36 research outputs found

    Array analysis of genes regulated by the PI3K/AKT pathway in lung epithelial cells.

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    <p>(A) Heatmap representation of expression values of DEGs comparing BEAS-AKT1-E17K, BEAS-PIK3CA-E545K and BEAS-shPTEN to BEAS-C control cells. Hierarchical clustering analysis was perfomed with Euclidean metric distance and Ward’s linkage rule. (B) The Venn diagram shows the number of common, specific and exclusive DEGs.</p

    Comparison analysis of Diseases and Bio Functions enriched in BEAS-2B cells expressing constitutively active PIK3CA, AKT1 or interfered for PTEN.

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    <p>IPA heatmap displays the top three Disease and Biofunctions enriched by DEGs that are common among BEAS-AKT1-E17K, BEAS-PIK3CA-E545K and BEAS-shPTEN cells (commons, rightmost column) or that are exclusive for each cell line (AKT1, PIK3CA, shPTEN). Diseases and Bio Functions were sorted for p-value, from the most significant (violet) to the least significant (white) values.</p

    The top 11 canonical signalling pathways influenced by constitutive PI3K signaling.

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    <p>Active PIK3CA (E545K)-expressing lentivirus was transduced in non-transformed lung epithelial cells (BEAS-2B). Transduced cells were selected in blasticydin, checked for expression of the exogenous PIK3CA-E545K, were analysed for their transcriptomes as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0030427#s2" target="_blank">Materials and methods</a>. <b>A.</b> Immunoblot for PIK3CA expression in transfected BEAS-2B cells. <b>B.</b> Heat map showing fold change patterns of DEGs induced by constitutive PI3K signalling. The heat map was generated in Matlab (Mathworks), and compares fold change patterns of DEGs in BEAS-2B-PI3K-E545K cells compared to parental BEAS-2B (p<0.01). Red: up-regulated genes; green: down-regulated genes. Fold changes of all down-regulated DEGs and all but one up-regulated DEG are #8 (central color spectrum bar). <b>C.</b> The top 11 functional categories determined by IPA, that were significantly up-regulated or down-regulated in BEAS-2B-PI3K-E545K cells compared to parental BEAS-2B are shown. The 2126 DEGs in BEAS-2B-PI3K-E545K were mapped to the IPA-defined network. The significance p-values that determine the probability that the association between the genes in the dataset and the canonical pathway is by chance alone were calculated by Fisher's exact test, and are expressed as −log (p-value). <b>D.</b> Bio-functions identified by IPA in the 2126 DEGs from BEAS-2B-PI3K-E545K compared with BEAS-2B. <b>E.</b> Sub-Categories and Functions identified through IPA showing the genes associated to lung cancer in the 2126 DEGs from BEAS-2B-PI3K-E545K compared with BEAS-2B.</p

    IHC and FISH analysis of PI3KCA in NSCLCs.

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    <p>A, left: SCC negative for PIK3CA expression; right: SCC positive for PIK3CA expression. B, left: ADC negative for PIK3CA expression; right: ADC positive for PI3KCA expression. Magnification 10× and 40×, respectively. C. Dual-color fluorescence in situ hybridization analysis of PIK3CA gene copy number. FISH analysis of PIK3CA (red signals) and chromosome region 3p14.1 (green signals). Left, NSCLC sample with diploid cells; right, NSCLC sample with multiple clustered spots of red signals of PIK3CA with 2 chromosome region 3p14.1 signals (gene amplification). Original magnification 100×.</p

    IHC and FISH analysis of AKT2 in NSCLCs.

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    <p>A, left: SCC negative for AKT2 expression; right: SCC positive for AKT2 expression. B, left: ADC negative for AKT2 expression; right: ADC positive for AKT2 expression. Magnification 10× and 40×, respectively. C. Dual-colour fluorescence in situ hybridization analysis of AKT2 gene copy number. FISH analysis of AKT2 (red signals) and chromosome region 19p13.1 (green signals). Left, NSCLC sample with diploid cells; right, NSCLC sample with multiple clustered spots of red signals of AKT2 with 2 chromosome region 19p13.1 signals (gene amplification). Original magnification 100×.</p

    Correlation between AKT activation and expression of the different members of the PI3K pathway in NSCLCs.

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    a<p>AKT activation was evaluated with as pS473 positivity and scored as negative (<10% of positive tumour cells) and high (>10% of positive tumour cells).</p>b<p>PIK3CA was graded as positive (>25% of tumour cells showed strong or diffuse immunopositivity) as moderate (>10% of tumour cells showed moderate immunopositivity) or negative (0–10% of the tumour cells showed weak, focal immunopositivity or absence of staining).</p>c<p>PTEN expression was classified as (+) when staining was detected in >50% of the cells, (+/−) when staining was detected in 25–50% of cells and (−) when staining was detected in 0–25% of cells. For statistical analysis PTEN expression was considered lost when samples were classified as (−).</p>d<p>AKT1 was graded as positive (>25% of positive tumour cells) as moderate (>10% of of positive tumour cells) or negative (0–10% of positive tumour cells).</p>e<p>AKT2 was graded as positive (>25% of positive tumour cells) as moderate (>10% of positive tumour cells) or negative (0–10% of positive tumour cells).</p>§<p>Statistically significant.</p
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