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PI3K-like kinases restrain Pim gene expression in endothelial cells

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Summary

Pim kinases contribute to tumor formation and development of lymphoma, which shows enhanced DNA replication, DNA recombination and repair. Endothelial cells^(ECs) express all the three members of Pim kinase gene family. We hypothesized that DNA repair gene would regulate Pim expression in ECs. Human umbilical vein endothelial cells (HUVECs) were isolated and maintained in M199 culture medium. The cellular distribution of Pim-3 in ECs was determined by immunofluorescent staining. The siRNA fragments were synthesized and transfected by using Lipofectamine LTX. The total cellular RNA was extracted from the cells by using Trizol reagent. cDNAs were quantified by semi-quantity PCR. The effects of LY294002 and wortmannin on RNA stability in ECs were also examined. Our data showed that LY294002 and wortmannin, phosphatidylinositol 3-kinase (PI3K) and PI3K-like kinase inhibitors, increased Pim mRNA expression in ECs without altering the mRNA stability. RNA interference (RNAi) targeting DNA-dependent protein kinase catalytic subunit (DNA-PKcs) and ataxia telangiectasia mutated (ATM) increased mRNA expression of Pim-3 and Pim-1, respectively. Silencing of Akt decreased Pim-1 instead of Pm-2 and Pim-3 gene expression in ECs. But etoposide, a nucleoside analogue, which could activate DNA-PKcs and ATM, increased Pim expression in ECs. Our study indicates that the expression of Pim kinases is physiologically related to DNA-PKcs and ATM in ECs.

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Authors and Affiliations

  1. Department of Cardiology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China

    Xinwen Min  (闵新文) & Yexin Ma  (马业新)

  2. Institute of Cardiovascular Science, Dongfeng Hospital, Hubei University of Medicine, Shiyan, 442008, China

    Xinwen Min  (闵新文), Libing Zhao  (赵黎丙), Handong Yang  (杨汉东) & Peng Zhang  (张 鹏)

  3. Department of Anatomy, Hubei University of Medicine, Shiyan, 442000, China

    Jie Tang  (唐 杰) & Minghua Yu  (余明华)

  4. Department of Physiology, Hubei University of Medicine, Shiyan, 442000, China

    Yinfang Wang  (汪引芳)

Authors
  1. Xinwen Min  (闵新文)
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  2. Jie Tang  (唐 杰)
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  4. Minghua Yu  (余明华)
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  8. Yexin Ma  (马业新)
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Correspondence to Yexin Ma  (马业新).

Additional information

This project was supported by grants from the National Natural Science Foundation of China (No. 30900631) and Hubei Provincial Department of Education (No. Q20102101 and D20082406).

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Min, X., Tang, J., Wang, Y. et al. PI3K-like kinases restrain Pim gene expression in endothelial cells. J. Huazhong Univ. Sci. Technol. [Med. Sci.] 32, 17–23 (2012). https://doi.org/10.1007/s11596-012-0003-0

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  • DOI: https://doi.org/10.1007/s11596-012-0003-0

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