Abstract

Ovarian cancer remains one of the leading causes of cancer-related mortality among

women, highlighting the need for novel therapeutic agents with improved selectivity and

reduced toxicity. Anticancer peptides (ACPs) have emerged as promising candidates

due to their ability to selectively target cancer cells. In this study, the anticancer activity

of a computationally designed ACP was evaluated in vitro using the SKOV-3 ovarian

cancer cell line, with HEK cells employed as a non-cancerous control.

Cell viability was assessed using the MTT assay, and cytotoxicity was further confirmed

by the trypan blue exclusion method. Apoptosis and necrosis were analyzed using

Annexin V-FITC/Propidium iodide staining. The effects of ACP treatment on cell

migration and invasion were examined using wound-healing and Transwell invasion

assays, respectively. Quantitative real-time PCR (qPCR) was performed to evaluate the

expression of genes associated with apoptosis, proliferation, and

epithelial–mesenchymal transition (EMT), with GAPDH used as a housekeeping gene.

ACP treatment resulted in a dose-dependent reduction in SKOV-3 cell viability, while

minimal cytotoxic effects were observed in HEK cells, indicating selective anticancer

activity. Trypan blue analysis confirmed increased cell death following ACP treatment.

Apoptosis analysis revealed a significant increase in early apoptotic cells in ACP-treated

SKOV-3 cells compared with untreated controls. Functional assays demonstrated a

marked inhibition of cell migration and invasion upon ACP treatment. Gene expression

analysis showed modulation of apoptosis- and proliferation-associated markers;

however, no statistically significant changes were observed in the EMT-related genes

TWIST1 and N-cadherin.

This study provides functional evidence that the computationally designed anticancer

peptide (ACP) exhibits selective anticancer activity against ovarian cancer cells in vitro,

associated with increased apoptosis and reduced migratory and invasive behaviors.

School

School of Sciences and Engineering

Department

Biotechnology Program

Degree Name

MS in Biotechnology

Graduation Date

Summer 6-1-2025

Submission Date

2-12-2025

First Advisor

Dr. Asma Amleh

Committee Member 1

Dr. Ahmed Abdellatif

Committee Member 2

Dr. Rania Hassan Mohamed

Committee Member 3

De. Andrea Kakarougkas

Extent

47 p.

Document Type

Master's Thesis

Institutional Review Board (IRB) Approval

Not necessary for this item

Disclosure of AI Use

Thesis text drafting

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