Lentiviral shRNA screen of human kinases identifies PLK1 as a potential therapeutic target for osteosarcoma

Zhenfeng Duan, Diana Ji, Edward J. Weinstein, Xianzhe Liu, Michiro Susa, Edwin Choy, Cao Yang, Henry Mankin, Francis J. Hornicek

Research output: Contribution to journalArticle

44 Citations (Scopus)

Abstract

We describe an optimized systematic screen of known kinases using osteosarcoma cell lines (KHOS and U-2OS) and a lentiviral-based short hairpin RNA (shRNA) human kinase library. CellTiter 96®AQueous One Solution Cell Proliferation Assay was used to measure cell growth and survival. We identified several kinases, including human polo-like kinase (PLK1), which inhibit cell growth and induce apoptosis in osteosarcoma cells when knocked down. cDNA rescue and synthetic siRNA assays confirm that the observed phenotypic changes result from the loss of PLK1 gene expression.Furthermore, a small molecule inhibitor to PLK1 inhibited osteosarcoma cell growth and induced apoptosis. Western blot analysis confirmed that PLK1 is highly expressed and activated in several osteosarcoma cell lines as well as in resected tumor samples. Immunohistochemistry analysis showed that patients with high PLK1 tumor expression levels correlated with significantly shorter survival than patients with lower levels of tumor PLK1 expression. These results demonstrate the capability and feasibility of a high-throughput screen with a large collection of lentiviral kinases and its effectiveness in identifying potential drug targets. The development of more potent inhibitors that target PLK1 may open doors to a new range of anti-cancer strategies in osteosarcoma.

Original languageEnglish
Pages (from-to)220-229
Number of pages10
JournalCancer Letters
Volume293
Issue number2
DOIs
Publication statusPublished - 2010 Jul
Externally publishedYes

Fingerprint

Osteosarcoma
Small Interfering RNA
Phosphotransferases
Neoplasms
Growth
Apoptosis
Therapeutics
Cell Line
Cell Survival
Complementary DNA
Western Blotting
Immunohistochemistry
Cell Proliferation
Gene Expression
Survival
Pharmaceutical Preparations

Keywords

  • Osteosarcoma
  • PLK1
  • ShRNA

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Duan, Z., Ji, D., Weinstein, E. J., Liu, X., Susa, M., Choy, E., ... Hornicek, F. J. (2010). Lentiviral shRNA screen of human kinases identifies PLK1 as a potential therapeutic target for osteosarcoma. Cancer Letters, 293(2), 220-229. https://doi.org/10.1016/j.canlet.2010.01.014

Lentiviral shRNA screen of human kinases identifies PLK1 as a potential therapeutic target for osteosarcoma. / Duan, Zhenfeng; Ji, Diana; Weinstein, Edward J.; Liu, Xianzhe; Susa, Michiro; Choy, Edwin; Yang, Cao; Mankin, Henry; Hornicek, Francis J.

In: Cancer Letters, Vol. 293, No. 2, 07.2010, p. 220-229.

Research output: Contribution to journalArticle

Duan, Z, Ji, D, Weinstein, EJ, Liu, X, Susa, M, Choy, E, Yang, C, Mankin, H & Hornicek, FJ 2010, 'Lentiviral shRNA screen of human kinases identifies PLK1 as a potential therapeutic target for osteosarcoma', Cancer Letters, vol. 293, no. 2, pp. 220-229. https://doi.org/10.1016/j.canlet.2010.01.014
Duan, Zhenfeng ; Ji, Diana ; Weinstein, Edward J. ; Liu, Xianzhe ; Susa, Michiro ; Choy, Edwin ; Yang, Cao ; Mankin, Henry ; Hornicek, Francis J. / Lentiviral shRNA screen of human kinases identifies PLK1 as a potential therapeutic target for osteosarcoma. In: Cancer Letters. 2010 ; Vol. 293, No. 2. pp. 220-229.
@article{8465b150a621492086c70001a77d573f,
title = "Lentiviral shRNA screen of human kinases identifies PLK1 as a potential therapeutic target for osteosarcoma",
abstract = "We describe an optimized systematic screen of known kinases using osteosarcoma cell lines (KHOS and U-2OS) and a lentiviral-based short hairpin RNA (shRNA) human kinase library. CellTiter 96{\circledR}AQueous One Solution Cell Proliferation Assay was used to measure cell growth and survival. We identified several kinases, including human polo-like kinase (PLK1), which inhibit cell growth and induce apoptosis in osteosarcoma cells when knocked down. cDNA rescue and synthetic siRNA assays confirm that the observed phenotypic changes result from the loss of PLK1 gene expression.Furthermore, a small molecule inhibitor to PLK1 inhibited osteosarcoma cell growth and induced apoptosis. Western blot analysis confirmed that PLK1 is highly expressed and activated in several osteosarcoma cell lines as well as in resected tumor samples. Immunohistochemistry analysis showed that patients with high PLK1 tumor expression levels correlated with significantly shorter survival than patients with lower levels of tumor PLK1 expression. These results demonstrate the capability and feasibility of a high-throughput screen with a large collection of lentiviral kinases and its effectiveness in identifying potential drug targets. The development of more potent inhibitors that target PLK1 may open doors to a new range of anti-cancer strategies in osteosarcoma.",
keywords = "Osteosarcoma, PLK1, ShRNA",
author = "Zhenfeng Duan and Diana Ji and Weinstein, {Edward J.} and Xianzhe Liu and Michiro Susa and Edwin Choy and Cao Yang and Henry Mankin and Hornicek, {Francis J.}",
year = "2010",
month = "7",
doi = "10.1016/j.canlet.2010.01.014",
language = "English",
volume = "293",
pages = "220--229",
journal = "Cancer Letters",
issn = "0304-3835",
publisher = "Elsevier Ireland Ltd",
number = "2",

