Dysregulation of AKT3 along with a small panel of mRNAs stratifies high-grade serous ovarian cancer from both normal epithelia and benign tumor tissues.

Screening methods of High-Grade Serous Ovarian Cancer (HGSOC) lack specificity and sensitivity, partly due to benign tumors producing false-positive findings. We utilized a differential expression analysis pipeline on malignant tumor (MT) and normal epithelial (NE) samples, and also filtered the results to discriminate between MT and benign tumor (BT). We report that a panel of 26 dysregulated genes stratifies MT from both BT and NE. We further validated our findings by utilizing unsupervised clustering methods on two independent datasets. We show that the 26-genes panel completely distinguishes HGSOC from NE, and produces a more accurate classification between HGSOC and BT. Pathway analysis reveals that AKT3 is of particular significance, because of its high fold change and appearance in the majority of the dysregulated pathways. mRNA patterns of AKT3 suggest essential connections with tumor growth and metastasis, as well as a strong biomarker potential when used with 3 other genes (PTTG1, MND1, CENPF). Our results show that dysregulation of the 26-mRNA signature panel provides an evidence of malignancy and contribute to the design of a high specificity biomarker panel for detection of HGSOC, potentially in an early more curable stage.

Downregulation of HOXC6 in Serous Ovarian Cancer.

Homeobox (HOX) genes encode transcription factors critical to morphogenesis and cell differentiation. Although dysregulation of several HOX genes in ovarian cancer has been reported, little is known about HOXC6 expression in epithelial ovarian cancer. In this report, analysis of laser capture microdissected samples determined HOXC6 expression patterns in normal versus malignant serous ovarian carcinoma tissues. HOXC6 protein was quantified by ELISA in parallel serum samples and further validated in a larger cohort of serum samples collected from women with and without serous ovarian carcinoma. These data demonstrate significant downregulation of HOXC6 in serous ovarian cancer.

Molecular evaluation of proliferative-phase endometrium may provide insight about the underlying causes of infertility in women with endometriosis.

PURPOSE: To determine if endometrial gene expression is different in women with endometriosis-related infertility and fertile women. METHODS: Prospective study of mid-follicular phase endometrium in 47 subjects in two phases: microarray study of 10 infertile women with endometriosis and five fertile controls, and a quantitative real-time PCR (qRT-PCR) study of 27 infertile women with endometriosis and 15 fertile controls. Gene expression was determined by DNA microarray, and qRT-PCR used for 12 "promising" genes based on the microarray analysis. RESULTS: Compared to fertile controls, women with stage I-II endometriosis had 23, and women with stage III-IV had 35 genes that were significantly up- or down-regulated by microarray. However, using qRT-PCR, only chemokine ligand (CXCL) 13 was significantly down-regulated and somatostatin was significantly up-regulated with early endometriosis, and only CXCL 14 was significantly down-regulated with advanced endometriosis compared to fertile controls. CONCLUSIONS: Our findings indicate that the pattern of gene expression in proliferative-phase endometrium is different when comparing tissue from patients with endometriosis versus fertile controls. Recognition of these endometrial alterations could be helpful to diagnose and stage endometriosis, and may provide insight to explain why conception rates are low in women with endometriosis.

Correlation analysis of HOX, ErbB and IGFBP family gene expression in ovarian cancer.

Utilizing microarray gene expression data in cancer research possesses the ability to identify deregulated cellular pathways involved in malignant development. This study investigated the relationships of three gene families, HOX, ErbB and IGFBP, with regard to the development of ovarian cancer. These families were of interest because of similar chromosomal locations and their deregulated expression in ovarian cancer. Higher level statistics were used to differentially analyze microarray data in 65 ovarian samples to assess correlation and relationships among the gene families of interest. Fifteen genes in the three families were found to be significantly deregulated. Thirty-eight significant correlations were found within and between the genes of interest. Our data indicates that the significantly modeled relationships between HOX, ErbB and IGFBP gene pairs could provide insight into the underlying biological mechanisms in ovarian cancer.

