Anticancer Drug Development : Evaluation of Anti-Proliferative Activity of the Seeds of Dicerocaryum Seneciodes on Leukemia T Cell Line.

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Evaluation of the phytochemical composition and the anti-proliferative potential of seed extract of Dicerocaryum senecioides on Jurkat T cell lines By Humbulani Ronald Tshilongamulenzhe A Dissertation submitted to the Faculty of Science and Agriculture, Department of Biochemistry, Microbiology and Biotechnology, School of Molecular and Life Sciences in accordance with the requirements of the University of Limpopo for honours for the degree Bachelor of Science in Biochemistry

UNIVERSITY OF LIMPOPO SOVENGA 0727 SOUTH AFRICA

Supervisor: Prof L.J. Mampuru

NOVEMBER 2011

DECLARATION I declare that the work presented in this dissertation submitted to the University of Limpopo for the degree of Honours in Biochemistry, is my own in design and execution and all information contained herein has been accordingly acknowledged.

Tshilongamulenzhe Humbulani Ronald Student no. 200810649

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DEDICATION I dedicate this work to my parents, Mashudu and Agnes Tshilongamulenzhe, my sister (Mukovhe), my aunt (Regina), my uncles (Titus and Mphelekedzeni), my grandmothers (Johanna and Muofhe) and my pastors (Mr and Mrs Tovhakale) who contributed to my success.

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ACKNOWLEDGEMENTS Firstly, I would like to give thanks to the Almighty God for strengthening me from the beginning of this work until its completion. For it was by His Grace which made it possible for me to pull through this endeavour. I wish to express my heartfelt appreciation to my supervisor, Prof L.J. Mampuru for his guidance, support, suggestions, and encouragement throughout the course of this research. Thank you for trusting and teaching me on how to be self-governing. Special thanks are extended also to different individuals most especially Dr K.C Lucas who strengthened and gave me the guidance before I began with the research. To Dr Mbazima, Dr Mokgotho, Dr T.M Matsibatlela, Ms Pilane and Ms Ntini, thank you for teaching me how to handle the cells. I remembered Dr Mbazima and Dr Mokgotho saying I should handle them like babies. Thank you very much. I also wish to express a word of appreciation to Mr M. Mokhubedi for availing and fixing some laboratory equipment, Ms A. Mothiba for cleaning the laboratory and Ms T. Mabokela for cleaning laboratory glassware. To my entire family, thank you for support and the love you gave me throughout my academic years. I also appreciate Honours students for encouragement and the spirit of unity that they showed me. Thank you guys! I also wish to express my sincere appreciation to the National Research Foundation (NRF) for the grant they provided for this research.

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TABLE OF CONTENTS Page Declaration Dedication Acknowledgements Table of contents List of abbreviations List of figures Abstract CHAPTER 1: INTRODUCTION 1. Introduction 1.1 What is cancer 1.2 Causes of cancer 1.3 Carcinogenesis 1.4 Leukaemia 1.5 Cell line used 1.6 Cell death 1.6.1 Apoptosis 1.6.2 Necrosis 1 1-2 2 3 3-4 4 4-5 5 5-6 v i ii iii iv vii viii ix

1.7 Cancer treatment 1.7.1 Chemotherapy 1.7.2 Radiation 1.7.3 Surgery 1.8 Medicinal plants 1.8.1 Phytochemicals on cancer 1.8.2 Antifungal peptides 1.8.3 Medicinal plant used in the study 1.9 Rationale of the study 1.10 Aim and objectives of the study 1.10.1 Aim 1.10.2 Objectives CHAPTER 2: MATERIAL AND METHODS 2.1 Materials 2.2 Methods 2.2.1 Preparation of plant extracts 2.2.2 Thin layer chromatography 2.2.2.1 Phytochemical analysis 2.2.2.2 Assay for antioxidant activity

6 7 7 8 8-9 9 9-10 10-11 11 12 12 12

13 13 13-14 14 14 14-15 vi

2.2.3 Cell cultures and treatment 2.2.4 Assay for antiproliferative activity toward Jurkat T cell line CHAPTER 3: RESULTS 3.1 Phytochemical composition 3.2 Antioxidant potential 3.3 Antiproliferative activity CHAPTER 4: DISCUSSION AND CONCLUSION CHAPTER 5: REFERENCES

15 15

16-17 17 18 19-21 22-24

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LIST OF ABBREVIATIONS ATCC BAW CEF CO2 DPPH EMW FBS IC50 PSN RPMI-1640 TLC American Tissue Culture Collection Butanol: Acetic acid: Water...
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