Heba Shawer

Abstract

Epithelial ovarian cancer (EOC) is the most common type of ovarian tumors. The biomarkers, which are being used for EOC screening, have low sensitivity and specificity leading to late diagnosis and high mortality rate. Thus, identification of effective biomarkers for early diagnosis of ovarian cancer has become a high priority in research. Here we aim to address this problem by studying the expression of a potential molecular marker, miR-590-3p, in EOC. We examined the mRNA steady-state levels of various acknowledged biomarkers including cancer antigen-125 (CA125) which is being used for EOC screening, c-reactive protein (CRP) which expression was correlated with EOC stage and paired homeobox 2 gene (PAX2) whose expression is correlated with the histological grade to characterize the histo-pathological features of the specimens at the molecular level. We examined the expression of miR-590-3p in EOC patients’ serum and EOC tissues, using real-time PCR. The levels of circulating miR-590-3p showed to be significantly elevated in 84.6% of the EOC patients’ serum, with 76.92% sensitivity and 85.7% specificity at its optimal cutoff value. Tissue miR-590-3p showed to be upregulated in EOC tissues compared to normal ovarian tissues and highly correlated with high-grade poorly differentiated EOC. The adoption of the epithelial-mesenchymal transition biomarkers was enriched in EOC with high miR-590-3p levels. The potential downstream target genes, SOX2, LEF1 and PAX2, were predicted using miRanda and Targetscan in silco tools and their expression profiles were in silco examined using Oncomine data-mining platform and StarBase Pan-cancer analysis. We examined the levels of SOX2, LEF1 and PAX2 at the RNA and the protein levels via semi-quantitative PCR and western blot, respectively. Among the investigated potential target genes, only PAX2 showed negative correlation with the levels of miR-590-3p in EOC tissues, suggesting that miR-590-3p could acquire its role in EOC carcinogenesis through regulation of cellular differentiation in EOC.

Department

Biotechnology Program

Degree Name

MS in Biotechnology

Graduation Date

2-1-2015

Submission Date

January 2016

First Advisor

Amleh, Asma

Committee Member 1

Siam, Rania

Committee Member 2

Abou Aisha, Khalid

Extent

84 p.

Document Type

Master's Thesis

Rights

The author retains all rights with regard to copyright. The author certifies that written permission from the owner(s) of third-party copyrighted matter included in the thesis, dissertation, paper, or record of study has been obtained. The author further certifies that IRB approval has been obtained for this thesis, or that IRB approval is not necessary for this thesis. Insofar as this thesis, dissertation, paper, or record of study is an educational record as defined in the Family Educational Rights and Privacy Act (FERPA) (20 USC 1232g), the author has granted consent to disclosure of it to anyone who requests a copy.

Institutional Review Board (IRB) Approval

Approval has been obtained for this item

Comments

My deepest gratitude is to my advisor, Dr. Asma Amleh for her support, guidance, patience and encouragement. I hope that one day I would become as good to my students as Dr. Asma Amleh has been to me, and I deeply appreciate her belief in me. I would like to acknowledge Dr. Basel Refky who provided and collected the specimens that were examined in this study. My extended appreciation is to Dr. Khaled Abou Aisha who opened his lab to me to perform the real-time PCR experiments. I am also thankful to the members of Dr. Amleh’s research group. Particularly, Aya El Serw, who performed the semi-quantitative PCR experiments in this work. I am grateful to Aya for her support, care and for bearing with all my ups and downs. I would like to thank Eman El Zeneini and Mai Omar. I would like to thank my father, Mahmoud Shawer and my mother, Nagwa El-Hennawi for supporting me through my graduate studies. I am thankful to my husband and my brothers for their support and encouragement and my son, Adam Amal for inspiring me. Finally, I would also like to thank the AUC for financially supporting this work.

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