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Pinch-1 was up-regulated in leukemia BMSC and its possible effect

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Abstract

Pinch-1, a widely expressed focal adhesion protein, has been demonstrated to be up-regulated in multiple solid tumor-associated stromal cells, particularly at invasive edges. It was supposed that Pinch-1 was intimately associated with development and progression of tumors. The expression of Pinch-1 in hematopoietic microenvironment in patients with leukemia remains unclear. This study focused on the expression of Pinch-1 in bone marrow stromal cells (BMSCs) from leukemia patients and its possible effect. BMSC was isolated and cultured from bone marrow in leukemia patients and normal healthy donors. RT-PCR and Western blot analysis were performed to determine Pinch-1 mRNA and protein level in BMSC, respectively. Lentiviral vector containing Pinch-1 siRNA was constructed, and the recombinant lentivirus particle was packaged in 293 cells. Effectiveness of Pinch-1 siRNA was determined by Western blot. The proliferation, apoptosis and motility of leukemia BMSC subjected to Pinch-1 knockdown using siRNA were tested by flow cytometry, TUNEL assay and Transwell system, respectively. Pinch-1 mRNA and protein were significantly up-regulated in ALL and AML BMSC compared to normal BMSC (p < 0.01). Although there was no difference in Pinch-1 mRNA between ALL and AML BMSC, cellular levels of Pinch-1 protein in ALL BMSC were significantly higher than that in AML BMSC (p < 0.01). Overexpressed Pinch-1 was significantly reduced in leukemia BMSC transfected with Pinch-1 siRNA evidenced by Western blot. Flow cytometry analysis showed that the percentage of cells in S + G2 phases in leukemia BMSC transfected with Pinch-1 siRNA was significantly lower than control (p < 0.01). The percentage of apoptotic cells in leukemia BMSC transfected with Pinch-1 siRNA was 19.8 ± 1.0%, significantly higher than controls (p < 0.01). The number of leukemia BMSC transfected with Pinch-1 siRNA that migrated to the lower chamber after culturing for 24 h was 8.4 ± 1.1 per field, significantly lower than controls (p < 0.01). Pinch-1 mRNA and protein in leukemia BMSC were up-regulated drastically compared with BMSC from healthy donors. Leukemia BMSC displayed hypoproliferation, decreased migration and increased apoptosis after transfecting Pinch-1 siRNA.

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Acknowledgments

The study was funded by grants from Military Natural Science foundation of China (06-MB238), University Natural Science Foundation of Third Military Medical University (2007XG33), Natural Science fund of Chongqing (2009C196) and National Natural Science Foundation of China (81000195).

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None.

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Authors and Affiliations

  1. Department of Hematology, Xinqiao Hospital, Third Military Medical University, Chongqing, 400037, China

    Dongfeng Zeng, Wei Xu, Zhihong Li, Jieping Li, Xi Zhang, Xinghua Chen & Peiyan Kong

  2. Department of Internal Medicine, No. 324 Hospital of PLA, Chongqing, 400020, China

    Lei Hao

  3. Department of Hematology, No. 52 Hospital of PLA, KangDing, SiChuan, China

    Weiyan Li

Authors
  1. Dongfeng Zeng
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  2. Lei Hao
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  3. Wei Xu
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  4. Zhihong Li
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  5. Weiyan Li
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  6. Jieping Li
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  7. Xi Zhang
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  8. Xinghua Chen
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  9. Peiyan Kong
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Corresponding author

Correspondence to Peiyan Kong.

Additional information

Dongfeng Zeng and Lei Hao contributed to this work equally.

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Zeng, D., Hao, L., Xu, W. et al. Pinch-1 was up-regulated in leukemia BMSC and its possible effect. Clin Exp Med 13, 21–27 (2013). https://doi.org/10.1007/s10238-012-0176-7

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  • DOI: https://doi.org/10.1007/s10238-012-0176-7

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