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Characterization of non-coding RNAs in Human cancer

Scicluna, Patrick (2012) MOBT19 20112
Degree Projects in Molecular Biology
Abstract
Abstract

Non-coding RNAs (ncRNAs) are involved in many diverse biological functions. In this thesis, my objectives were focused on the functional evaluation of two types of ncRNAs. First, the functional consequences of miR-944 regulation were evaluated in HeLa, a cervical cancer cell line. Results showed a reduced cell proliferation on miR-944 overexpression by using the WST-1 assay. In addition, 51 miR-944 targets were identified by using a recently described biochemical method. In the second part of the project, I attempted to investigate the functional
role of TDRG1, a testis-specific ncRNA known to give rise to germ cell-specific piRNAs, in Tcam2, a human testicular germ cell tumor cell line. Preliminary results showed that... (More)
Abstract

Non-coding RNAs (ncRNAs) are involved in many diverse biological functions. In this thesis, my objectives were focused on the functional evaluation of two types of ncRNAs. First, the functional consequences of miR-944 regulation were evaluated in HeLa, a cervical cancer cell line. Results showed a reduced cell proliferation on miR-944 overexpression by using the WST-1 assay. In addition, 51 miR-944 targets were identified by using a recently described biochemical method. In the second part of the project, I attempted to investigate the functional
role of TDRG1, a testis-specific ncRNA known to give rise to germ cell-specific piRNAs, in Tcam2, a human testicular germ cell tumor cell line. Preliminary results showed that increased expression of TDRG1 decreased cell proliferation in Tcam2; however, the effect was not significant. In summary, my findings revealed a novel functional role and targets of miR-944 in human cervical cancer, which may lead to a better understanding of its role in cervical cancer development and its potential use as a diagnostic biomarker.

Popular science summary:

Non-coding (nc) RNA and cancer

Recent discoveries in modern biology revealed surprising findings about the human genome. Only a very small percentage of the transcribed RNA is translated into a protein product while most RNAs form part of the newly discovered RNA types, known as ‘non-coding RNAs’ that are not translated. Non-coding RNAs can be broadly divided on the bases of size into small and long ncRNAs. Small RNAs (sRNA) were first discovered in 1993 in the worm and later found to function as regulators of gene expression by inhibiting translation of mRNA. There are different types of small RNAs that are classified by the way they inhibit mRNA translation and by how they are generated. Additionally, certain sRNAs have a highly specialized function and are generally present in a specific tissue-type. Long ncRNAs are similar to mRNA transcripts but generally do not code for a protein product. They were found to be involved in heterogeneous biological functions that ultimately determine which genes are expressed.

Scientific evidence has shown a strong link between ncRNA derangement and cancer. Indeed ncRNAs were found to be a major player determining the outcome of different oncologic diseases in different tissue types. The extensive role of ncRNAs in diverse biological processes makes this field a priority for research, in order to understand the steps leading to cancer development.
The aims of this project were to understand the function of two types of ncRNAs. The first was a sRNA, known as ‘miR-944’, associated with cervical cancer. We performed in vitro experiments to characterize its role in human cervical cancer using biochemical and flow-cytometric methods. Furthermore, we expanded this study to experimentally determine the interactions of miR-944 with mRNA transcripts by applying a state-of-the-art technique. Understanding these molecular interactions or ‘targets’ is very challenging but promising approach that can improve current cancer detecting methods. The second RNA was a long ncRNA that is specifically expressed in sexually reproductive cells. This long ncRNA is interesting since it gives rise to other small ncRNAs.

Significance of findings
Our findings indicated a decrease in cell growth of a specific type of cervical cancer cells that were overexpressing miR-944 and novel molecular targets were identified for miR-944. Furthermore, preliminary data on our long ncRNA showed that overexpression of this gene also seem to inhibit cell growth.

In brief, we concluded that miR-944 has the potential to be used as a selective tumour marker. Additionally, the novel miR-944 molecular targets revealed, broaden its functional role in human cancer. These encouraging results are another step forward towards understanding the mechanisms involved in tumor development at the molecular level.

