A PRELIMINARY STUDY ON VASCULAR ENDOTHELIAL GROWTH FACTOR C GENE MODIFIED LYMPH NODE TRANSPLANTATION IN PROMOTING PROLIFERATION OF LYMPHATIC ENDOTHELIAL CELLS_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

LU Nanhang , ZHANG Yong , FENG Zihao , QI Fazhi.

Department of Plastic &;
Reconstructive Surgery, Zhongshan Hospital, Fudan University, Shanghai, 200032, P.R.China. Corresponding author: QI Fazhi, E-mail: qi.fazhi@zs-hospital.sh.cn;

Corresponding author：

Keywords：

Lymph node transplantation; Vascular endothelial growth factor; C Lymphatic endothelial cells; Rat

DOI：

10.7507/1002-1892.20130136

Video：

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Objective To investigate the effects of vascular endothelial growth factor C (VEGF-C) gene modified lymph nodes on promoting proliferation of lymphatic endothelial cells in the surrounding tissues. Methods Thirty-six Sprague Dawley rats, weighing 200.1-271.5 g, were randomly divided into 2 groups (n=18). After the in situ axillary lymph nodes transplantation models were established in both groups, 1.5 × 108 PFU Ad-VEGF-C-Flag and Ad-Flag were injected into the transplanted lymph nodes in experimental group and control group, respectively. At 3 days after injection, the axillary lymph nodes were harvested to observe the expression of Flag; at 1, 2, and 4 weeks after injection, the axillary lymph nodes and the surrounding tissues were harvested to observe the expression of Prxo-1 protein and to calculate the fluorescence density; at 2 and 4 weeks after injection, the absorbance (A) value of treated blood at 620 nm was calculated to observe lymphatic back-flow function improvement; the rats without treatment served as normal control group, and the rats with in situ axillary lymph nodes transplantation model served as blank control group. Results At 3 days after injection, the expression of Flag could be detected in experimental group and control group. The fluorescence density of Prox-1 protein in experimental group increased at 1, 2, and 4 weeks, and it was significantly higher than that in control group (P lt; 0.05). The A values of normal control group and blank control group were 0.539 ± 0.020 and 0.151 ± 0.007, respectively. The A values of experimental group and control group were 0.170 ± 0.011 and 0.168 ± 0.010 at 2 weeks, and 0.212 ± 0.016 and 0.197 ± 0.006 at 4 weeks, which were significantly lower than those of normal control group (P lt; 0.05), but no significant difference was found when compared with blank control group, and between the experimental group and control group (P gt; 0.05). Conclusion The VEGF-C gene modified lymph nodes can stimulate the proliferation of lymphatic endothelial cells in the surrounding tissues. However, it has no improved effect on lymphatic back-flow function in the affected limb.

Citation： LU Nanhang,ZHANG Yong,FENG Zihao,QI Fazhi.. A PRELIMINARY STUDY ON VASCULAR ENDOTHELIAL GROWTH FACTOR C GENE MODIFIED LYMPH NODE TRANSPLANTATION IN PROMOTING PROLIFERATION OF LYMPHATIC ENDOTHELIAL CELLS. Chinese Journal of Reparative and Reconstructive Surgery, 2013, 27(5): 619-623. doi: 10.7507/1002-1892.20130136 Copy

