancer development.
利用RNA任意引物和mRNA差异显示PCR技术,研究了雄激素依赖性(LNCaP-FGC)和雄激素非依赖性(LNCaP-LNO)前列腺癌细胞之间的差异基因表达。鉴定出4种差异表达的cDNA转录本,其中差异表达通过Northern印迹分析得以证实。序列分析揭示了两个未知基因(JC19和GC79)和两个已知基因[B细胞易位基因1和UDP-葡萄糖醛酸基转移酶2B15(UGT2B15)]。与LNCaP-LNO细胞相比,JC19、GC79和B细胞易位基因1在LNCaP-FGC细胞中表达更高,而UGT2B15仅在LNCaP-LNO细胞中表达。雄激素和1,25-二羟基维生素D3能够下调LNCaP-LNO细胞中UGT2B15 mRNA的表达。对于GC79 mRNA,仅在LNCaP-FGC细胞中观察到雄激素诱导的下调。使用一组人前列腺癌异种移植模型研究了JC19 mRNA的表达。在雄激素依赖性异种移植中,JC19 mRNA的表达比雄激素非依赖性异种移植高得多,在后者中几乎检测不到明显表达。异种移植中的mRNA表达模式与细胞培养系统中观察到的模式高度一致。总之,本研究中使用的差异显示技术能够分析体外和体内的基因表达,可用于鉴定参与雄激素非依赖性前列腺癌发生的重要基因。