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MicroRNAs: New Biomarkers for Diagnosis, Prognosis, Therapy Prediction and Therapeutic Tools for Breast Cancer.

Bertoli G, Cava C, Castiglioni I - Theranostics (2015)

Bottom Line: Based on the results obtained in the last decade, some miRNAs are emerging as biomarkers of BC for diagnosis (i.e., miR-9, miR-10b, and miR-17-5p), prognosis (i.e., miR-148a and miR-335), and prediction of therapeutic outcomes (i.e., miR-30c, miR-187, and miR-339-5p) and have important roles in the control of BC hallmark functions such as invasion, metastasis, proliferation, resting death, apoptosis, and genomic instability.In particular, circulating multiple miRNA profiles are showing better diagnostic and prognostic performance as well as better sensitivity than individual miRNAs in BC.New miRNA-based drugs are also promising therapy for BC (e.g., miR-9, miR-21, miR34a, miR145, and miR150), and other miRNAs are showing a fundamental role in modulation of the response to other non-miRNA treatments, being able to increase their efficacy (e.g., miR-21, miR34a, miR195, miR200c, and miR203 in combination with chemotherapy).

View Article: PubMed Central - PubMed

Affiliation: Institute of Molecular Bioimaging and Physiology (IBFM), National Research Council (CNR), Milan, Italy.

ABSTRACT
Dysregulation of microRNAs (miRNAs) is involved in the initiation and progression of several human cancers, including breast cancer (BC), as strong evidence has been found that miRNAs can act as oncogenes or tumor suppressor genes. This review presents the state of the art on the role of miRNAs in the diagnosis, prognosis, and therapy of BC. Based on the results obtained in the last decade, some miRNAs are emerging as biomarkers of BC for diagnosis (i.e., miR-9, miR-10b, and miR-17-5p), prognosis (i.e., miR-148a and miR-335), and prediction of therapeutic outcomes (i.e., miR-30c, miR-187, and miR-339-5p) and have important roles in the control of BC hallmark functions such as invasion, metastasis, proliferation, resting death, apoptosis, and genomic instability. Other miRNAs are of interest as new, easily accessible, affordable, non-invasive tools for the personalized management of patients with BC because they are circulating in body fluids (e.g., miR-155 and miR-210). In particular, circulating multiple miRNA profiles are showing better diagnostic and prognostic performance as well as better sensitivity than individual miRNAs in BC. New miRNA-based drugs are also promising therapy for BC (e.g., miR-9, miR-21, miR34a, miR145, and miR150), and other miRNAs are showing a fundamental role in modulation of the response to other non-miRNA treatments, being able to increase their efficacy (e.g., miR-21, miR34a, miR195, miR200c, and miR203 in combination with chemotherapy).

No MeSH data available.


Related in: MedlinePlus

In vitro validation of miRNA-target direct interaction. Cultured cell lines are transfected with a reporter vector containing firefly (FIR) luciferase gene and the 3′ UTR of the gene of interest (GOI). The level of expression of FIR luciferase is measured in a luminometric assay. Cells are then exposed to the mimic miRNA, which is supposed to enter within the cell and to interact with the 3′ UTR of the GOI. If no interaction between miRNA and the 3′ UTR of GOI happens (a), we could observe no alteration in the level of expression of luciferase, thus no alteration in the emitted chemoluminescence, as FIR gene produced an active, luminescent protein. The complete interaction between the miRNA and the 3′ UTR of the GOI (b) leads to reduced FIR luciferase expression, with a decrease of luminescence levels. Other luminescent genes, such as Renilla (REN) luciferase, are usually used as reference genes for luminescence normalization.
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Figure 2: In vitro validation of miRNA-target direct interaction. Cultured cell lines are transfected with a reporter vector containing firefly (FIR) luciferase gene and the 3′ UTR of the gene of interest (GOI). The level of expression of FIR luciferase is measured in a luminometric assay. Cells are then exposed to the mimic miRNA, which is supposed to enter within the cell and to interact with the 3′ UTR of the GOI. If no interaction between miRNA and the 3′ UTR of GOI happens (a), we could observe no alteration in the level of expression of luciferase, thus no alteration in the emitted chemoluminescence, as FIR gene produced an active, luminescent protein. The complete interaction between the miRNA and the 3′ UTR of the GOI (b) leads to reduced FIR luciferase expression, with a decrease of luminescence levels. Other luminescent genes, such as Renilla (REN) luciferase, are usually used as reference genes for luminescence normalization.

Mentions: Reporter assays are commonly used to study gene expression coupled with other cellular events, such as receptor activity or intracellular signal transduction of protein-protein interactions. To analyze direct miRNA-mRNA interactions, the firefly luciferase-based assay is widely used because the reporter activity is available immediately upon translation, the assay is very rapid and sensitive, and no background luminescence is found in the host cells (Figure 2). To be used as a reporter assay for validation of the interaction of a miRNA with the 3′ UTR of a gene of interest (GOI), the luciferase-based assay needs cloning of the 3′ UTR of the GOI, where the miRNA-recognized sequence is supposed to be present, downstream of the luciferase gene in the reporter vector (Figure 2). The cells are then transfected with this construct in the presence or absence of the miRNA mimic oligonucleotide. If the miRNA is able to recognize the seed in the 3′ UTR of the GOI, the level of luciferase expression is decreased, thus causing a diminished bioluminescence emission (Figure 2B); on the other hand, if the miRNA does not interact with the 3′ UTR, the emission of light is unaffected (Figure 2A). The disadvantages of this type of reporter assays are that they are laborious, expensive, sensitive only for the 3′ UTR chosen for cloning, and difficult to use for transfection 62, 63.


