Mol Biol Rep. 2012 Dec;39(12):10179-86.

Histone deacetylase inhibitor BML-210 induces growth inhibition and apoptosis and regulates HDAC and DAPC complex expression levels in cervical cancer cells.

Borutinskaite VV, Magnusson KE, Navakauskiene R.

Department of Molecular Cell Biology, Institute of Biochemistry, Vilnius University, 08662 Vilnius, Lithuania. veronika.borutinskaite@bchi.vu.lt

 

ABSTRACT:

Histone deacetylase inhibitors (HDACIs) represent a new class of targeted anti-cancer agents and different other diseases, like muscular disorders. A number of studies have shown that extracellular signal-activated kinases can target chromatin-modifying complexes directly and regulate their function. The molecular connection between the dystrophin-associated protein complex (DAPC) and chromatin has been described, by showing that NO signaling regulates HDAC2 activity and influences gene expression in different cell types.

In present study, we investigated HDACs changes in HeLa cells undergoing growth inhibition and apoptosis, caused by HDACI BML-210 and retinoic acid (ATRA). Cell cycle analysis indicated that HeLa cell treatment with 20 mikroM and 30 mikroM concentration of BML-210 increased the proportion of cells in G0/G1 phase, and caused accumulation in subG1, indicating that the cells are undergoing apoptosis. We determined down-regulation of HDAC 1-5 and 7 after treatment with BML-210. Also, we demonstrated expression of different isoforms of alpha-dystrobrevin (alpha-DB) and other components of DAPC such as syntrophin, dystrophin, beta-dystrobrevin and NOS in HeLa cells after treatments. We determined changes in protein expression level of dystrophin, NOS1, alpha- and beta-DB and in subcellular localization of alpha-DB after treatments with BML-210 and ATRA.

In conclusion, these results suggest that HDACI BML-210 can inhibit cell growth and induce apoptosis in cervical cancer cells, what correlates with down-regulation of HDAC class I and II and changes in the dystrophin-associated protein complex expression levels. This can be important for identifying target proteins in DAPC signaling to HDACs, as a target of pharmacological intervention for treatment of muscular dystrophies and other diseases.

PMID: 23007576

 

SUPPLEMENT:

The present study was designed to investigate effects of the HDACI BML-210 on the HDACs and DAPC expression during inhibition of proliferation and induction of apoptosis in HeLa cells treated with BML-210.

We showed that 20 mikroM or 30 mikroM doses of BML-210 induce growth inhibitory effect (up to 70-90 %), cell cycle arrest in G0/G1 phase and cytotoxicity through the pathway of apoptosis (up to 25 %).

BML-210, as HDAC inhibitor, probably might directly deacetylate histones or other target proteins such as transcription factors and enzymes involved in the determination of malignant cell behavior, which changes their activity, subcellular localization and interaction partners. We determined in this study that BML-210 down-regulates HDAC class I and II gene and protein expression and can be promising in cervical cancer treatment.

A number of studies have shown that extracellular signal-activated kinases can target chromatin-modifying complexes directly and regulate their function. It is important to identify target proteins in DAPC signaling to HDACs, as a target of pharmacological intervention for treatment of muscular dystrophies and other diseases. In this study, we conclude that BML-210 treatment affect DAPC protein expression level and alpha-DB subcellular localization (Fig. 1). The NOS1 up-regulation after 20 and 30 mikroM BML-210 treatments correlate with increased HeLa cell number in subG1 phase, which indicates apoptotic cells.

In conclusion, our results can be important for future looking for a target of pharmacological intervention for treatment of muscular dystrophies and other diseases.

 

Acknowledgements:

This research was supported by the Swedish Institute (Visby Program No. 00879/2009-2010), the Swedish Research Council, Lithuanian State Science and Studies Foundation (No.V-12/2009) and EU Marie Curie Programme Fellowship for V.V. Borutinskaite.

Contact:

Department of Molecular Cell Biology, Institute of Biochemistry, Vilnius University, LT-08662 Vilnius, Lithuania. Voice: +37052729327, fax: +37052729196, e-mail: veronika.borutinskaite@bchi.vu.lt; ruta.navakauskiene@bchi.vu.lt

Veronika, Borutinskaite-pic1Fig 1. HDACI BML-210 effects on NB4 cells.

Fig.2Figure 2: Members of the Proteins and epigenomics research group at the Department of Molecular Cell Biology, Institute of Biochemistry, Vilnius University.

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