Open Access Research Article
Elamin et al., J Cancer Sci Ther 2017, 9:10
DOI: 10.4172/1948-5956.1000494
J Cancer Sci Ther, an open access journal
ISSN: 1948-5956
Volume 9(10) 705-708 (2017) - 705
Journal of
Cancer Science & Therapy
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ISSN: 1948-5956
Keywords: Breast cancer; Tumor markers; Proteomics; 2D-PAGE;
Mass spectrometry
Abbreviations: 2DE: Two-Dimensional Gel Electrophoresis; α1-
AT: Alpha-1 Anti-Trypsin; DTT: Dithiothreitol; Pin1: Peptidylprolyl
Cis-Trans Isomerase; PMF: Peptide Mass Fingerprinting; PrdxV:
Peroxiredoxin V; ROS Reactive Oxygen Species; SDS PAGE: Sodium
Dodecyl SulfatePolyacrylamide Gel Electrophoresis; MALDI TOF:
Matrix Assisted Laser Desorption Ionization - Time of Flight; Mass
Spectrometry; IPG: Immobilized pH Gradient; IEF: Iso-Electric
Focusing
Introduction
Worldwide, breast cancer (BC) ranks as the top cancer among
women, accounting for 23% of the total cancer cases and the principal
cause of cancer death (14%) among females [1]. In Sudan, breast
cancer is considered as the most frequent hospital treated malignancy,
accounting for about one-ffh of all cancers reported in both sexes
[2]. Among Sudanese women, breast and cervical Cancers account for
about 50% of all cancers [3].
Proteomic techniques are powerful tools to help in the identifcation
of tumor markers. Tese markers are thought to be helpful in the
clinical management of cancer patients, assisting in diagnostic,
staging, evaluation of therapeutic response, detection of recurrence
and metastasis, and the development of new treatment modalities [4].
Tere are few breast tumor markers being used routinely in clinics,
such as follow up molecular markers (CA 15-3, CA 27.29 and CEA),
markers of prognostic and/or therapeutic value (hormone receptors,
HER-2, Ki-67, p53 proteins), and the genes for hereditary breast cancer
[5]. Nevertheless, the number of biomarkers reaching routine clinical
use remains unacceptably low. Terefore, the search is still ongoing
for biomarkers to improve early and specifc detection and disease
monitoring [6]. Te present study aims to investigate the protein
profles of a panel of Sudanese breast cancer tissues and controls, with
the target of identifying proteins that could potentially be used as
tumor markers.
*Corresponding author: Muntaser E. Ibrahim, Institute of Endemic Diseases,
University of Khartoum, Sudan, Tel: 24911793267; E-mail: mibrahim@iend.org
Received October 17, 2016; Accepted October 25, 2017; Published October 27,
2017
Citation: Elamin A, Franz T, Ibrahim ME (2017) Identifcation of Potential Tumor
Markers in Sudanese Breast Cancer Patients Using a Proteomic Approach. J
Cancer Sci Ther 9: 705-708. doi:10.4172/1948-5956.1000494
Copyright: © 2017 Elamin A, et al. This is an open-access article distributed under
the terms of the Creative Commons Attribution License, which permits unrestricted
use, distribution, and reproduction in any medium, provided the original author and
source are credited.
Identification of Potential Tumor Markers in Sudanese Breast Cancer
Patients Using a Proteomic Approach
Amany Elamin
1,2
, Thomas Franz
3
and Muntaser E Ibrahim
1
*
1
Institiute of Endemic Diseases, University of Khartoum, Sudan
2
Commission for Biotechnology and Genetic Engineering, National Center for Research, Sudan
3
Proteomics Core Facility, European Molecular Biology Laboratory (EMBL), Heidelberg, Germany
Abstract
Breast cancer (BC) is considered as a major health problem in Sudan, being the most frequent hospital
treated malignancy and the commonest cancer affecting Sudanese females (34%). Proteomics provides tools
that investigate the precise molecular defect(s) in breast cancer tissue. Moreover, it plays a growingly important
role in tumor markers discovery. In this study, a proteomic assay (2D-PAGE) was used to investigate the protein
profles of a panel of 12 Sudanese breast malignant tissues and 12 matched controls. Protein spots of interest were
excised manually, and in-gel digested using trypsin. Proteins were identifed using mass spectrometry (MALDI-
TOF). Mascot program was used to search protein databases for matching peptides from known proteins. The
overall profle was relatively similar, the preliminary results identifed three proteins to be differentially expressed,
suggesting that they may perform a role in breast neoplasia. Although the number of samples investigated in this
study is comparatively small, to allow authoritative conclusions, the study provided three proteins that might be
potential tumor markers in Sudanese breast cancer patients requiring further investigations and validation.
Materials and Methods
Tis is a case-control hospital-based study. A panel of 12 Sudanese
breast cancer patients (12 tumor-control pairs) were recruited in this
study. Tissue biopsies were collected by Breast Cancer Research Group
members at the Institute of Endemic Disease, University of Khartoum,
from hospitals around Khartoum State. Both fresh normal and biopsy
tissues had been obtained from breast cancer patients 20-30 min afer
surgery from patients who did not receive preoperative treatment
following the appropriate ethical procedure. Tissue samples were snap
frozen in liquid nitrogen prior to transport to the Molecular Biology
lab at the Institute of Endemic Disease. Te samples were stored in a
-80°C freezer. A fraction of each sample had been stored as archival
material to be used for future needs.
Sample preparation
Tissue protein extraction had been accomplished at the Department
of Molecular Biology, Institute of Endemic Disease, University of
Khartoum, Sudan. Tissue Samples were homogenized by freezing with
liquid nitrogen and hand grinding with a cooled mortar and pestle to
a fne powder. An adequate amount of sample was suspended in lysis
bufer (9 M Urea, 4% CHAPS, 100 Mm DTT, 1% Pharmalyte pH 3-10,
and Protease inhibitor cocktail). Te suspension was vortexed for 30
seconds to facilitate protein solubilization, incubated for 1 hour at room
temperature, and centrifuged (16000 g, 30 minutes). Te supernatant
(crude extract) was transferred to a clean tube, stored at 20°C.