MicroRNA-3162-5p-mediated crosstalk between Kallikrein family members including prostate-specific antigen in prostate cancer

Australian Prostate Cancer BioResource, Farhana Mating, Varinder Jeet, Srilakshmi Srinivasan, Alexandre S. Cristino, Janaththani Panchadsaram, Judith A. Clements, Jyotsna Batra

Research output: Contribution to journalArticleResearchpeer-review

Abstract

BACKGROUND: MicroRNAs mediate biological processes through preferential binding to the 3' untranslated region (3' UTR) of target genes. Studies have shown their association with prostate cancer (PCa) risk through single-nucleotide polymorphisms (SNPs), known as miRSNPs. In a European cohort, 22 PCa risk-associated miRSNPs have been identified. The most significant miRSNP in the 3' UTR of Kallikrein-related peptidase 3 (KLK3) created a binding site for miR-3162-5p. Here we investigated the miR-3162-5p-KLK interaction and the clinical implication of miR-3162-5p in PCa.

METHODS: We tested the role of miR-3162-5p in PCa etiology using IncuCyte live-cell imaging and anchorage-independent growth assays. The effect of miR-3162-5p on KLK and androgen receptor (AR) expression was measured by RT-quantitative (q) PCR and target pulldown assays. KLK3 proteolytic activity was determined by DELFIA (R) immunoassay. Mass spectrometry identified pathways affected by miR-3162-5p. miR-3162-5p expression was measured in clinical samples using RT-qPCR.

RESULTS: miR-3162-5p affected proliferation, migration, and colony formation of LNCaP cells by regulating the expression of KLK2-4 and AR by direct targeting. KLK3 protein expression was regulated by miR-3162-5p consistent with lower KLK3 proteolytic activity observed in LNCaP-conditioned media. KLK/AR pulldown and mass spectrometry analysis showed a potential role of miR-3162-5p in metabolic pathways via KLK/AR and additional targets. Increased miR-3162-5p expression was observed in prostate tumor tissues with higher Gleason grade.

CONCLUSIONS: Our study provides an insight into possible involvement of miR-3162-5p in PCa etiology by targeting KLKs and AR. It highlights clinical utility of miR-3162-5p and its interactive axis as a new class of biomarkers and therapeutic targets for PCa.

LanguageEnglish
Pages771-780
Number of pages10
JournalClinical Chemistry
Volume65
Issue number6
DOIs
Publication statusPublished - Jun 2019
Externally publishedYes

Cite this

Australian Prostate Cancer BioResource, Mating, F., Jeet, V., Srinivasan, S., Cristino, A. S., Panchadsaram, J., ... Batra, J. (2019). MicroRNA-3162-5p-mediated crosstalk between Kallikrein family members including prostate-specific antigen in prostate cancer. Clinical Chemistry, 65(6), 771-780. https://doi.org/10.1373/clinchem.2018.295824
Australian Prostate Cancer BioResource ; Mating, Farhana ; Jeet, Varinder ; Srinivasan, Srilakshmi ; Cristino, Alexandre S. ; Panchadsaram, Janaththani ; Clements, Judith A. ; Batra, Jyotsna. / MicroRNA-3162-5p-mediated crosstalk between Kallikrein family members including prostate-specific antigen in prostate cancer. In: Clinical Chemistry. 2019 ; Vol. 65, No. 6. pp. 771-780.
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title = "MicroRNA-3162-5p-mediated crosstalk between Kallikrein family members including prostate-specific antigen in prostate cancer",
abstract = "BACKGROUND: MicroRNAs mediate biological processes through preferential binding to the 3' untranslated region (3' UTR) of target genes. Studies have shown their association with prostate cancer (PCa) risk through single-nucleotide polymorphisms (SNPs), known as miRSNPs. In a European cohort, 22 PCa risk-associated miRSNPs have been identified. The most significant miRSNP in the 3' UTR of Kallikrein-related peptidase 3 (KLK3) created a binding site for miR-3162-5p. Here we investigated the miR-3162-5p-KLK interaction and the clinical implication of miR-3162-5p in PCa.METHODS: We tested the role of miR-3162-5p in PCa etiology using IncuCyte live-cell imaging and anchorage-independent growth assays. The effect of miR-3162-5p on KLK and androgen receptor (AR) expression was measured by RT-quantitative (q) PCR and target pulldown assays. KLK3 proteolytic activity was determined by DELFIA (R) immunoassay. Mass spectrometry identified pathways affected by miR-3162-5p. miR-3162-5p expression was measured in clinical samples using RT-qPCR.RESULTS: miR-3162-5p affected proliferation, migration, and colony formation of LNCaP cells by regulating the expression of KLK2-4 and AR by direct targeting. KLK3 protein expression was regulated by miR-3162-5p consistent with lower KLK3 proteolytic activity observed in LNCaP-conditioned media. KLK/AR pulldown and mass spectrometry analysis showed a potential role of miR-3162-5p in metabolic pathways via KLK/AR and additional targets. Increased miR-3162-5p expression was observed in prostate tumor tissues with higher Gleason grade.CONCLUSIONS: Our study provides an insight into possible involvement of miR-3162-5p in PCa etiology by targeting KLKs and AR. It highlights clinical utility of miR-3162-5p and its interactive axis as a new class of biomarkers and therapeutic targets for PCa.",
author = "{Australian Prostate Cancer BioResource} and Farhana Mating and Varinder Jeet and Srilakshmi Srinivasan and Cristino, {Alexandre S.} and Janaththani Panchadsaram and Clements, {Judith A.} and Jyotsna Batra",
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Australian Prostate Cancer BioResource, Mating, F, Jeet, V, Srinivasan, S, Cristino, AS, Panchadsaram, J, Clements, JA & Batra, J 2019, 'MicroRNA-3162-5p-mediated crosstalk between Kallikrein family members including prostate-specific antigen in prostate cancer', Clinical Chemistry, vol. 65, no. 6, pp. 771-780. https://doi.org/10.1373/clinchem.2018.295824

