A unified sample preparation protocol for proteomic and genomic profiling of cervical swabs to identify biomarkers for cervical cancer screening.
Cervical cancer screening is ideally suited for the development of biomarkers due to the ease of tissue acquisition and the well-established histological transitions. Furthermore, cell and biologic fluid obtained from cervix samples undergo specific molecular changes that can be profiled. However, the ideal manner and techniques for preparing cervical samples remains to be determined. To address this critical issue a patient screening protein and nucleic acid collection protocol was established. RNAlater was used to collect the samples followed by proteomic methods to identify proteins that were differentially expressed in normal cervical epithelial versus cervical cancer cells. Three hundred ninety spots were identified via 2-D DIGE that were expressed at either higher or lower levels (>three-fold) in cervical cancer samples. These proteomic results were compared to genes in a cDNA microarray analysis of microdissected neoplastic cervical specimens to identify overlapping patterns of expression. The most frequent pathways represented by the combined dataset were: cell cycle: G2/M DNA damage checkpoint regulation; aryl hydrocarbon receptor signaling; p53 signaling; cell cycle: G1/S checkpoint regulation; and the ER stress pathway. HNRPA2B1 was identified as a biomarker candidate with increased expression in cancer compared to normal cervix and validated by Western blot.
Division of Gynecologic Oncology, Department of Obstetrics and Gynecology, Washington University School of Medicine and Alvin J. Siteman Cancer Center, St. Louis, MO, USA; Department of Genetics, Washington University School of Medicine and Alvin J. Siteman Cancer Center, St. Louis, MO, USA. raderj@wustl.edu.
Janet S
JS
Rader
James P
JP
Malone
Julia
J
Gross
Petra
P
Gilmore
Rebecca A
RA
Brooks
Loan
L
Nguyen
Dan L
DL
Crimmins
Sheng
S
Feng
Jason D
JD
Wright
Nicholas
N
Taylor
Israel
I
Zighelboim
Margo C
MC
Funk
Phyllis C
PC
Huettner
Jack H
JH
Ladenson
David
D
Gius
R Reid
RR
Townsend