Characterization of a method for profiling gene expression in cells recovered from intact human prostate tissue using RNA linear amplification

摘要: Coupling array technology to laser capture microdissection (LCM) has the potential yield gene expression profiles of specific cell populations within tissue. However, remaining problems with linear amplification preclude accurate profiling when using low nanogram amounts RNA recovered after LCM human We describe a novel robust method reliably amplify LCM, allowing direct probing 12K arrays. The fidelity was demonstrated by comparing ability amplified (aRNA) versus that native identify differentially expressed genes between two different lines, demonstrating 99.3% concordance observations. Array findings were validated quantitative polymerase chain reaction analysis randomly selected subset 32 genes. Using recover normal (N=5 subjects) or cancer (N=3) from intact prostate tissue, three identified. Independent investigators have previously identified differential these genes, hepsin and beta-microseminoprotein, in cancer. Taken together, current study demonstrates can readily be performed on present complex It also this approach efficiently identifies biologically relevant