MicroRNA-3162-5p-mediated crosstalk between Kallikrein family members including prostate-specific antigen in prostate cancer. / Australian Prostate Cancer BioResource; Mating, Farhana; Jeet, Varinder; Srinivasan, Srilakshmi; Cristino, Alexandre S.; Panchadsaram, Janaththani; Clements, Judith A.; Batra, Jyotsna.

In: Clinical Chemistry, Vol. 65, No. 6, 06.2019, p. 771-780.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - MicroRNA-3162-5p-mediated crosstalk between Kallikrein family members including prostate-specific antigen in prostate cancer

AU - Australian Prostate Cancer BioResource

AU - Mating, Farhana

AU - Jeet, Varinder

AU - Srinivasan, Srilakshmi

AU - Cristino, Alexandre S.

AU - Panchadsaram, Janaththani

AU - Clements, Judith A.

AU - Batra, Jyotsna

PY - 2019/6

Y1 - 2019/6

N2 - BACKGROUND: MicroRNAs mediate biological processes through preferential binding to the 3' untranslated region (3' UTR) of target genes. Studies have shown their association with prostate cancer (PCa) risk through single-nucleotide polymorphisms (SNPs), known as miRSNPs. In a European cohort, 22 PCa risk-associated miRSNPs have been identified. The most significant miRSNP in the 3' UTR of Kallikrein-related peptidase 3 (KLK3) created a binding site for miR-3162-5p. Here we investigated the miR-3162-5p-KLK interaction and the clinical implication of miR-3162-5p in PCa.METHODS: We tested the role of miR-3162-5p in PCa etiology using IncuCyte live-cell imaging and anchorage-independent growth assays. The effect of miR-3162-5p on KLK and androgen receptor (AR) expression was measured by RT-quantitative (q) PCR and target pulldown assays. KLK3 proteolytic activity was determined by DELFIA (R) immunoassay. Mass spectrometry identified pathways affected by miR-3162-5p. miR-3162-5p expression was measured in clinical samples using RT-qPCR.RESULTS: miR-3162-5p affected proliferation, migration, and colony formation of LNCaP cells by regulating the expression of KLK2-4 and AR by direct targeting. KLK3 protein expression was regulated by miR-3162-5p consistent with lower KLK3 proteolytic activity observed in LNCaP-conditioned media. KLK/AR pulldown and mass spectrometry analysis showed a potential role of miR-3162-5p in metabolic pathways via KLK/AR and additional targets. Increased miR-3162-5p expression was observed in prostate tumor tissues with higher Gleason grade.CONCLUSIONS: Our study provides an insight into possible involvement of miR-3162-5p in PCa etiology by targeting KLKs and AR. It highlights clinical utility of miR-3162-5p and its interactive axis as a new class of biomarkers and therapeutic targets for PCa.

AB - BACKGROUND: MicroRNAs mediate biological processes through preferential binding to the 3' untranslated region (3' UTR) of target genes. Studies have shown their association with prostate cancer (PCa) risk through single-nucleotide polymorphisms (SNPs), known as miRSNPs. In a European cohort, 22 PCa risk-associated miRSNPs have been identified. The most significant miRSNP in the 3' UTR of Kallikrein-related peptidase 3 (KLK3) created a binding site for miR-3162-5p. Here we investigated the miR-3162-5p-KLK interaction and the clinical implication of miR-3162-5p in PCa.METHODS: We tested the role of miR-3162-5p in PCa etiology using IncuCyte live-cell imaging and anchorage-independent growth assays. The effect of miR-3162-5p on KLK and androgen receptor (AR) expression was measured by RT-quantitative (q) PCR and target pulldown assays. KLK3 proteolytic activity was determined by DELFIA (R) immunoassay. Mass spectrometry identified pathways affected by miR-3162-5p. miR-3162-5p expression was measured in clinical samples using RT-qPCR.RESULTS: miR-3162-5p affected proliferation, migration, and colony formation of LNCaP cells by regulating the expression of KLK2-4 and AR by direct targeting. KLK3 protein expression was regulated by miR-3162-5p consistent with lower KLK3 proteolytic activity observed in LNCaP-conditioned media. KLK/AR pulldown and mass spectrometry analysis showed a potential role of miR-3162-5p in metabolic pathways via KLK/AR and additional targets. Increased miR-3162-5p expression was observed in prostate tumor tissues with higher Gleason grade.CONCLUSIONS: Our study provides an insight into possible involvement of miR-3162-5p in PCa etiology by targeting KLKs and AR. It highlights clinical utility of miR-3162-5p and its interactive axis as a new class of biomarkers and therapeutic targets for PCa.

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U2 - 10.1373/clinchem.2018.295824

DO - 10.1373/clinchem.2018.295824

M3 - Article

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EP - 780

JO - Clinical Chemistry

T2 - Clinical Chemistry

JF - Clinical Chemistry

SN - 0009-9147

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