High-throughput screens identify microRNAs essential for HER2 positive breast cancer cell growth

Suvi-Katri Leivonen (Corresponding Author), Kristine Kleivi Sahlberg, Rami Mäkelä, Eldri Undlien Due, Olli Kallioniemi, Anne-Lise Børresen-Dale, Merja Perälä

Research output: Contribution to journalArticleScientificpeer-review

97 Citations (Scopus)

Abstract

MicroRNAs (miRNAs) are non-coding RNAs regulating gene expression post-transcriptionally. We have characterized the role of miRNAs in regulating the human epidermal growth factor receptor 2 (HER2)-pathway in breast cancer. We performed miRNA gain-of-function assays by screening two HER2 amplified cell lines (KPL-4 and JIMT-1) with a miRNA mimic library consisting of 810 human miRNAs. The levels of HER2, phospho-AKT, phospho-ERK1/2, cell proliferation (Ki67) and apoptosis (cPARP) were analyzed with reverse-phase protein arrays. Rank product analyses identified 38 miRNAs (q <0.05) as inhibitors of HER2 signaling and cell growth, the most effective being miR-491-5p, miR-634, miR-637 and miR-342-5p. We also characterized miRNAs directly targeting HER2 and identified seven novel miRNAs (miR-552, miR-541, miR-193a-5p, miR-453, miR-134, miR-498, and miR-331-3p) as direct regulators of the HER2 3'UTR. We demonstrated the clinical relevance of the miRNAs and identified miR-342-5p and miR-744* as significantly down-regulated in HER2-positive breast tumors as compared to HER2-negative tumors from two cohorts of breast cancer patients (101 and 1302 cases). miR-342-5p specifically inhibited HER2-positive cell growth, as it had no effect on the growth of HER2-negative control cells in vitro. Furthermore, higher expression of miR-342-5p was associated with better survival in both breast cancer patient cohorts. In conclusion, we have identified miRNAs which are efficient negative regulators of the HER2 pathway that may play a role in vivo during breast cancer progression. These results give mechanistic insights in HER2 regulation which may open potential new strategies towards prevention and therapeutic inhibition of HER2-positive breast cancer.
Original languageEnglish
Pages (from-to)93-104
JournalMolecular Oncology
Volume8
Issue number1
DOIs
Publication statusPublished - 2014
MoE publication typeA1 Journal article-refereed

Fingerprint

MicroRNAs
Breast Neoplasms
Growth
human ERBB2 protein
Protein Array Analysis
Untranslated RNA
3' Untranslated Regions
Libraries
Cell Proliferation
Apoptosis
Gene Expression

Keywords

  • breast cancer
  • functional screen
  • HER2
  • MicroRNA
  • reverse-phase protein array

Cite this

Leivonen, S-K., Sahlberg, K. K., Mäkelä, R., Due, E. U., Kallioniemi, O., Børresen-Dale, A-L., & Perälä, M. (2014). High-throughput screens identify microRNAs essential for HER2 positive breast cancer cell growth. Molecular Oncology, 8(1), 93-104. https://doi.org/10.1016/j.molonc.2013.10.001
Leivonen, Suvi-Katri ; Sahlberg, Kristine Kleivi ; Mäkelä, Rami ; Due, Eldri Undlien ; Kallioniemi, Olli ; Børresen-Dale, Anne-Lise ; Perälä, Merja. / High-throughput screens identify microRNAs essential for HER2 positive breast cancer cell growth. In: Molecular Oncology. 2014 ; Vol. 8, No. 1. pp. 93-104.
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Leivonen, S-K, Sahlberg, KK, Mäkelä, R, Due, EU, Kallioniemi, O, Børresen-Dale, A-L & Perälä, M 2014, 'High-throughput screens identify microRNAs essential for HER2 positive breast cancer cell growth', Molecular Oncology, vol. 8, no. 1, pp. 93-104. https://doi.org/10.1016/j.molonc.2013.10.001

High-throughput screens identify microRNAs essential for HER2 positive breast cancer cell growth. / Leivonen, Suvi-Katri (Corresponding Author); Sahlberg, Kristine Kleivi; Mäkelä, Rami; Due, Eldri Undlien; Kallioniemi, Olli; Børresen-Dale, Anne-Lise; Perälä, Merja.

In: Molecular Oncology, Vol. 8, No. 1, 2014, p. 93-104.