}

TY - JOUR

T1 - Lentiviral shRNA screen of human kinases identifies PLK1 as a potential therapeutic target for osteosarcoma

AU - Duan, Zhenfeng

AU - Ji, Diana

AU - Weinstein, Edward J.

AU - Liu, Xianzhe

AU - Susa, Michiro

AU - Choy, Edwin

AU - Yang, Cao

AU - Mankin, Henry

AU - Hornicek, Francis J.

PY - 2010/7

Y1 - 2010/7

N2 - We describe an optimized systematic screen of known kinases using osteosarcoma cell lines (KHOS and U-2OS) and a lentiviral-based short hairpin RNA (shRNA) human kinase library. CellTiter 96®AQueous One Solution Cell Proliferation Assay was used to measure cell growth and survival. We identified several kinases, including human polo-like kinase (PLK1), which inhibit cell growth and induce apoptosis in osteosarcoma cells when knocked down. cDNA rescue and synthetic siRNA assays confirm that the observed phenotypic changes result from the loss of PLK1 gene expression.Furthermore, a small molecule inhibitor to PLK1 inhibited osteosarcoma cell growth and induced apoptosis. Western blot analysis confirmed that PLK1 is highly expressed and activated in several osteosarcoma cell lines as well as in resected tumor samples. Immunohistochemistry analysis showed that patients with high PLK1 tumor expression levels correlated with significantly shorter survival than patients with lower levels of tumor PLK1 expression. These results demonstrate the capability and feasibility of a high-throughput screen with a large collection of lentiviral kinases and its effectiveness in identifying potential drug targets. The development of more potent inhibitors that target PLK1 may open doors to a new range of anti-cancer strategies in osteosarcoma.

AB - We describe an optimized systematic screen of known kinases using osteosarcoma cell lines (KHOS and U-2OS) and a lentiviral-based short hairpin RNA (shRNA) human kinase library. CellTiter 96®AQueous One Solution Cell Proliferation Assay was used to measure cell growth and survival. We identified several kinases, including human polo-like kinase (PLK1), which inhibit cell growth and induce apoptosis in osteosarcoma cells when knocked down. cDNA rescue and synthetic siRNA assays confirm that the observed phenotypic changes result from the loss of PLK1 gene expression.Furthermore, a small molecule inhibitor to PLK1 inhibited osteosarcoma cell growth and induced apoptosis. Western blot analysis confirmed that PLK1 is highly expressed and activated in several osteosarcoma cell lines as well as in resected tumor samples. Immunohistochemistry analysis showed that patients with high PLK1 tumor expression levels correlated with significantly shorter survival than patients with lower levels of tumor PLK1 expression. These results demonstrate the capability and feasibility of a high-throughput screen with a large collection of lentiviral kinases and its effectiveness in identifying potential drug targets. The development of more potent inhibitors that target PLK1 may open doors to a new range of anti-cancer strategies in osteosarcoma.

KW - Osteosarcoma

KW - PLK1

KW - ShRNA

UR - http://www.scopus.com/inward/record.url?scp=77952518695&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=77952518695&partnerID=8YFLogxK

U2 - 10.1016/j.canlet.2010.01.014

DO - 10.1016/j.canlet.2010.01.014

M3 - Article

C2 - 20144850

AN - SCOPUS:77952518695

VL - 293

SP - 220

EP - 229

JO - Cancer Letters

JF - Cancer Letters

SN - 0304-3835

IS - 2

ER -