Application of RNA-Seq transcriptome analysis: CD151 is an Invasion/Migration target in all stages of epithelial ovarian cancer

Rebecca A Mosig,Maurizio D'Incalci,Hardik Shah,Sergio Marchini,Robert Fruscio,Emir Senturk,Li Lin,Samantha Cohen,Peter Dottino,Ravi Sachidanandam,John A Martignetti,Fei Huang,Peter Schlosshauer

doi:10.1186/1757-2215-5-4

Abstract

BackgroundRNA-Seq allows a theoretically unbiased analysis of both genome-wide transcription levels and mutation status of a tumor. Using this technique we sought to identify novel candidate therapeutic targets expressed in epithelial ovarian cancer (EOC).MethodsSpecifically, we sought candidate invasion/migration targets based on expression levels across all tumors, novelty of expression in EOC, and known function. RNA-Seq analysis revealed the high expression of CD151, a transmembrane protein, across all stages of EOC. Expression was confirmed at both the mRNA and protein levels using RT-PCR and immunohistochemical staining, respectively.ResultsIn both EOC tumors and normal ovarian surface epithelial cells we demonstrated CD151 to be localized to the membrane and cell-cell junctions in patient-derived and established EOC cell lines. We next evaluated its role in EOC dissemination using two ovarian cancer-derived cell lines with differential levels of CD151 expression. Targeted antibody-mediated and siRNA inhibition or loss of CD151 in SKOV3 and OVCAR5 cell lines effectively inhibited their migration and invasion.ConclusionTaken together, these findings provide the first proof-of-principle demonstration for a next generation sequencing approach to identifying candidate therapeutic targets and reveal CD151 to play a role in EOC dissemination.

Highlights

RNA-Seq allows a theoretically unbiased analysis of both genome-wide transcription levels and mutation status of a tumor
CD151 is expressed across all stages of epithelial ovarian cancer (EOC) A total of 16 papillary serous epithelial ovarian tumors, representing early- and late-stage disease and metastatic nodules and recurrences, and two borderline ovarian tumors were selected for RNA-Seq in our “discovery set” of samples
CD151 is expressed in EOC tumors and normal ovary surface epithelial cells Having identified overexpression on the RNA level, we evaluated the expression of CD151 protein in tumors and normal ovary tissues using immunohistochemistry

Summary

Introduction

RNA-Seq allows a theoretically unbiased analysis of both genome-wide transcription levels and mutation status of a tumor Using this technique we sought to identify novel candidate therapeutic targets expressed in epithelial ovarian cancer (EOC). Since the introduction of platinum-based drugs as first line chemotherapy in the early 1980’s followed by the addition of taxane containing agents in the mid-1990’s, there has been little change to the first line treatment of EOC [3]. Novel administration methods, such as intraperitoneal therapy, and dosing, such as dose-dense taxol, have yielded slight improvements in progression-free survival and overall survival [3]. The need for new targets and drugs remains high [1]

Methods

Results

Conclusion