Research output: Contribution to journalArticleScientificpeer-review

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T1 - High-throughput screens identify microRNAs essential for HER2 positive breast cancer cell growth

AU - Leivonen, Suvi-Katri

AU - Sahlberg, Kristine Kleivi

AU - Mäkelä, Rami

AU - Due, Eldri Undlien

AU - Kallioniemi, Olli

AU - Børresen-Dale, Anne-Lise

AU - Perälä, Merja

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N2 - MicroRNAs (miRNAs) are non-coding RNAs regulating gene expression post-transcriptionally. We have characterized the role of miRNAs in regulating the human epidermal growth factor receptor 2 (HER2)-pathway in breast cancer. We performed miRNA gain-of-function assays by screening two HER2 amplified cell lines (KPL-4 and JIMT-1) with a miRNA mimic library consisting of 810 human miRNAs. The levels of HER2, phospho-AKT, phospho-ERK1/2, cell proliferation (Ki67) and apoptosis (cPARP) were analyzed with reverse-phase protein arrays. Rank product analyses identified 38 miRNAs (q <0.05) as inhibitors of HER2 signaling and cell growth, the most effective being miR-491-5p, miR-634, miR-637 and miR-342-5p. We also characterized miRNAs directly targeting HER2 and identified seven novel miRNAs (miR-552, miR-541, miR-193a-5p, miR-453, miR-134, miR-498, and miR-331-3p) as direct regulators of the HER2 3'UTR. We demonstrated the clinical relevance of the miRNAs and identified miR-342-5p and miR-744* as significantly down-regulated in HER2-positive breast tumors as compared to HER2-negative tumors from two cohorts of breast cancer patients (101 and 1302 cases). miR-342-5p specifically inhibited HER2-positive cell growth, as it had no effect on the growth of HER2-negative control cells in vitro. Furthermore, higher expression of miR-342-5p was associated with better survival in both breast cancer patient cohorts. In conclusion, we have identified miRNAs which are efficient negative regulators of the HER2 pathway that may play a role in vivo during breast cancer progression. These results give mechanistic insights in HER2 regulation which may open potential new strategies towards prevention and therapeutic inhibition of HER2-positive breast cancer.

AB - MicroRNAs (miRNAs) are non-coding RNAs regulating gene expression post-transcriptionally. We have characterized the role of miRNAs in regulating the human epidermal growth factor receptor 2 (HER2)-pathway in breast cancer. We performed miRNA gain-of-function assays by screening two HER2 amplified cell lines (KPL-4 and JIMT-1) with a miRNA mimic library consisting of 810 human miRNAs. The levels of HER2, phospho-AKT, phospho-ERK1/2, cell proliferation (Ki67) and apoptosis (cPARP) were analyzed with reverse-phase protein arrays. Rank product analyses identified 38 miRNAs (q <0.05) as inhibitors of HER2 signaling and cell growth, the most effective being miR-491-5p, miR-634, miR-637 and miR-342-5p. We also characterized miRNAs directly targeting HER2 and identified seven novel miRNAs (miR-552, miR-541, miR-193a-5p, miR-453, miR-134, miR-498, and miR-331-3p) as direct regulators of the HER2 3'UTR. We demonstrated the clinical relevance of the miRNAs and identified miR-342-5p and miR-744* as significantly down-regulated in HER2-positive breast tumors as compared to HER2-negative tumors from two cohorts of breast cancer patients (101 and 1302 cases). miR-342-5p specifically inhibited HER2-positive cell growth, as it had no effect on the growth of HER2-negative control cells in vitro. Furthermore, higher expression of miR-342-5p was associated with better survival in both breast cancer patient cohorts. In conclusion, we have identified miRNAs which are efficient negative regulators of the HER2 pathway that may play a role in vivo during breast cancer progression. These results give mechanistic insights in HER2 regulation which may open potential new strategies towards prevention and therapeutic inhibition of HER2-positive breast cancer.

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KW - functional screen

KW - HER2

KW - MicroRNA

KW - reverse-phase protein array

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SN - 1574-7891

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Leivonen S-K, Sahlberg KK, Mäkelä R, Due EU, Kallioniemi O, Børresen-Dale A-L et al. High-throughput screens identify microRNAs essential for HER2 positive breast cancer cell growth. Molecular Oncology. 2014;8(1):93-104. https://doi.org/10.1016/j.molonc.2013.10.001