Gene expression profiling of ovarian tissues for determination of molecular pathways reflective of tumorigenesis.

Ovarian cancer is the fourth leading cause of gynecological cancer death among women in the United States. Early detection is a critical prerequisite to initiating effective cancer therapy. Gene microarray technology and proteomics have provided much of the biomarkers with potential use for diagnosis. However, more research is needed to fully understand disease onset and progression. To this end, we have performed microarray analysis with the goal of identifying molecular interaction networks defining tumor growth. Microarray analysis was performed on a limited set of ovarian tissues with various pathological diagnoses using Human Genome Focus Array (HGFA) for the detection of approximately 8500 human transcripts. Hierarchical clustering identified groups of ovarian tissues reflective of low malignant potential/early cancer onset and possible pre-cancerous stages involving small molecule, cytokine and/or hormone-dependent feed-back responses specific to the pelvic reproductive system and a priori initiated tumor suppression mechanisms. ANOVA followed by post hoc Scheffe confirmed our hypotheses. Moreover, we established a protein/protein interaction database associated with HGFA probe sets. This database was used to build and visualize molecular networks integrating small but significant changes in gene expression. In conclusion, we were able for the first time to delineate an intersecting genetic pattern linking ovarian tissues reflective of low potential malignancy/early cancer onset stages via long distance signaling between tissues of gynecological origin.

Estrogen receptor-alpha messenger RNA variants that lack exon 5 or exon 7 are coexpressed with wild-type form in human endometrium during all phases of the menstrual cycle.

OBJECTIVE: We have assessed the expression levels of messenger RNA for estrogen receptor-alpha and splice variants lacking exon 5 or exon 7 that presumably exert dominant positive (splice variants lacking exon 5) and negative (splice variants lacking exon 7) effects, respectively, on estrogen responses in the human endometrium. STUDY DESIGN: This was a prospective study that was conducted at an academic community-based hospital. The patients, aged 18 to 40 years, underwent hysterectomy for benign gynecologic causes. Eighty-one endometrial specimens (46 proliferative, 35 secretory) were analyzed with the use of reverse transcription-polymerase chain reaction assay for the messenger RNA levels of estrogen receptor-alpha, and splice variants lacking exon 5 and exon 7. RESULTS: Wild-type estrogen receptor-alpha and splice variants splice variants lacking exon 5 and lacking exon 7 messenger RNAs were detected in all endometrial specimens throughout the menstrual cycle. In addition, a double-splice estrogen receptor-alpha messenger RNA variant (splice variants lacking exon 5 and exon 7) was detected at constant low levels of expression. Semiquantitative analysis showed higher levels of estrogen receptor-alpha messenger RNA in the early and mid proliferative endometrial phases than in late proliferative and secretory endometrium ( P <.05). The splice variant lacking exon 7 messenger RNA expression level was about 10-fold higher than the splice variant lacking exon 5 messenger RNA relative to wild-type estrogen receptor-alpha messenger RNA ( P <.001). The expression of splice variants lacking exon 5 compared with wild-type estrogen receptor-alpha messenger RNA is relatively constant throughout endometrial development. In contrast, an examination of the ratio of the levels of splice variants lacking exon 7 to wild-type estrogen receptor-alpha messenger RNA indicated a small, but significantly higher, splice variant lacking exon 7 level in the mid secretory phase (postovulatory days 5-8) than the mid proliferative and early secretory phases ( P <.05). CONCLUSION: We found no evidence of differential coexpression of the positive dominant estrogen receptor variant, splice variants lacking exon 5, with wild-type estrogen receptor-alpha. We did find that the dominant negative splice variant lacking exon 7 was slightly increased relative to wild-type estrogen receptor-alpha in the postovulatory phase. Future investigation is required to suggest the biologic significance of the observed increased relative expression of the splice variants lacking exon 7 in secretory endometrium and to determine the function of splice variants lacking exon 5 and splice variants lacking exon 7.