Advisor: Ass. Prof. Weng-Onn Lui (Department of Molecular Medicine & Surgery; Karolinska Institutet)
Master´s Degree Project 60 credits in Molecular Genetics/non-coding RNAs and cancer (2012).
Department of Biology, Lund University. (Less)
Please use this url to cite or link to this publication:
author
Scicluna, Patrick
supervisor
organization
course
MOBT19 20112
year
type
H2 - Master's Degree (Two Years)
subject
language
English
id
3632864
date added to LUP
2013-04-11 11:54:31
date last changed
2013-04-11 11:54:31
@misc{3632864,
  abstract     = {Abstract

Non-coding RNAs (ncRNAs) are involved in many diverse biological functions. In this thesis, my objectives were focused on the functional evaluation of two types of ncRNAs. First, the functional consequences of miR-944 regulation were evaluated in HeLa, a cervical cancer cell line. Results showed a reduced cell proliferation on miR-944 overexpression by using the WST-1 assay. In addition, 51 miR-944 targets were identified by using a recently described biochemical method. In the second part of the project, I attempted to investigate the functional
role of TDRG1, a testis-specific ncRNA known to give rise to germ cell-specific piRNAs, in Tcam2, a human testicular germ cell tumor cell line. Preliminary results showed that increased expression of TDRG1 decreased cell proliferation in Tcam2; however, the effect was not significant. In summary, my findings revealed a novel functional role and targets of miR-944 in human cervical cancer, which may lead to a better understanding of its role in cervical cancer development and its potential use as a diagnostic biomarker.

Popular science summary:

Non-coding (nc) RNA and cancer

Recent discoveries in modern biology revealed surprising findings about the human genome. Only a very small percentage of the transcribed RNA is translated into a protein product while most RNAs form part of the newly discovered RNA types, known as ‘non-coding RNAs’ that are not translated. Non-coding RNAs can be broadly divided on the bases of size into small and long ncRNAs. Small RNAs (sRNA) were first discovered in 1993 in the worm and later found to function as regulators of gene expression by inhibiting translation of mRNA. There are different types of small RNAs that are classified by the way they inhibit mRNA translation and by how they are generated. Additionally, certain sRNAs have a highly specialized function and are generally present in a specific tissue-type. Long ncRNAs are similar to mRNA transcripts but generally do not code for a protein product. They were found to be involved in heterogeneous biological functions that ultimately determine which genes are expressed.

Scientific evidence has shown a strong link between ncRNA derangement and cancer. Indeed ncRNAs were found to be a major player determining the outcome of different oncologic diseases in different tissue types. The extensive role of ncRNAs in diverse biological processes makes this field a priority for research, in order to understand the steps leading to cancer development.
The aims of this project were to understand the function of two types of ncRNAs. The first was a sRNA, known as ‘miR-944’, associated with cervical cancer. We performed in vitro experiments to characterize its role in human cervical cancer using biochemical and flow-cytometric methods. Furthermore, we expanded this study to experimentally determine the interactions of miR-944 with mRNA transcripts by applying a state-of-the-art technique. Understanding these molecular interactions or ‘targets’ is very challenging but promising approach that can improve current cancer detecting methods. The second RNA was a long ncRNA that is specifically expressed in sexually reproductive cells. This long ncRNA is interesting since it gives rise to other small ncRNAs. 

Significance of findings
Our findings indicated a decrease in cell growth of a specific type of cervical cancer cells that were overexpressing miR-944 and novel molecular targets were identified for miR-944. Furthermore, preliminary data on our long ncRNA showed that overexpression of this gene also seem to inhibit cell growth.

In brief, we concluded that miR-944 has the potential to be used as a selective tumour marker. Additionally, the novel miR-944 molecular targets revealed, broaden its functional role in human cancer. These encouraging results are another step forward towards understanding the mechanisms involved in tumor development at the molecular level.

Advisor: Ass. Prof. Weng-Onn Lui (Department of Molecular Medicine & Surgery; Karolinska Institutet)
Master´s Degree Project 60 credits in Molecular Genetics/non-coding RNAs and cancer (2012).
Department of Biology, Lund University.},
  author       = {Scicluna, Patrick},
  language     = {eng},
  note         = {Student Paper},
  title        = {Characterization of non-coding RNAs in Human cancer},
  year         = {2012},
}