Tumor Biology

, Volume 36, Issue 6, pp 4417–4426 | Cite as

Protein kinase D2 silencing reduced motility of doxorubicin-resistant MCF7 cells

Research Article

Abstract

Success of chemotherapy is generally impaired by multidrug resistance, intrinsic resistance, or acquired resistance to functionally and structurally irrelevant drugs. Multidrug resistance emerges via distinct mechanisms: increased drug export, decreased drug internalization, dysfunctional apoptotic machinery, increased DNA damage repair, altered cell cycle regulation, and increased drug detoxification. Several reports demonstrated that multidrug resistance is a multifaceted problem such that multidrug resistance correlates with increased aggressiveness and metastatic potential. Here, we tested the involvement of protein kinase D2, a serine/threonine kinase that was previously implicated in proliferation, drug resistance, and motility in doxorubicin-resistant MCF7 (MCF7/DOX) cell line, which served as an in vitro model for drug resistance and invasiveness. We showed that basal level activity of protein kinase D2 (PKD2) was higher in MCF7/DOX cells than parental MCF7 cells. To elucidate the roles of PKD2 MCF7/DOX, PKD2 expression was reduced via small interfering RNA (siRNA)-mediated knockdown. Results showed that acquired resistance of MCF7/DOX to doxorubicin was not affected by PKD2 silencing, while motility of MCF7/DOX cells was reduced. The results implied that PKD2 silencing might inhibit migration of MCF7/DOX cells without affecting chemoresistance significantly.

Keywords

Multidrug resistance Breast cancer Migration Protein kinase D2 

Notes

Acknowledgments

We thank Assoc. Dr. Özlem Darcansoy İşeri and Assoc. Dr. Meltem Demirel Kars for developing MCF7/DOX subline and Çağrı Urfalı Mamatoğlu for critical reading. We greatly acknowledged Soner Yildiz and Assoc. Prof. Dr. Mayda Gürsel for their contributions in flow cytometry. This work was supported by TUBITAK 1002 Short Term R&D Funding Program (Grant ID: 112T714) and METU Research Fund (Grant ID: BAP-07-02-2012-101-22).

Conflicts of interest

None

Supplementary material

13277_2015_3081_Fig8_ESM.gif (203 kb)
Supplementary Fig. 1

PKD2 knockdown increases the migration of parental MCF7 cell line. MCF7 cells were transfected for 72 h. Cells are collected and seeded at a density of 100 000 cells in 200 μl 0.5 % FBS medium per transwell insert. Cells are allowed to migrate toward 10 % FBS medium for 48 h. (A) RT-qPCR analysis demonstrated that PKD2 levels remained downregulated at fifth day. (B) Migrating cells on transwell membrane were fixed, stained and counted. Representative images of membranes were taken at 20X magnification. (GIF 202 kb)

13277_2015_3081_MOESM1_ESM.tif (912 kb)
High-resolution image (TIFF 911 kb)

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Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2015

Authors and Affiliations

  1. 1.Department of Biological SciencesMiddle East Technical UniversityAnkaraTurkey

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