Figure 2.

Establishment of a highly lymphatic metastatic subline BxPC-3-LN including 5 steps: (1) subcutaneous footpad injection of nude mice with BxPC-3 cells; (2) doing autopsy to remove the possible metastatic lymph nodes which were confirmed by pathological confirmation; (3) isolation of metastatic cancer cells from identified lymph nodes; (4) selection and subculture of cells; (5) reinjection in nude mice for additional rounds and finally obtaining desired cell line BxPC-3-LN. And in the end, further analysis of two cell lines is urgently required such as comparison of gene and miRNAs expression profile via microarray.