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Analysis of the p53 tumour suppressor protein activation in Xenopus embryos

Analysis of the p53 tumour suppressor protein activation in Xenopus embryos

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P53 is a tumor suppressor protein whose activation may play a role in the transition from radio-resistance to radio-sensitivity in early Xenopus embryos. Specifically we compared the radiation response in normal and p53 depleted embryos. It can be concluded that the transition from radio-resistance to radio-sensitivity may represent endogenous changes in the degree of p53 activation in the Xenopus embryo.
P53 is a tumor suppressor protein whose activation may play a role in the transition from radio-resistance to radio-sensitivity in early Xenopus embryos. Specifically we compared the radiation response in normal and p53 depleted embryos. It can be concluded that the transition from radio-resistance to radio-sensitivity may represent endogenous changes in the degree of p53 activation in the Xenopus embryo.

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Published by: Undergraduate Awards on Sep 01, 2012
Copyright:Attribution Non-commercial

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10/27/2013

 
Analysis of the p53 tumour suppressor proteinactivation in
 Xenopus
embryos
Lynne O’SheaPHAR 40020: Pharmacology Research Project
This thesis is submitted in partial fulfillment of a B.Sc. honours degree in pharmacology
Supervisor: Dr. Carmel Hensey
 
Title PageSummary..….…………………………………………………...………..……iIntroduction………………………………………………………….....……..1
 Xenopus
general development……………………………………….....……...1Radiation Response…………………………………………………….………3Transcription factor p53, what it is and what it does…………………….…….5Morpholinos, anti-sense oligonucleotides…………………………….………..8
Experimental Procudures……………………………………………………9
Materials……………………………………………………………………….9Embryological methods………………………………………………………..9Morpholino’s and Micro-injections…………………………………………...10Embryonic death and survival curves…………………………………………11
 Xenopus
Extracts…………………………………………………………….. 12Separation of Proteins in extract…………………………………………...….12Western blotting…………………………………………………………….....13
Results………………………………………………………………………...15
 p53 present in
 Xenopus
Embryos……………………………………………..15
γ 
-irradiation and
 Xenopus
Embryos………………………………………..…15The effect of p53 morpholino (MO) injection………………………………...18
Discussion…………………………………………………………………......25
Concluding Remarks…………………………………………………………..29
Acknowledgements………………………………………………….….…….29Abbreviations…………………………………………………………………30References………………………………………………………….…………31
 
Summary:
The aim of this research project was to analyse the role of p53 activation in the transitionfrom radio-resistance to radio-sensitivity in early
 Xenopus
embryos. Specifically wecompared the radiation response in normal and p53 depleted embryos.The ability of the tumor suppressor protein p53 to trigger apoptosis is an essential defenseagainst cancer. Half of all human tumours express wild type p53 and therefore activationof p53 is thought to offer a new therapeutic strategy. p53 is activated by DNA damage,this can be caused by exposure to radiation. Two types of event can be triggered by theactivation of p53: cell cycle arrest or apoptosis. Embryos undergo a period of rapid celldivision during which radiation exposure does not cause cell cycle arrest i.e. they areradio-resistant. As the cell cycle develops and becomes more complex the cells of theembryos become more sensitive to radiation, here cells respond by activating a cell death pathway.The results obtained showed some evidence that p53 depletion in
 Xenopus
embryoscauses the radiation response to be activated at a later stage than in normal embryos. p53is stably expressed in the embryo, however the acquisition of radiosensitivity correspondsto a time when the subcellular localisation of p53 changes from cytoplasmic to nuclear.It can be concluded that the transition from radio-resistant to radio-sensitive mayrepresent endogenous changes in the degree of p53 activation in the
 Xenopus
embryo.Further understanding of the endogenous mechanisms of p53 activation is important for  pursuing p53 as a therapeutic target.
i

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