Proteomic analysis of primary tumor and lymph metastatic tissues as well as lung cancer cells

4 pairs of matched primary versus metastatic tumor tissues from formalin-fixed, paraffin-embedded (FFPE) tissue bank,and two frozen lung tumor tissues from patients with primary orlymph metastasis. Then performed tandem mass tag (TMT) labeling and quantitative mass spectrometry analysis. A549 Cells we used was overexpressed luceiferase and designated as L0,underwent 3rounds selection in vivo to form L2, L6 that were metastatic to the spinal. We cultured L0 cells in "light" medium, L2,L6 in "medium heavy" and "heavy" respectively. Harvesting of cells and combined in 1:1:1 ratio, peptides were seperated using high-PH RPLC and analyzed by LC-MS/MS.