Genome-wide functional analysis of microRNAs in prostate cancer

Päivi Östling, Suvi-Katri Leivonen, Anna Aakula, Pekka Kohonen, Rami Mäkelä, Petri Saviranta, Merja Perälä, Olli Kallioniemi

Research output: Contribution to conferenceConference articleScientific

Abstract

MicroRNAs (miRNAs) are a novel class of negative gene regulators that have a major modulatory impact on the protein expression levels. They base-pair primarily to the 3'-untranslated region (3'UTR) of their target mRNAs leading to translational repression or transcript degradation. Thus far, studies on differential expression of miRNAs in prostate cancer have been published but functional proof of their impact in prostate cancer pathogenesis remains largely unexplored. In this study, we aimed to systematically identify cancer-associated miRNAs and their functional significance by combining data from overexpression phenotypes of individual miRNAs, with expression profiles of miRNA and mRNA. We screened two large-scale miRNA libraries one from Ambion (pre-miR V2; with 319 molecules) and one from Dharmacon (Mimic v.10.1; 820 molecules). The miRNAs were reverse-transfected in 384-well plates that were incubated for 72 h. Cell proliferation was measured by CellTiter-Glor, and apoptosis by Apo-ONEr (Promega). The screens were done in two biological replicates with several malignant and non-malignant prostate cell lines (LNCaP, LAPC4, MDA-PCa-2b, 22Rv1, CWR-1R, RWPE-1, and EP156T). In the secondary validation and follow-up screens, further phenotypic analyses were performed by utilizing the protein lysate microarray (LMA) technology with antibodies against markers for proliferation (Ki67) and apoptosis (cleaved PARP). Anti-proliferative effects of mir-15a and mir-16 were evident in LNCaP and LAPC4 cells both in the primary screens and in LMA. In addition to these miRNAs, with previously described links to prostate cancer, several novel ones were identified. However, detailed molecular mechanisms and the signalling pathways targeted by these miRNAs require further exploration. For example, we are combining predicted protein target data from these miRNAs with simultaneously measured expression profiles from mRNAs and miRNAs in order to narrow the search to the most important target proteins. In summary, we are combining various functional methods to identify miRNA genes and their mechanisms of action on prostate cancer development and progression.
Original languageEnglish
Publication statusPublished - 2009
MoE publication typeNot Eligible
EventProstate Cancer Translational Research in Europe - Amsterdam, Netherlands
Duration: 22 Jun 200923 Jun 2009

Conference

ConferenceProstate Cancer Translational Research in Europe
CountryNetherlands
CityAmsterdam
Period22/06/0923/06/09

Fingerprint

MicroRNAs
Prostatic Neoplasms
Genome
Messenger RNA
Apoptosis
Protein Array Analysis
Proteins
3' Untranslated Regions
Regulator Genes
Base Pairing
Prostate
Cell Proliferation
Technology
Phenotype
Cell Line

Cite this

Östling, P., Leivonen, S-K., Aakula, A., Kohonen, P., Mäkelä, R., Saviranta, P., ... Kallioniemi, O. (2009). Genome-wide functional analysis of microRNAs in prostate cancer. Paper presented at Prostate Cancer Translational Research in Europe, Amsterdam, Netherlands.
Östling, Päivi ; Leivonen, Suvi-Katri ; Aakula, Anna ; Kohonen, Pekka ; Mäkelä, Rami ; Saviranta, Petri ; Perälä, Merja ; Kallioniemi, Olli. / Genome-wide functional analysis of microRNAs in prostate cancer. Paper presented at Prostate Cancer Translational Research in Europe, Amsterdam, Netherlands.
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Östling, P, Leivonen, S-K, Aakula, A, Kohonen, P, Mäkelä, R, Saviranta, P, Perälä, M & Kallioniemi, O 2009, 'Genome-wide functional analysis of microRNAs in prostate cancer' Paper presented at Prostate Cancer Translational Research in Europe, Amsterdam, Netherlands, 22/06/09 - 23/06/09, .

Genome-wide functional analysis of microRNAs in prostate cancer. / Östling, Päivi; Leivonen, Suvi-Katri; Aakula, Anna; Kohonen, Pekka; Mäkelä, Rami; Saviranta, Petri; Perälä, Merja; Kallioniemi, Olli.

