Endometrial carcinoma with ectopic human chorionic gonadotropin expression

• Aggressive course and treatment resistance characterize ectopic human chorionic gonadotropin. • Recurrence of endometrial cancer with ectopic hCG was treated with brachytherapy and EMACO. • The serum hCG level can serve as a marker in tumors with ectopic hCG expression

Wnt signaling in triple negative breast cancer is associated with metastasis

Background Triple Negative subset of (TN) Breast Cancers (BC), a close associate of the basal-like subtype (with limited discordance) is an aggressive form of the disease which convey unpredictable, and poor prognosis due to limited treatment options and lack of proven effective targeted therapies. Methods We conducted an expression study of 240 formalin-fixed, paraffin-embedded (FFPE) primary biopsies from two cohorts, including 130 TN tumors, to identify molecular mechanisms of TN disease. Results The annotation of differentially expressed genes in TN tumors contained an overrepresentation of canonical Wnt signaling components in our cohort and others. These observations were supported by upregulation of experimentally induced oncogenic Wnt/β-catenin genes in TN tumors, recapitulated using targets induced by Wnt3A. A functional blockade of Wnt/β-catenin pathway by either a pharmacological Wnt-antagonist, WntC59, sulidac sulfide, or β-catenin (functional read out of Wnt/β-catenin pathway) SiRNA mediated genetic manipulation demonstrated that a functional perturbation of the pathway is causal to the metastasis- associated phenotypes including fibronectin-directed migration, F-actin organization, and invasion in TNBC cells. A classifier, trained on microarray data from β-catenin transfected mammary cells, identified a disproportionate number of TNBC breast tumors as compared to other breast cancer subtypes in a meta-analysis of 11 studies and 1,878 breast cancer patients, including the two cohorts published here. Patients identified by the Wnt/β-catenin classifier had a greater risk of lung and brain, but not bone metastases. Conclusion These data implicate transcriptional Wnt signaling as a hallmark of TNBC disease associated with specific metastatic pathways

Expression of MMP7, one of the transcriptional targets of the WP as compared to other members of the MMP family in different subtypes of breast tumor samples from the Montreal and Georgia cohorts.

<p>(<b>A</b>) Figure shows MMP7 expression in the Montreal cohort. Expression of MMP7 is higher in the TN subtype. Expression is sorted from the highest to the lowest level for TN (black), HER2+ (grey), and luminal (white) breast tumor subtypes as classified by IHC. Inset shows average expression of MMP7 by subtype in the cohort. Average expression of MMP7 is significantly higher (P< 0.05) in the TN (black) subtype as compared to both the HER+ (grey) and luminal (white) subtypes as indicated by the asterisk. Numbers within bracket are the patients in each subtype. (<b>B</b>) Figure shows selective upregulation of mRNA for MMP7 in TN tumors of the Montreal cohort as evident from the DASL assay average intensity data for different metalloproteinases. Expression of MMP1, MMP2, MMP3, MMP9, MMP11 and MMP7 grouped by hormone receptor and HER2 status; HR+/HER2-, HER2+, and TN. HR+ (hormone receptor+) is ER+ and/or PR+; HER2+ is HER2-positive and HR±; TN is triple negative. Error bars represent STDEV. Level of transcript expression is represented by DASL average intensity. *P< 0.05, and n represents number of samples in each subset. (<b>C</b>) Figure shows the expression of MMP7 with respect to IHC subtypes in two of our data sets (Montreal-91 [GSE17650], Georgia-137 [GSE18539]) and another published data set (MSK-96 [GSE2603]). The expression of mRNA for MMP7 is significantly upregulated in TN as compared to the luminal subtype (HR+) in all the sets and HER2+ subtype in the Georgia-137 cohort (p = 6.00×10^-22). The expression of the mRNA for the gene is significant in univariate analysis (p < 0.01). (<b>D</b>) The upregulation of MMP7 mRNA is observed in a subset of TNBC specimens as shown by the bar graph of the expression of MMP7 in each tumor in the cohort. Bars represent MMP7 mRNA expression in 87 FFPE breast tumor specimens (36 HR+/HER2- tumors, 27 HER2+/ HR± tumors and 24-TN tumors from the Montréal cohort) grouped by hormone receptor and HER2 status. MMP7 expression was upregulated in a subset ~40% (9/24) of TN tumor samples (shown in red) but not in any of the other (0/63) tumor samples (shown in blue).</p

Expression of mRNA, protein and enzymatic activity of MMP7 in TNBC cell lines as compared to HER2+ and luminal BC cell lines.

<p>(<b>A</b>) Bar diagram (upper panel) shows mRNA expressions for MMP7 in HCC70 (black), MDA-MB468 (grey), and MCF7 (white) BT-like cell lines. Error bars represent one standard error of the mean and p-values are determined using a t-test for unequal sample sizes and means. Bar diagram in the lower panel (left) shows the validation of the expression of mRNA for MMP7 by RT-PCR in the BT cell lines. Expression of MMP7 measured by RT-PCR is concordant with microarray data for the HCC70, MDA-MB-468, and MCF7 cell lines. Expression is determined by the average of three replicates and bars represent one standard error of the mean (*p < 0.05). Bar diagram in the lower panel (right) shows RT-PCR expression of mRNA for MMP7 in different BT cell lines (MCF7, SKBr3, HCC38, SUM149). Values represent the mean of three replicates, and error bars represent one standard error of the mean (*p < 0.05). (<b>B</b>) MMP7 mRNA expression was upregulated in only a subset of TN breast cancer cell lines (shown in red) and not in any other types of BC-like cell lines. MMP7 affymatrix expression data from Neve et al. [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0077425#B36" target="_blank">36</a>], grouped by hormone receptor and HER2 status. (<b>C</b>) Immunoblots showing differential expression of MMP7 protein (lanes 5-7, 12, 13, 14–16, 18-19) in TN breast cancer (T) cell lines as compared to HER2+ (H) and luminal (L) BC-like cell lines. Levels of MMPs 2, 9 and 14 are determined as references. Inset shows a differential expression of expression of MMP7 protein in a subset of TN breast cancer cell lines as determined by densitometry semi-quantitative analyses (imageJ program; absorbance ratio of MMP7 to beta-actin) of respective immunoblots. (<b>D</b>) Caesin-zymogram of the secreted-MMP7 and corresponding immunoblots of the cellular-MMP7 protein levels show both an increased expression and enzymatic activity of MMP7 (lanes 3, 5, 6, 9, 10) in TN breast cancer cell line (T) as compared to HER2+ (H) and luminal (L)-like cell lines.</p