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. 2011 Oct 20;105(10):1563–1573. doi: 10.1038/bjc.2011.426

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Copyright © 2011 Cancer Research UK

This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material.If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit https://creativecommons.org/licenses/by/4.0/.

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PIM-1 is overexpressed in RCC cell lines and inhibition of PIM kinase activity with SGI-1776 promotes apoptosis and reduced viability. (A) PIM-1 kinase expression in RCC cell lines. Immunoblotting demonstrates elevated PIM-1 levels compared with normal RPTECs. Band intensity was quantified using ImageJ software. (B) SGI-1776 reduces RCC cell viability. Five RCC cell lines and RPTEC cells were incubated with the indicated concentrations of SGI-1776 for 72 h and cell viability was measured by MTT assay. Mean±s.d., n=3. (C) SGI-1776 reduces RCC clonogenic survival. Cells were treated with indicated concentrations of SGI-1776 for 24 h, washed and incubated in fresh media for 10 days. Colonies were fixed, stained with crystal violet and quantified. Mean±s.d., n=3. ^*Indicates a significant difference compared with control. P<0.05. (D) SGI-1776 decreases Bad phosphorylation at Ser112. Cells were treated with SGI-1776 for 48 h and phospho- and total Bad levels were determined by immunoblotting. Phospho-Bad band intensity was quantified using ImageJ software. (E) SGI-1776 induces apoptosis in RCC cell lines. Cells were treated with SGI-1776 for 48 h and apoptosis was measured by PI-FACS analysis. Mean±s.d., n=3. ^*Indicates a significant difference compared with control. P<0.05.