Abstract

Entacmaea quadricolor (Phylum Cnidaria, class Anthozoa) is a marine anemone found in Coral Reefs in the Red Sea. Its venom is reported to be a potential anticancer agent when tested on several cancer cell lines; such as skin cancer, and lung cancer cell lines. Yet, none of the earlier studies have characterized the extracted venom, nor tested its activity on hepatocellular carcinoma (HCC). The purpose of this study was to determine for the first time the potential cytotoxic activity of the Red Sea anemone E. quadricolor on HCC cell line. In addition to the effects of bleaching, seasonality and light exposure during storage on the venom were studied. Moreover, it was aimed to determine whether the cytotoxic activity was apoptotic or necrotic. In this study, the venom of the anemone E. quadricolor was extracted and stored under three different conditions; winter vs. summer, light vs. dark and bleached vs. unbleached. The cytotoxic activity of each venom was tested on SNU-449 HCC cells using the MTT assay. SDS-PAGE was used to differentiate between venoms based on protein composition. Moreover, Annexin-V/PI assay was used to determine the type of cytotoxic activity. The results revealed that E. quadricolor had potent cytotoxic activity against SNU-449 cells that was mediated by a necrotic pathway. The maximum activity was found during summer at the half-inhibitory concentration (IC50) of 20 µg/ ml. However, this cytotoxic activity was neutralized when the venom had been exposed to light when stored. Furthermore, cytotoxic activity was significantly decreased upon Bleaching of the anemone. A protein of 28 kDa was found in the composition of the venoms of bleached and unbleached organisms possibly identifying the cytotoxic active protein. The present results underline the findings of previous studies showing the cytotoxic activity of the sea anemone venom on cancer cell lines extending this to HCC. Furthermore, the findings are unique in showing that a bleached organism still produces toxic proteins and the venom loses its toxicity when exposed to light.

Department

Biotechnology Program

Degree Name

MS in Biotechnology

Graduation Date

6-1-2018

Submission Date

February 2018

First Advisor

Bos, Arthur R.

Committee Member 1

Amleh, Asma

Committee Member 2

Abdellatif, Ahmed

Extent

47 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

Praise is to ALLAH by Whose grace good deeds are completed I would like to show the deepest gratitude to my advisors; Dr Arthur Bos and Dr Asma Amleh for their continuous guidance and support. And to extend my sincere graduate to Dr. Ahmed Moustafa for offering his help when needed. In addition, I would like to express my gratitude to Dr Hamza El Dorry for his valuable advice and to Dr Michail kontominas at chemistry department AUC as well. Special thanks to Professor Mehmet Ozturk from the Department of Molecular Biology and Genetics, Bilkent University for kindly providing SNU-449 cell line as a gift from him. I highly appreciate Mona El Rady and Mohammed Maged for their valuable advices that helped me a lot in the practical work and thesis writing. I extremely thank Amgad Ouf and Aya Youssef for their continued support. To all my friends and colleagues, I would like to express my appreciation especially to, Menna Ghouraba, Nouran Adly, Youssef Abdo, Laila Ziko, Menna El Far, Nancy Youssef, Noha Saad, Nahla Hussien, Sara Sonbol and Myreit Ghabrial. I specially thank the most helpful lab aids; Mr Zien and Mr Mohammed. Finally, I would like to express much gratitude to the AUC for providing the grant that funded this research and for partially funding my studies at the AUC.

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