Abstract

Background: Most bladder cancer patients experience lymphatic metastasis in the course of disease progression,
yet the relationship between lymphangiogenesis and lymphatic metastasis is not well known. The aim of this study
is to elucidate underlying mechanisms of how expanded lymphatic vessels and tumor microenvironment interacts
each other and to find effective therapeutic options to inhibit lymphatic metastasis.
Results: The orthotopic urinary bladder cancer (OUBC) model was generated by intravesical injection of MBT-2 cell
lines. We investigated the angiogenesis, lymphangiogenesis, and CD11b+/CD68+ tumor-associated macrophages
(TAM) by using immunofluorescence staining. OUBC displayed a profound lymphangiogenesis and massive
infiltration of TAM in primary tumor and lymphatic metastasis in lymph nodes. TAM flocked near lymphatic vessels
and express higher levels of VEGF-C/D than CD11b- cells. Because VEGFR-3 was highly expressed in lymphatic
vascular endothelial cells, TAM could assist lymphangiogenesis by paracrine manner in bladder tumor. VEGFR-3
expressing adenovirus was administered to block VEGF-C/D signaling pathway and clodronate liposome was used
to deplete TAM. The blockade of VEGF-C/D with soluble VEGF receptor-3 markedly inhibited lymphangiogenesis
and lymphatic metastasis in OUBC. In addition, the depletion of TAM with clodronate liposome exerted similar
effects on OUBC.
Conclusion: VEGF-C/D are the main factors of lymphangiogenesis and lymphatic metastasis in bladder cancer.
Moreover, TAM plays an important role in these processes by producing VEGF-C/D. The inhibition of
lymphangiogenesis could provide another therapeutic target to inhibit lymphatic metastasis and recurrence in
patients with invasive bladder cancer.