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1. Oremus M, Dayes I, Walker K, et al. Systematic review: conservative treatments for secondary lymphedema. BMC Cancer, 2012, 12: 6.
2. Lin CH, Ali R, Chen SC, et al. Vascularized groin lymph node transfer using the wrist as a recipient site for management of postmastectomy upper extremity lymphedema. Plast Reconstr Surg, 2009, 123(4): 1265-1275.
3. Becker C, Assouad J, Riquet M, et al. Postmastectomy lymphedema: long-term results following microsurgical lymph node transplantation. Ann Surg, 2006, 243(3): 313-315.
4. Saaristo AM, Niemi TS, Viitanen TP, et al. Microvascular breast reconstruction and lymph node transfer for postmastectomy lymphedema patents. Ann Surg, 2012, 255(3): 468-473.
5. Karkkainen MJ, Haiko P, Sainio K, et al. Vascular endothelial growth factor C is required for sprouting of the first lymphatic vessels from embryonic veins. Nat Immunol, 2004, 5(1): 74-80.
6. Küchler AM, Gjini E, Peterson-Maduro J, et al. Development of the zebrafish lymphatic system requires VEGF-C signaling. Curr Biol, 2006, 16(12): 1244-1248.
7. Yaniv K, Isogai S, Castranova D, et al. Live imaging of lymphatic development in the zebrafish. Nat Med, 2006, 12(6): 711-716.
8. Shin WS, Rockson SG. Animal models for the molecular and mechanistic study of lymphatic biology and disease. Ann N Y Acad Sci, 2008, 1131: 50-74.
9. Kanter MA, Slavin SA, Kaplan W. An experimental model for chronic lymphedema. Plast Reconstr Surg, 1990, 85(4): 573-580.
10. Shin WS, Szuba A, Rockson SG. Animal models for the study of lymphatic insufficiency. Lymphat Res Biol, 2003, 1(2): 159-169.
11. Tammela T, Saaristo A, Holopainen T, et al. Therapeutic differentiation and maturation of lymphatic vessels after lymph node dissection and transplantation. Nat Med, 2007, 13(12): 1458-1466.
12. Alitalo K, Tammela T, Petrocva TV. Lymphangiogenesis in development and human disease. Nature, 2005, 438(7070): 946-953.
13. Karkkainen MJ, Haiko P, Sainio K, et al. Vascular endothelial growth factor C is required for sprouting of the first lymphatic vessels from embryonic veins. Nat Immunol, 2004, 5(1): 74-80.
14. Foster DN, Min B, Foster LK, et al. Positive and negative cis-acting regulatory elements mediate expression of the mouse vascular smooth muscle alpha-actin gene. J Biol Chem, 1992, 267 (17): 11995-12003.
15. Mumprecht V, Roudnicky F, Detmar M. Inflammation-induced lymph node lymphangiogenesis is reversible. Am J Pathol, 2012, 180(3): 874-879.
16. Alam A, Herault JP, Barron P, et al. Heterodimerization with vascular endothelial growth factor receptor-2 (VEGFR-2) is necessary for VEGFR-3 activity. Biochem Biophys Res Commun, 2004, 324(2): 909-915.
17. Tammela T, Alitalo K. Lymphangiogenesis: Molecular mechanisms and future promise. Cell, 2010, 140(4): 460-476.
18. Song SH, Kim KL, Lee KA, et al. Tie1 regulates the Tie2 agonistic role of angiopoietin-2 in human lymphatic endothelial cells. Biochem Biophys Res Commun, 2012, 419(2): 281-286.
19. Nguyen VP, Chen SH, Trinh J, et al. Differential response of lymphatic, venous and arterial endothelial cells to angiopoietin-1 and angiopoietin-2. BMC Cell Biol, 2007, 8: 10.
20. Shioda H. Immunohistochemical demonstration of Angiopoietin-2 in lymphatic vascular development. Histochem Cell Biol, 2009, 131(2): 231-238.
21. Wu X, Liu NF. FOXC2 transcription factor: a novel regulator of lymphangiogenesis. Lymphology, 2011, 4(1): 35-41.
22. Hayashi H, Kume T. Foxc2 transcription factor as a regulator of angiogenesis via induction of integrin beta3 expression. Cell Adh Migr, 2009, 3(1): 24-26.
23. Sabine A, Agalarov Y, Maby-El Hajjami H, et al. Mechanotransduction, PROX1, and FOXC2 cooperate to control connexin37 and calcineurin during lymphatic-valve formation. Dev Cell, 2012, 22(2): 430-445.

Chinese Journal of Reparative and Reconstructive Surgery

A PRELIMINARY STUDY ON VASCULAR ENDOTHELIAL GROWTH FACTOR C GENE MODIFIED LYMPH NODE TRANSPLANTATION IN PROMOTING PROLIFERATION OF LYMPHATIC ENDOTHELIAL CELLS

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