Global gene expression of histologically normal primary skin cells from BCNS subjects reveals “single-hit” effects that are influenced by rapamycin

The study of dominantly heritable cancers has provided significant insights about tumor development. Basal cell nevus syndrome (BCNS), or Gorlin syndrome (OMIM #109400), is an autosomal dominant inherited disorder that is characterized by developmental abnormalities and postnatal occurrence of various neoplasms, principally basal cell carcinomas (BCCs). We performed an exploratory gene expression profiling of primary cultures, including keratinocytes and fibroblasts derived from clinically unaffected skin biopsies of BCNS gene-carriers (PTCH1 +/-) and normal individuals. Low pass Fibroblast and Keratinocyte cell lines from study participants were treated with 0 ng/ml, 10 ng/ml, and 50 ng/ml of Sirolimus for 48 hours. The RNA was extracted using our standard Guanidine Thiocyanate Procedure and cleaned using the Qiagen RNeasy Micro Kit (according to the manufacturer’s directions). We then analyzed the RNA on Affymetrix U133 Plus 2.0 expression arrays. Description of Sample titles: Example 1: 01F48S10 = participant 01, fibroblasts, 48-hour treatment with sirolimus, dose 10 ng/ml. Example 2: 01K48S50 = participant 01, keratinocytes, 48-hour treatment with sirolimus, dose 50 ng/ml. Example 3: 01Fib = participant 01, fibroblasts, no treatment with sirolimus.