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Figure 1 | Molecular Cancer

Figure 1

From: microRNA regulation of mammalian target of rapamycin expression and activity controls estrogen receptor function and RAD001 sensitivity

Figure 1

mTORC1 activation is Associated with an ER Negative Breast Tumor Phenotype. Deep sequencing data obtained from TCGA data portal was analyzed with respect to ERα (either ERα positive or negative) status for expression of (A) mTOR associated gene sets (mTOR, TSC1, TSC2, Rheb, Rictor, Raptor, Rheb) and (B) miR-155 host gene (HG) expression. Heat mat depicts high expression (red, +1) and low expression (green, −1). Platform analyzed was TCGA breast invasive carcinoma gene expression IlluminaHiSeq n = 1032. Analysis was sorted based on ERα gene Signature where positive for ER = orange and negative for ER = blue. (C) MCF-7-vector and –miR-155 cells were harvested for qPCR analysis for mTOR associated genes Rictor, TSC1, Deptor, Rheb, Raptor, and p70s6 kinase. Ct values were normalized to β-actin and MCF-7-vector cells designated as 1. * p < 0.05 for n ≥ 3. (D) MCF-7-vector and –miR-155 cells were harvested for western blot analysis for total and phospho-mTOR (S2448) and mTOR associated proteins Rictor, Raptor, TSC1, and p-70 s6 kinase. Expression normalized to RHO-GDIα, n ≥ 3. (E) MCF-7-vector and miR-155 cells were harvested for western blot analysis for phosphorylation of downstream mTORC1 associated proteins p-eIF4B (S422), p-S6 ribosomal protein (S235/236), p-p70s6 kinase (Thr389), p-eEF2K (S366). Expression was normalized to RHO-GDIα, n = 2. (F) qPCR of MCF-7-vector and miR-155 cells for downstream mTORC2 regulated gene PKCα, normalization was to β-actin and vector designated as 1, n = 3.

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