The effect of Ephedra foeminea on human bone osteosarcoma U2OS cell viability and migration

Abstract

Although advancement has been made in the development of cancer treatments, contemporary treatments still present significant challenges such as low effectiveness and adverse side effects. There is thus a critical need to continuously develop new and more effective drugs against cancer. Herbal plants serve as a potential source for a wide variety of complex compounds with probable anticancer activity. E. foeminea is an herb whose use in the Middle East recently gained popularity as a remedy for cancer. There is however minimal empirical evidence regarding the anticancer effects of E. foeminea. In this study, the effect of E.foeminea ethyl acetate, ethanol and water extracts on morphology, viability, migratory ability and gene expression of U2OS osteosarcoma cells was examined. U2OS viability, migratory ability and the steady-state mRNA levels of genes involved in these processes were respectively studied using MTT assay, wound healing assay and reverse transcriptase PCR (RT-PCR). Results showed that all tested extracts significantly reduced U2OS percentage viability in a manner dependent on both dose and time with varying potencies; the least half maximal inhibitory concentration (IC50) recorded was that of the water extract after 48h incubation (30.761±1.4 μg/ml) followed by the ethyl acetate extract after 72h incubation (80.35±1.233 μg/ml) and finally the ethanol extract after 48h incubation (97.499±1.188 μg/ml). Ethanol extract significantly reduced U2OS percentage wound closure while both ethanol and water extracts significantly reduced the steady-state mRNA expression of Beta-catenin and its downstream targets, Twist1 and RUNX2, which are critical in promoting both proliferation and cell migration in osteosarcoma. These results suggest that E. foeminea decreases U2OS cell viability and migration by modulating the expression of key genes involved in regulating these processes.

Department

Biotechnology Program

Degree Name

MS in Biotechnology

Graduation Date

2-1-2019

Submission Date

January 2019

First Advisor

Amleh, Asma

Committee Member 1

Amleh, Asma

Committee Member 2

Abdellatif, Ahmed

Extent

74 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

Not necessary for this item

Comments

I would like to thank Dr. Asma Amleh for her mentorship and guidance throughout my graduate studies. I am grateful for her selfless dedication to making this project a success. I thank all professors in the Biotechnology Master program at AUC for their relentless endeavor in making me a better scientist and researcher; most notably I acknowledge Dr. Walid Fouad for teaching how to think critically. I am grateful to both Dr. Rami Arafeh and Mr. Ahmed El Hosseiny who were instrumental in respectively contributing towards the processing of E. foeminea crude extracts used in this study and in silico analysis of U2OS gene expression data. Many thanks go out to Dr. Asma Amleh research team (Myret Ghabriel, Mennatallah Elfar, Sheri Saleeb, Nancy Hassanein, Noha Saad, Mennatallah Ghoraba, Khaled Abdel Raouf, Rowan Bahaa, Omnia Abdelraheem, Alaa Farag and Naela Adel Saleh) for sharing their knowledge and support all through the course of this project. My appreciation goes out to my dear friends (Logayn Tarek, Ogwang Joel and Mugwanya Muziri) for their tremendous support throughout the ups and downs of grad school. Finally, I thank The American University in Cairo (AUC) for providing the African Graduate Fellowship which has facilitated my study at AUC in addition to providing the research grant without which this study would have been impossible.

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