Relationship between MMP7 expression and PTEN expression in TN tumors and cell lines, and MMP7 intensity correlation with PAM50 designation.

<p>Relationship between MMP7 expression and PTEN expression in TN tumors and cell lines (A and B), and MMP7 intensity correlation with PAM50 designation (C and D) are presented. <b>A</b>. Cell lines were downloaded from GEO (GD3285) with replicates averaged for each cell line. HCC38 and SUM149PT did not have replicates. Error bars represent the coefficient of variation between each of the replicates. MDA-MB231 and MCF7 cell lines had two distinct groups of replicates that are included on this graph to demonstrate the variability standard cell lines can have (most likely due to passage number differences). Relative levels of expression of MMP7 and PTEN mRNAs (published DASL intensities) from different TN cell lines showing a differential expression of high MMP7 in the TN (HCC38, HCC70, MDA-MB231, BT20, DKAT and SUM149T) cell lines as compared to non-TN cell lines (MCF7, T47D) as shown in the upper panel. Normalized mRNA levels for breast cancer cell lines show that the presence of a higher level of MMP7 mRNA in the individual TN cell lines is associated with relatively lower levels of PTEN mRNA. These reciprocal levels of expression of MMP7 to the PTEN mRNAs (DASL intensities) were observed in heat map of gene expression as shown in the lower panel. Cell line intensity data generated from Affymetrix Gene Expression arrays (Upper panel). RMA normalized intensity values shows relative inter-gene levels of each mRNA in a specific cell line with the (Lower panel) heat map using Z’ values calculated for each specific gene to visualize intra-gene mRNA levels. <b>B (i)</b>. Expression of MMP7 protein and the secretion of functional MMP7 were determined in the PTEN-null and PTEN expressing TNBC cells. High levels of expression of MMP7 protein and the secretion of functional MMP7 (casein zymogram from the conditioned media) in TNBC cells is inversely related to the presence of functional PTEN (protein expression as well as pAKT). <b>B (ii)</b>. Expression of MMP7 protein and the secretion of functional MMP7 were determined in the PTEN expressing non-TNBC cells (HR+ and HER2 over-expressed). <b>C</b>. Patient downloaded from GEO (GSE25055). Top panel showing PAM50 designations used to separate the patients into each subtype. Each subtype was then sorted from left to right by increasing MMP7 intensity. Bottom panel is sorted by increasing MMP7 from left to right. Bold vertical line separates pCR on the left with residual disease (RD) on the right. <b>D</b>. Heat Map of expression of mRNAs for MMP7, PTEN, CTNNB1, c-MYC genes (PAM50 data) in Basal, HER2E and HR+ subsets of BC. Table (Lower panel) shows the outcome data for triple negative breast cancer patients from PAM50 data set with high MMP7, and low MMP7. Patient data from <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0077425#pone-0077425-g006" target="_blank">Fig. 6C</a> were further examined for basal-like subtype using Z’ scores. Top panel shows increasing MMP7 Z’ score from left to right for all patient samples. Middle panel displays only patients with Z’ scores over 2.0 for MMP7, which shows the association with high MMP7 levels and low PTEN levels. The outcome data for the patients expressing low-MMP7 and high-MMP7 in their tumors are shown in the table (Lower panel).