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Silencing L-selectin expression by siRNA attenuated metastasis of murine lymphoid neoplasm cell P388D1 to peripheral lymph nodes

Leukemia volume 21, pages 180–183 (2007)Cite this article

About 50% of tumor metastasis is via the lymphatic system, but the underlying mechanisms have not been characterized. L-selectin (CD62L) is a lymphocyte homing receptor that is important for the migration of lymphocytes into peripheral lymph nodes.1 It is an intriguing question whether lymphoma cells use the same pathways of dissemination into peripheral lymph nodes.2, 3 In this work, we sought to ascertain whether the expression of L-selectin on macrophage-like lymphoid neoplasm P388D1 cells would facilitate their localization to peripheral lymph nodes.

Next, we investigated whether siRNA-induced L-selectin expression silencing affect P388D1 cells adhesion on to lymphatic endothelium in vivo. For this purpose, P388D1 cells were labeled with 5 μ M CFSE, a green fluorescence dye which can be transported across plasma membrane to react covalently with free amino group of intracellular macromolecules.5 In brief, two groups of DAB mice (n=6) received footpad injections of CFSE-tagged P388D1 cells transfected with L-selectin siRNA or control siRNA, 1 × 107 cells for each recipient. After 1−48 h, the mice were killed. Peripheral lymph nodes were removed and CFSE-tagged P388D1 cells in cell suspensions were evaluated by flow cytometry. A significant reduction of CFSE-tagged P388D1 cells population in popliteal nodes other than inguinal and axillary nodes was observed after 12 h in L-selectin siRNA-treated groups (Figure 2b). Similarly, two groups received footpad injections of CFSE-tagged P388D1 cells preincubated with 500 ng of MEL-14 or 500 ng of rat IgG, 1 × 107 cells for each recipient. In the MEL-14-treated group, CFSE-tagged P388D1 cells adhesion to popliteal and inguinal nodes but not axillary nodes was significantly lower than that in the control group at 12 h (Figure 2b). These data indicate that L-selectin on P388D1 cells plays an important role in adhesion to peripheral lymph nodes not only in vitro, but also in vivo.

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Acknowledgements

We thank Drs Jianxin Gu (Fudan University, Shanghai, China) and Haiying Chu, Li Jia and Huimin Zhou (Dalian Medical University, Dalian, China) for discussions, Yuejian Liu for technical assistance of flow cytometry analysis. This work was supported by grants from National Natural Science Foundation of China (30470400) and the Science and Technology Department of Liaoning Province, China.

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  1. Department of Biochemistry and Molecular Biology, Institute of Glycobiology, Dalian Medical University, Dalian, China

    Y Zuo, W Wei, C Liu, L Zhao & J Zhang

  2. Department of Clinical Biochemistry, Dalian Medical University, Dalian, China

    Y Zuo & W Wei

  3. Department of Pathology, the Second Affiliated Hospital of Dalian Medical University, Dalian, China

    L Wang

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  1. Y Zuo
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Zuo, Y., Wei, W., Liu, C. et al. Silencing L-selectin expression by siRNA attenuated metastasis of murine lymphoid neoplasm cell P388D1 to peripheral lymph nodes. Leukemia 21, 180–183 (2007). https://doi.org/10.1038/sj.leu.2404469

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