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Nuclear localization of LDL receptor-related protein 1B in mammary gland carcinogenesis

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A Commentary to this article was published on 15 March 2019

Abstract

LRP1B intracellular domain is released and transported to the nucleus; however, pathological consequences of this nuclear transport are largely unclear. We aimed to unravel the pathobiological significance of nuclear localization of LRP1B intracellular domain in mammary gland carcinogenesis. Immunohistochemical staining using antibodies for LRP1B intracellular domain was performed to determine LRP1B expression in 92 invasive ductal breast carcinomas. LRP1B immunoreactivity was detected in the surface membrane and cytoplasm of 60 of 92 invasive ductal carcinomas and in the nucleus of 15 of 92 carcinomas. Nuclear LRP1B was significantly associated with poor patient prognosis, particularly luminal A type breast cancer, where it was significantly related to nodal metastasis. Doxycycline-dependent nuclear expression of LRP1B intracellular domain was established in cultured breast cancer cells. Enforced nuclear expression significantly increased Matrigel invasion activity in MCF-7 and T47D luminal A breast cancer cells. Moreover, enforced nuclear expression of LRP1B intracellular domain facilitated MCF-7 cells growth in mammary fat pad of nude mice, which was supplemented with estrogen. Comprehensive microarray-based analysis demonstrated that nuclear expression of LRP1B intracellular domain significantly increased long non-coding RNA nuclear paraspeckle assembly transcript 1 (NEAT1) expression, which facilitates breast cancer invasion with poor prognosis. Nuclear-localized LRP1B intracellular domain promoted breast cancer progression with poor prognosis, possibly through the NEAT1 pathway. Nuclear transport of LRP1B intracellular domain could be a therapeutic target for breast cancer patients.

Key messages

  • Nuclear LRP1B was significantly associated with poor patient prognosis.

  • Nuclear LRP1B increased Matrigel invasion activity of breast cancer cells.

  • Nuclear expression of LRP1B intracellular domain increased NEAT1 expression.

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Funding

This study was supported by grants from the Ministry of Education of Japan (grant nos. KAKEN 15K08361, 15K19051, and 17K15642).

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Authors and Affiliations

  1. Department of Surgical Oncology, Gifu University, Gifu University Graduate School of Medicine, Gifu, Japan

    Yoshimi Asano, Yoshinori Iwata, Manabu Futamura & Kazuhiro Yoshida

  2. Department of Pathology and Translational Research, Gifu University Graduate School of Medicine, Gifu, Japan

    Tamotsu Takeuchi, Hiroshi Okubo, Chiemi Saigo & Yusuke Kito

  3. Department of Breast and Molecular Oncology, Gifu University Graduate School of Medicine, Gifu, Japan

    Manabu Futamura

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  1. Yoshimi Asano
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Correspondence to Tamotsu Takeuchi.

Ethics declarations

Paraffin-embedded tissues surgically resected from patients with breast cancer were retrospectively used after diagnosis. The need for written informed consent was waived by the Institutional Review Board of the Gifu University Graduate School of Medicine. Instead, the Institutional Review Board requested us to inform the patients that they could refuse the use of their tissue specimens for this study if they did not want to participate in the present study. The experimental protocol to obtain the antibodies was approved by the Animal Care Committee of Gifu University. The present study was conducted in accordance with the ethical standards of the Helsinki Declaration in 1975, after approval of the Institutional Review Board of the Gifu University Graduate School of Medicine (specific approval number 25–81).

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Suppl File 1

Doxycycline treatment did not affect endogenous LRP1B mRNA transcript expression in MCF-7 and T47D cells. The value for the groups (n = 3) was calculated as the fold-change relative to the mean value for the control non-treated group (control set to 1.0). Standard deviations were then computed for the triplicate sets. Three target genes and fold changes are presented. Student’s t test was performed to determine the significant differences. P = 0.93 (24 h), P = 0.86 (48 h), P = 0.79 (72 h), and P = 0.76 (96 h) in MCF-7 cells. P = 0.15 (24 h), P = 0.80 (48 h), P = 0.66 (72 h), and P = 0.34 (96 h) in T47D cells. (PNG 149 kb)

High resolution image (TIF 56 kb)

Suppl File 2

Doxycycline-controlled expression of LRP1B intracellular domain increased MUC1 expression but did not increased nuclear MUC1-CTD in MCF-7 cells. Expression of LRP1B intracellular domain was induced by doxycycline at 100 ng/mL (indicated as Tet-on), and without doxycycline (indicated as Tet-off). Note the marked diminishment of MUC1 immunoreactivity in MUC1-targeted siRNA treated cells. (PNG 15 kb)

High resolution image (TIF 808 kb)

Suppl File 3

A: Both MUC1 targeting siRNAs markedly impaired expression of MUC1 and MUC1-CTD protein. B: Down-regulation of MUC1 did not alter the expression of MALAT1 or NEAT1. C: Down-regulation MUC1 did not alter the Matrigel invasion activity in LRP1B intracellular domain increased MCF-7 cells (PNG 624 kb)

High resolution image (TIF 178 kb)

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Asano, Y., Takeuchi, T., Okubo, H. et al. Nuclear localization of LDL receptor-related protein 1B in mammary gland carcinogenesis. J Mol Med 97, 257–268 (2019). https://doi.org/10.1007/s00109-018-01732-2

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  • DOI: https://doi.org/10.1007/s00109-018-01732-2

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