Activin-A, myostatin and interleukin-6 in cancer associated cachexia

Cachexia is a muscle wasting condition associated with multiple different chronic illnesses, such as cancer, diabetes and AIDS. In cancer, approximately 80% of patients with advanced disease have symptoms of muscle wasting, and around 25% of cancer mortality concerns cachexia. Elevated serum levels...

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Main Author: Härkönen, Jouni
Other Authors: Matemaattis-luonnontieteellinen tiedekunta, Faculty of Sciences, Bio- ja ympäristötieteiden laitos, Department of Biological and Environmental Science, University of Jyväskylä, Jyväskylän yliopisto
Format: Master's thesis
Language:eng
Published: 2017
Subjects:
Online Access: https://jyx.jyu.fi/handle/123456789/54939
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author Härkönen, Jouni
author2 Matemaattis-luonnontieteellinen tiedekunta Faculty of Sciences Bio- ja ympäristötieteiden laitos Department of Biological and Environmental Science University of Jyväskylä Jyväskylän yliopisto
author_facet Härkönen, Jouni Matemaattis-luonnontieteellinen tiedekunta Faculty of Sciences Bio- ja ympäristötieteiden laitos Department of Biological and Environmental Science University of Jyväskylä Jyväskylän yliopisto Härkönen, Jouni Matemaattis-luonnontieteellinen tiedekunta Faculty of Sciences Bio- ja ympäristötieteiden laitos Department of Biological and Environmental Science University of Jyväskylä Jyväskylän yliopisto
author_sort Härkönen, Jouni
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description Cachexia is a muscle wasting condition associated with multiple different chronic illnesses, such as cancer, diabetes and AIDS. In cancer, approximately 80% of patients with advanced disease have symptoms of muscle wasting, and around 25% of cancer mortality concerns cachexia. Elevated serum levels of different cytokines and TGF-β protein family members, such as Interleukin-6, Myostatin and Activin-A, have been observed in cachetic patients and test animals. However, the mechanistic role and the relative contribution of these molecules to muscle loss in the syndrome have not yet been fully elucidated. In this thesis, the gene-expression levels of Activin-A, Myostatin and Interleukin-6 was assessed with Reverse-Transcriptase quantitative PCR from the tumor and muscle tissue of cachetic C26-tumor-bearing mice to clarify whether they arise from the tumor or are muscle derived. Additionally, transfection of CAGA-luciferase plasmid construct was optimized for the C2C12 murine myoblast cell-line to be utilized in research concerning TGF-β mediated cancer-associated cachexia in vitro. The function of the construct was tested by administration of exogenous Activin-A, with and without its inhibitor, into the C2C12 growth media. This thesis showed a marked increase in the expression levels of Activin-A (inhibin-βA) and IL-6 mRNA in cachexia inducing tumors (C26) when compared to tumors not inducing cachexia (Lewis-Lung-Cancer) (P < 0.001). Additionally, the results show decrease in muscle-derived Activin-A (inhibin-βA) in the cachetic groups (P < 0.05). These results imply that Activin-A and Interleukin-6 are strongly induced in the tumors that produced cachexia in vivo. This work also presents a successful, optimized plasmid transfection protocol for the notoriously transfection resistant C2C12-myoblast cell line. The presented in vitro –method can partially replace animal tests in related settings, when studying the mechanistic effects of cachexia-inducing TGF-β proteins on muscle tissue.
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In cancer, approximately 80% of patients with advanced disease have symptoms of muscle wasting, and around 25% of cancer mortality concerns cachexia. Elevated serum levels of different cytokines and TGF-\u03b2 protein family members, such as Interleukin-6, Myostatin and Activin-A, have been observed in cachetic patients and test animals. However, the mechanistic role and the relative contribution of these molecules to muscle loss in the syndrome have not yet been fully elucidated. In this thesis, the gene-expression levels of Activin-A, Myostatin and Interleukin-6 was assessed with Reverse-Transcriptase quantitative PCR from the tumor and muscle tissue of cachetic C26-tumor-bearing mice to clarify whether they arise from the tumor or are muscle derived. Additionally, transfection of CAGA-luciferase plasmid construct was optimized for the C2C12 murine myoblast cell-line to be utilized in research concerning TGF-\u03b2 mediated cancer-associated cachexia in vitro. 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spellingShingle Härkönen, Jouni Activin-A, myostatin and interleukin-6 in cancer associated cachexia kakeksia aktiviini-a myostatiini Cachexia Myostatin Activin-A Interleukin-6 C2C12 Solu- ja molekyylibiologia Cell and molecular biology 4013 syöpätaudit proteiinit interleukiinit
title Activin-A, myostatin and interleukin-6 in cancer associated cachexia
title_full Activin-A, myostatin and interleukin-6 in cancer associated cachexia
title_fullStr Activin-A, myostatin and interleukin-6 in cancer associated cachexia Activin-A, myostatin and interleukin-6 in cancer associated cachexia
title_full_unstemmed Activin-A, myostatin and interleukin-6 in cancer associated cachexia Activin-A, myostatin and interleukin-6 in cancer associated cachexia
title_short Activin-A, myostatin and interleukin-6 in cancer associated cachexia
title_sort activin a myostatin and interleukin 6 in cancer associated cachexia
title_txtP Activin-A, myostatin and interleukin-6 in cancer associated cachexia
topic kakeksia aktiviini-a myostatiini Cachexia Myostatin Activin-A Interleukin-6 C2C12 Solu- ja molekyylibiologia Cell and molecular biology 4013 syöpätaudit proteiinit interleukiinit
topic_facet 4013 Activin-A C2C12 Cachexia Cell and molecular biology Interleukin-6 Myostatin Solu- ja molekyylibiologia aktiviini-a interleukiinit kakeksia myostatiini proteiinit syöpätaudit
url https://jyx.jyu.fi/handle/123456789/54939 http://www.urn.fi/URN:NBN:fi:jyu-201707203334
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