Abstract DPOC-003: DIFFERENTIAL PROTEIN EXPRESSION ANALYSIS OF SV40–IMMORTALIZED OVARIAN SURFACE, SV40–IMMORTALIZED FALLOPIAN TUBE AND OVARIAN CANCER SUBTYPE CELL LINE SECRETOMES BY ITRAQ®
Authors: Zhou Yaling , Brooks Daniel , Dey Sumana , Higgins LeeAnn , Markowski Todd , Hamilton Chad , Howard Robin , Raiciulescu Sorana , Skubitz Amy , Andersen John D. 
Published: June 2017

BACKGROUND: In 2016, an estimated 22,280 new cases of ovarian cancer will be diagnosed with more than 14,000 women succumbing to the malignancy. These figures have not changed in the last decade. During this time new ovarian cancer models have been refined that involve either the fallopian tube or ovarian surface epithelial cells as the origin for tumorigenesis. With refined proteomics methodology, identification of new secreted biomarkers may be revealed that are specific to a primary fallopian tube or ovarian surface epithelium origin and may ultimately aid in the diagnosis, prognosis or therapy of ovarian cancer patients.

EXPERIMENTAL PROCEDURES: The secretomes of eight cell lines, represented by SV40 immortalized fallopian tube (FT-194, FT-190), SV40 immortalized ovarian surface epithelium (1816-575), low-grade serous adenocarcinoma (HEY), high-grade serous adenocarcinoma (TOV-1946), clear cell adenocarcinoma (TOV21G), cisplatin-sensitive high-grade serous adenocarcinoma (A2780-S) and cisplatin-resistant high-grade serous adenocarcinoma (A2780-CP), were submitted for isobaric tagging for relative and absolute quantification (iTRAQ®) differential expression analysis. Briefly, each of the eight secretomes were trypsin-digested and labeled with one of eight iTRAQ® 8-plex reagents and subsequently submitted for either two-dimensional capillary liquid chromatography direct data dependent peptide tandem mass spectrometry on an Orbitrap Velos system or one-dimensional mass spectrometry analysis.

RESULTS: 456 proteins were identified with the one-dimensional analysis with 2 distinct peptides. 2069 distinct differentially expressed proteins were identified in the two dimensional analysis. Initial relative differential expression rate comparisons of control cell lines, FT-194, FT-190 and 1816-575 versus ovarian cancer subtype and therapy-associated cell lines, HEY, TOV1946, TOV21G, A2780S and A2780CP, are promising. Four proteins were increased greater than three-fold across all ovarian cancer cell lines compared to normal fallopian tube and ovarian surface epithelial control cell lines: stromelysin-1, pigment epithelium-derived factor, matrix metalloproteinase-9 and receptor expression-enhancing protein 6. Large scale inferential statistical methodology will be used to differentiate cell lines.

Citation Format: Yaling Zhou PhD, Daniel Brooks, PhD, Sumana Dey PhD, LeeAnn Higgins, PhD, Todd Markowski, BSc, Chad Hamilton MD, Robin Howard, MA, Sorana Raiciulescu MSc, Amy Skubitz PhD, John D. Andersen DO. DIFFERENTIAL PROTEIN EXPRESSION ANALYSIS OF SV40–IMMORTALIZED OVARIAN SURFACE, SV40–IMMORTALIZED FALLOPIAN TUBE AND OVARIAN CANCER SUBTYPE CELL LINE SECRETOMES BY ITRAQ® [abstract]. In: Proceedings of the 11th Biennial Ovarian Cancer Research Symposium; Sep 12-13, 2016; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2017;23(11 Suppl):Abstract nr DPOC-003.