2009. Paper presented at Prostate Cancer Translational Research in Europe, Amsterdam, Netherlands.

Research output: Contribution to conferenceConference articleScientific

TY - CONF

T1 - Genome-wide functional analysis of microRNAs in prostate cancer

AU - Östling, Päivi

AU - Leivonen, Suvi-Katri

AU - Aakula, Anna

AU - Kohonen, Pekka

AU - Mäkelä, Rami

AU - Saviranta, Petri

AU - Perälä, Merja

AU - Kallioniemi, Olli

PY - 2009

Y1 - 2009

N2 - MicroRNAs (miRNAs) are a novel class of negative gene regulators that have a major modulatory impact on the protein expression levels. They base-pair primarily to the 3'-untranslated region (3'UTR) of their target mRNAs leading to translational repression or transcript degradation. Thus far, studies on differential expression of miRNAs in prostate cancer have been published but functional proof of their impact in prostate cancer pathogenesis remains largely unexplored. In this study, we aimed to systematically identify cancer-associated miRNAs and their functional significance by combining data from overexpression phenotypes of individual miRNAs, with expression profiles of miRNA and mRNA. We screened two large-scale miRNA libraries one from Ambion (pre-miR V2; with 319 molecules) and one from Dharmacon (Mimic v.10.1; 820 molecules). The miRNAs were reverse-transfected in 384-well plates that were incubated for 72 h. Cell proliferation was measured by CellTiter-Glor, and apoptosis by Apo-ONEr (Promega). The screens were done in two biological replicates with several malignant and non-malignant prostate cell lines (LNCaP, LAPC4, MDA-PCa-2b, 22Rv1, CWR-1R, RWPE-1, and EP156T). In the secondary validation and follow-up screens, further phenotypic analyses were performed by utilizing the protein lysate microarray (LMA) technology with antibodies against markers for proliferation (Ki67) and apoptosis (cleaved PARP). Anti-proliferative effects of mir-15a and mir-16 were evident in LNCaP and LAPC4 cells both in the primary screens and in LMA. In addition to these miRNAs, with previously described links to prostate cancer, several novel ones were identified. However, detailed molecular mechanisms and the signalling pathways targeted by these miRNAs require further exploration. For example, we are combining predicted protein target data from these miRNAs with simultaneously measured expression profiles from mRNAs and miRNAs in order to narrow the search to the most important target proteins. In summary, we are combining various functional methods to identify miRNA genes and their mechanisms of action on prostate cancer development and progression.

AB - MicroRNAs (miRNAs) are a novel class of negative gene regulators that have a major modulatory impact on the protein expression levels. They base-pair primarily to the 3'-untranslated region (3'UTR) of their target mRNAs leading to translational repression or transcript degradation. Thus far, studies on differential expression of miRNAs in prostate cancer have been published but functional proof of their impact in prostate cancer pathogenesis remains largely unexplored. In this study, we aimed to systematically identify cancer-associated miRNAs and their functional significance by combining data from overexpression phenotypes of individual miRNAs, with expression profiles of miRNA and mRNA. We screened two large-scale miRNA libraries one from Ambion (pre-miR V2; with 319 molecules) and one from Dharmacon (Mimic v.10.1; 820 molecules). The miRNAs were reverse-transfected in 384-well plates that were incubated for 72 h. Cell proliferation was measured by CellTiter-Glor, and apoptosis by Apo-ONEr (Promega). The screens were done in two biological replicates with several malignant and non-malignant prostate cell lines (LNCaP, LAPC4, MDA-PCa-2b, 22Rv1, CWR-1R, RWPE-1, and EP156T). In the secondary validation and follow-up screens, further phenotypic analyses were performed by utilizing the protein lysate microarray (LMA) technology with antibodies against markers for proliferation (Ki67) and apoptosis (cleaved PARP). Anti-proliferative effects of mir-15a and mir-16 were evident in LNCaP and LAPC4 cells both in the primary screens and in LMA. In addition to these miRNAs, with previously described links to prostate cancer, several novel ones were identified. However, detailed molecular mechanisms and the signalling pathways targeted by these miRNAs require further exploration. For example, we are combining predicted protein target data from these miRNAs with simultaneously measured expression profiles from mRNAs and miRNAs in order to narrow the search to the most important target proteins. In summary, we are combining various functional methods to identify miRNA genes and their mechanisms of action on prostate cancer development and progression.

M3 - Conference article

ER -

Östling P, Leivonen S-K, Aakula A, Kohonen P, Mäkelä R, Saviranta P et al. Genome-wide functional analysis of microRNAs in prostate cancer. 2009. Paper presented at Prostate Cancer Translational Research in Europe, Amsterdam, Netherlands.