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Most solid cancers spread to new sites via the lymphatics before hematogenous dissemination. However, only a small fraction of an intravenously administered anti‐cancer drug enters the lymphatic system to reach metastatic lymph nodes ( LN ). Here, we show that the enhanced permeability and retention ( EPR ) effect is not induced during the early stages of  LN  metastasis. Luciferase‐expressing tumor cells were injected into the subiliac  LN  of the  MXH 10/Mo‐ lpr / lpr  mouse to induce metastasis to the proper axillary  LN  ( PALN ).  In vivo  biofluorescence imaging was used to confirm metastasis induction and to quantify the  EPR  effect, measured as  PALN  accumulation of intravenously injected indocyanine green ( ICG ) liposomes.  PALN  blood vessel volume changes were measured by contrast‐enhanced high‐frequency ultrasound imaging. The volume and density of blood vessels in the  PALN  increased until day 29 after inoculation, whereas the  LN  volume remained constant.  ICG  retention was first detected on day 29 post‐inoculation. While  CD 31‐positive cells increased up to day 29 post‐inoculation, α‐smooth muscle actin‐positive cells were detected on day 29 post‐inoculation for the first time. These results suggest that the  EPR  effect was not induced in the early stages of  LN  metastasis; therefore, systemic chemotherapy would likely not be beneficial during the early stages of  LN  metastasis. The development of an alternative drug delivery system, independent of the  EPR  effect, is required for the treatment of  LN  metastasis.

Evaluation of the enhanced permeability and retention effect in the early stages of lymph node metastasis