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.