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. 2009 Jan;174(1):276–286. doi: 10.2353/ajpath.2009.080086

Figure 1.

Figure 1

IRS2 is elevated in Pten+/− neoplasia of the prostate and uterus and in tumor cell lines. A: Immunohistochemical analysis of IRS2 protein expression in human prostate cancer (a), adjacent area of hyperplasia (b), human endometrial cancer (c), and adjacent proliferative endometrium (d). IRS2 expression is increased in cancer relative to nonneoplastic adjacent epithelium. Basal epithelial cells of the prostate and proliferative endometrium show light IRS expression (black arrows). B: Protein lysates were generated from human tumor cell lines under recommended growth conditions and immunoblots were examined for IRS2 protein and tubulin. IRS2 protein levels varied greatly among lines. IRS2 protein levels were relatively low in MCF10A (immortal normal breast cell line), MDA-MB-468, BT549, and LNCaP, and much higher in HCC1143, DBTRG, PC3, and DU145. Cell lines not shown also had low levels of IRS2 protein. FISH of the lines showed that HCC1143 and DBTRG had increased copies of the IRS2 gene (red), whereas DU145 has two copies (arrows). Probing of PC3 also showed normal IRS2 copy number (data not shown). A RB FISH probe (green) was used as a control because it is also on chromosome 13q. Metaphase spreads are shown that indicate that HCC1143 has a homogeneously staining region (yellow arrow) for IRS2 in addition to two other copies (white arrows) and that DBTRG has at least seven copies of IRS2, five of which are attributable to an increase in the number of copies of chromosome 13q. PTEN genotypes: MCF10A wild type (+/+); DU145 heterozygous mutant (+/−); HCC1143 wild type (+/+); MDA-MB-468, BT549, DBTRG, PC-3, LNCaP both alleles mutated and/or deleted (−/−). Original magnifications, ×400 (A).