Development of gene expression signatures for MCF10A cell lines carrying HER2 and WT or H1047R PIK3CA oncogenes.

We wanted to know how introduction of H1047R oncogenic mutation of PIK3CA (p110a subunit of PI3K) in HER2-overexpressing MCF10A human non-transformed mammary epitheial cell lines introduces change in gene expression levels. Our control was HER2-overexpressing MCF10A cell line with wild-type (WT) PIK3CA. Mutated oncogene-induced gene expression signature was measured and compared with that induced by the WT oncogene from cell lines harvested at 70-80% confluency. MCF10A/HER2/WT [C] and MCF10A/HER2/HR [T] PIK3CA cell lines from two different passages (p18 and p20) were grown in media containing DMEM/F12, 5% horse serum, 20ng/ml EGF, 100ng/ml cjolera toxin, 10mg/ml insulin, 0.5mg/ml hydrocortisone. Total RNA was isolated by TRIzol method when cells were 70-80% confluent.