Vitamin K2 induces non-apoptotic cell death along with autophagosome formation in breast cancer cell lines

  • Shinsuke Miyazawa
  • Shota Moriya
  • Hiroko Kokuba
  • Hirotsugu Hino
  • Naoharu Takano
  • Keisuke MiyazawaEmail author
Original Article



Vitamin K2 (VK2) has been reported to induce apoptosis in many types of cancer cells including leukemia. However, there are no precise reports regarding the breast cancer cells. From the stand point of clinical implications of VK2 including chemoprevention, we investigated the effects of VK2 on breast cancer cell lines.


Breast cancer cell lines were cultured with VK2, and the cytotoxicity and cell death phenotype were examined. The HL-60 leukemia cells were used as a control for VK2-induced apoptosis.


VK2 exhibited the cytotoxic effect, especially in triple negative breast cancer cell lines, namely, MDA-MB-231 and MDA-MB-468. However, in contrast to HL-60 cells, typical features of the cells undergoing apoptosis, such as chromatin condensation, nuclear fragments, and cleavage of caspase-3 were not detected. Transmission electron microscopy exhibited an increased number of autophagosomes/autolysosomes with plasma membrane integrity. An autophagy inhibitor, 3-methyladenine, apparently attenuated VK2-induced cytotoxicity, which indicated the involvement of autophagy-dependent cell death. Interestingly, both VK2-induced non-apoptotic cell death in MDA-MB-231 cells and VK2-induced apoptosis in HL-60 cells were suppressed in the presence of reactive oxygen species (ROS) scavengers. Therefore, ROS production by VK2 seems to be located up-stream in the molecular machinery for both the types of cell death execution.


The VK2 induced non-apoptotic cell death along with autophagy, in triple negative breast cancer cell lines. Cell death phenotype induced by VK2 appears to differ among the type of cancers. This suggests the possibility of using VK2 for the breast cancer therapy.


Breast cancer Vitamin K2 Autophagy Apoptosis Reactive oxygen species 



This study was supported by funds provided through the Strategic Research Foundation at Private Universities (S1411011, 2014–2018) from the Ministry of Education, Culture, Sports, Science and Technology (MEXT) of Japan and by JSPS KAKENHI Grant Numbers JP26460478 to KM. We would like to thank Editage ( for English language editing.

Compliance with ethical standards

Conflict of interest

All authors declare that there is no conflict of interest.

Supplementary material

12282_2019_1012_MOESM1_ESM.pptx (83 kb)
Supplementary file1 (PPTX 83 kb) Supplemental Fig.1. Non-apoptotic cell death induction in breast cancer cell lines after treatment with VK2. Cells were treated with the indicated concentrations of VK2 for 24–96 h. Cellular proteins were extracted and analyzed by immunoblotting using anti-cleaved caspase-3 mAb. C-96 represents the cells cultured in control medium for 96 h. Posi-C represents HL-60 cell treated with staurosporine at 1 µM for 8 h as a positive control for apoptosis induction


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Copyright information

© The Japanese Breast Cancer Society 2019

Authors and Affiliations

  1. 1.Department of Pharmaceutical SciencesMeiji Pharmaceutical UniversityTokyoJapan
  2. 2.Department of BiochemistryTokyo Medical UniversityTokyoJapan
  3. 3.Joint Research for Basic Medical Science, Institute of Medical ScienceTokyo Medical UniversityTokyoJapan

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