MicroRNAs: New Biomarkers for Diagnosis, Prognosis, Therapy Prediction and Therapeutic Tools for Breast Cancer.

Bertoli G, Cava C, Castiglioni I - Theranostics (2015)

In vitro validation of miRNA-target direct interaction. Cultured cell lines are transfected with a reporter vector containing firefly (FIR) luciferase gene and the 3′ UTR of the gene of interest (GOI). The level of expression of FIR luciferase is measured in a luminometric assay. Cells are then exposed to the mimic miRNA, which is supposed to enter within the cell and to interact with the 3′ UTR of the GOI. If no interaction between miRNA and the 3′ UTR of GOI happens (a), we could observe no alteration in the level of expression of luciferase, thus no alteration in the emitted chemoluminescence, as FIR gene produced an active, luminescent protein. The complete interaction between the miRNA and the 3′ UTR of the GOI (b) leads to reduced FIR luciferase expression, with a decrease of luminescence levels. Other luminescent genes, such as Renilla (REN) luciferase, are usually used as reference genes for luminescence normalization.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4508501&req=5

Figure 2: In vitro validation of miRNA-target direct interaction. Cultured cell lines are transfected with a reporter vector containing firefly (FIR) luciferase gene and the 3′ UTR of the gene of interest (GOI). The level of expression of FIR luciferase is measured in a luminometric assay. Cells are then exposed to the mimic miRNA, which is supposed to enter within the cell and to interact with the 3′ UTR of the GOI. If no interaction between miRNA and the 3′ UTR of GOI happens (a), we could observe no alteration in the level of expression of luciferase, thus no alteration in the emitted chemoluminescence, as FIR gene produced an active, luminescent protein. The complete interaction between the miRNA and the 3′ UTR of the GOI (b) leads to reduced FIR luciferase expression, with a decrease of luminescence levels. Other luminescent genes, such as Renilla (REN) luciferase, are usually used as reference genes for luminescence normalization.
Mentions: Reporter assays are commonly used to study gene expression coupled with other cellular events, such as receptor activity or intracellular signal transduction of protein-protein interactions. To analyze direct miRNA-mRNA interactions, the firefly luciferase-based assay is widely used because the reporter activity is available immediately upon translation, the assay is very rapid and sensitive, and no background luminescence is found in the host cells (Figure 2). To be used as a reporter assay for validation of the interaction of a miRNA with the 3′ UTR of a gene of interest (GOI), the luciferase-based assay needs cloning of the 3′ UTR of the GOI, where the miRNA-recognized sequence is supposed to be present, downstream of the luciferase gene in the reporter vector (Figure 2). The cells are then transfected with this construct in the presence or absence of the miRNA mimic oligonucleotide. If the miRNA is able to recognize the seed in the 3′ UTR of the GOI, the level of luciferase expression is decreased, thus causing a diminished bioluminescence emission (Figure 2B); on the other hand, if the miRNA does not interact with the 3′ UTR, the emission of light is unaffected (Figure 2A). The disadvantages of this type of reporter assays are that they are laborious, expensive, sensitive only for the 3′ UTR chosen for cloning, and difficult to use for transfection 62, 63.

Bottom Line: Based on the results obtained in the last decade, some miRNAs are emerging as biomarkers of BC for diagnosis (i.e., miR-9, miR-10b, and miR-17-5p), prognosis (i.e., miR-148a and miR-335), and prediction of therapeutic outcomes (i.e., miR-30c, miR-187, and miR-339-5p) and have important roles in the control of BC hallmark functions such as invasion, metastasis, proliferation, resting death, apoptosis, and genomic instability.In particular, circulating multiple miRNA profiles are showing better diagnostic and prognostic performance as well as better sensitivity than individual miRNAs in BC.New miRNA-based drugs are also promising therapy for BC (e.g., miR-9, miR-21, miR34a, miR145, and miR150), and other miRNAs are showing a fundamental role in modulation of the response to other non-miRNA treatments, being able to increase their efficacy (e.g., miR-21, miR34a, miR195, miR200c, and miR203 in combination with chemotherapy).

View Article: PubMed Central - PubMed

Affiliation: Institute of Molecular Bioimaging and Physiology (IBFM), National Research Council (CNR), Milan, Italy.

ABSTRACT
Dysregulation of microRNAs (miRNAs) is involved in the initiation and progression of several human cancers, including breast cancer (BC), as strong evidence has been found that miRNAs can act as oncogenes or tumor suppressor genes. This review presents the state of the art on the role of miRNAs in the diagnosis, prognosis, and therapy of BC. Based on the results obtained in the last decade, some miRNAs are emerging as biomarkers of BC for diagnosis (i.e., miR-9, miR-10b, and miR-17-5p), prognosis (i.e., miR-148a and miR-335), and prediction of therapeutic outcomes (i.e., miR-30c, miR-187, and miR-339-5p) and have important roles in the control of BC hallmark functions such as invasion, metastasis, proliferation, resting death, apoptosis, and genomic instability. Other miRNAs are of interest as new, easily accessible, affordable, non-invasive tools for the personalized management of patients with BC because they are circulating in body fluids (e.g., miR-155 and miR-210). In particular, circulating multiple miRNA profiles are showing better diagnostic and prognostic performance as well as better sensitivity than individual miRNAs in BC. New miRNA-based drugs are also promising therapy for BC (e.g., miR-9, miR-21, miR34a, miR145, and miR150), and other miRNAs are showing a fundamental role in modulation of the response to other non-miRNA treatments, being able to increase their efficacy (e.g., miR-21, miR34a, miR195, miR200c, and miR203 in combination with chemotherapy).

No MeSH data available.


Related in: MedlinePlus