Microarray screening of RNA samples from low-, intermediate- and high-passage human prostate LNCaP cells

This laboratory studies the structures, biosynthesis, and functions of mucin glycans The low-(<33) and high-(>80)passage human prostate LNCaP cells developed by Dr. Ming-Fong Lin (Lin et al, Urology 166:1943, 2001) have been considered as the best in vitro model that reflects the progression of prostate cancer from androgen-dependent to androgen-independent stage (Parajuli et al, Cancer Res. 61:8227, 2001; Demeade et al, The Prostate 54:249, 2003; Unni et al, Cancer Res. 64:7156, 2004). The high-passage cells are metastatic while the low-passage cells are not. RT-PCR analysis of these two cell clones for 25 glycogenes among 8 different groups of glycogenes showed that only high-passage cells expressed two (isozymes 1 and 5) of the five known GlcNAc6ST genes. In addition, P-selectin blot analysis showed qualitative and quantitative differences between these clones. They included 3 higher intensity bands and 3 lower intensity bands in high-passage cells as compared to low-passage cells. The difference in P-selectin-specific glycoconjugates between low and high-passage LNCaP cells should help identify the glycans that distinguish high- from low-passage LNCaP cells. RNA preparations from low-, intermediate-, and high-passage human prostate LNCaP cells were sent to Microarray Core (E). *Concurrent analysis of glycan profiles in these cells was performed in Core C*. Three replicate samples from each condition were used in the study. The RNA was amplified, labeled, and hybridized to the GLYCOv3 microarrays. Data was analyzed to identify the glycogenes, glycans, and/or carbohydrate binding proteins responsible for the difference in metastatic property between low- and high-passage LNCaP cells. The intermediate-passage cells were used for identifying changes in glycogene, glycan, and/or carbohydrate binding protein expression that may be responsible for the earliest change of the metastatic property.