Abstract 5056: PIM-1 kinase regulates vitamin D receptor signaling in human renal carcinoma cells

Abstract

Abstract Dysregulation of vitamin D signaling and metabolism enzymes is a frequent change in many cancers and may confound approaches to vitamin -D based therapies. The mechanisms by which aberrant vitamin D signaling and metabolism lead to resistance to vitamin D action are poorly understood. The kidney plays a critical role in vitamin D synthesis and metabolism; in renal cell carcinoma (RCC) cells vitamin D metabolism is dysregulated. Recently, we discovered that an oncogenic protein kinase PIM-1 is involved in vitamin D-induced expression of 24-hydroxylase, a key catalytic enzyme of vitamin D encoded by CYP24A1 gene, in bladder and prostate cancer. To study the impact of PIM1 on vitamin D signaling and vitamin D-mediated anti-tumor activity in RCC, we analyzed 4 human renal cell carcinoma cell lines (Caki-1, ACHN, A498 and 786-O) and 25 samples of human RCC tumor tissue and 13 benign kidney tissues (including 10 matched tumor and benign tissue samples) by qRT-PCR, immunohistochemistry (IHC), Western blot analysis, MTT and clonogenic assays. VDR mRNA and protein were detected in all RCC cell lines. IHC showed that VDR was strongly expressed in the normal kidney tissues, but decreased in RCC tissues. As expected, VDR expression was significantly increased by treatment with 1,25-dihydroxyvitamin D3 (1,25D3) in RCC cell lines. MTT and clonogenic assays showed that 1,25D3 inhibited RCC cell growth (p<0.05). We also noted a significant increase of CYP24A1 expression induced by vitamin D3 (p<0.0001); and effect which would be expected to promote the catabolism of 1,25D3. qRT-PCR and Western blot analysis demonstrated that PIM-1 is expressed in RCC cells. PIM-1 inhibition by siRNA or PIM-1 inhibitor JP_11646 significantly reduced 1,25D3-induced CYP24A1 expression (p<0.05). Therefore, we hypothesized that targeting PIM-1 kinase activity with a PIM-1 inhibitor may be an effective strategy to reduce 1,25D3 metabolism, thereby enhancing vitamin D-mediated anti-tumor activity. Inhibition of PIM1 kinase activity by JP_11646 reduced CYP24A1 expression at the transcriptional level in RCC cells. We further evaluated the efficacy of the PIM1 inhibitor alone or in combination with 1,25D3. Inhibition of PIM1 kinase activity by JP_11646 reduced cell growth in RCC cell lines in a dose-dependent manner as measured by MTT and clonogenic assays. Furthermore, inhibition of PIM1 kinase activity enhances 1,25D3-mediated inhibitory effect on cell growth in RCC cells (p<0.05). This study indicates that PIM-1 is involved in the regulation of vitamin D signaling in RCC. Inhibition of PIM kinase activity combined with vitamin D may be an attractive new therapeutic strategy for RCC. Citation Format: Wei Luo, Yingyu Ma, Carmen Baldino, Justin Caserta, Swathi Ramakrishnan, Pili Roberto, Candace Johnson, Donald Trump. PIM-1 kinase regulates vitamin D receptor signaling in human renal carcinoma cells. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 5056. doi:10.1158/1538-7445.AM2015-5056