DNA Aptamers against Vaccinia-Related Kinase (VRK) 1 Block Proliferation in MCF7 Breast Cancer Cells.

Víctor M González,Rebeca Carrión-Marchante,Ana Salgado-Figueroa,M Elena Martín,Valerio Frezza,M Isabel Pérez-Morgado

doi:10.3390/ph14050473

Abstract

Vaccinia-related kinase (VRK) 1 is a serin/threonine kinase that plays an important role in DNA damage response (DDR), phosphorylating some proteins involved in this process such as 53BP1, NBS1 or H2AX, and in the cell cycle progression. In addition, VRK1 is overexpressed in many cancer types and its correlation with poor prognosis has been determined, showing VRK1 as a new therapeutic target in oncology. Using in vitro selection, high-affinity DNA aptamers to VRK1 were selected from a library of ssDNA. Selection was monitored using the enzyme-linked oligonucleotide assay (ELONA), and the selected aptamer population was cloned and sequenced. Three aptamers were selected and characterized. These aptamers recognized the protein kinase VRK1 with an affinity in the nanomolar range and showed a high sensibility. Moreover, the treatment of the MCF7 breast cell line with these aptamers resulted in a decrease in cyclin D1 levels, and an inhibition of cell cycle progression by G1 phase arrest, which induced apoptosis in cells. These results suggest that these aptamers are specific inhibitors of VRK1 that might be developed as potential drugs for the treatment of cancer.

Highlights

IntroductionThe kinase proteins Vaccinia-related kinase (VRK) (vaccinia-related kinases) are part of serin/threonin-kinases family [1]
The kinase proteins Vaccinia-related kinase (VRK) are part of serin/threonin-kinases family [1]
Some proteins involved in DNA damage response (DDR) are phosphorylated by VRK1 such as 53BP1 [5], NBS1 [6] or H2AX [7] which in its phosphorylated state acts as a platform of recruitment for other proteins involved in DDR [8]

Summary

Introduction

The kinase proteins VRKs (vaccinia-related kinases) are part of serin/threonin-kinases family [1]. Some proteins involved in DNA damage response (DDR) are phosphorylated by VRK1 such as 53BP1 [5], NBS1 [6] or H2AX [7] which in its phosphorylated state acts as a platform of recruitment for other proteins involved in DDR [8]. VRK1 forms a complex and phosphorylates p53 mediating protective responses for the cells [9,10]. All these data suggest an important role for VRK1 in the signaling of DNA damage responses [11]

Methods

Results

Conclusion