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Drug response analysis of non-small cell lung cancer cells by phosphoproteome analysis
Mass spectrometry-based proteomics is an indispensable tool for molecular, cellular biology and for the emerging field of clinical medicine. Recently, we have developed a key technology on peptide separation for rapid and sensitive proteome analysis 1). We also developed a sensitive method to identify phospho tyrosine proteome using an immune affinity enrichment method 2).
Combining these technologies, we performed temporal characterization of non-small-cell lung cancer cell lines treated with erlotinib. We obtained phosphoproteome and phosphotyrosine-proteome profiles of two erlotinib-sensitive cells and four erlotinib-resistant cells treated by erlotinib for 0 h, 6 h and 24 h.
We quantified over 12000 phosphorylation sites including 600 phosphorylation sites on tyrosine residue across six cell lines. Especially, phosphotyrosine-proteome can detect bypass kinases which were known to be related to the drug resistance, such as FGFR, AXL, MET. Then, we extracted kinases and other enzymes which are up-regulated in resistant cells and selected 46 inhibitors for drug screening. 24 of 46 inhibitors inhibited cell growth of at least one resistant cell line. Phosphoproteomic approach is applicable for the detection of the markers for drug-efficacy and the target candidates for overcoming drug resistance.
References
1) J. Adachi et al., Anal. Chem., 88, 7899–7903 (2016).
2) Y. Abe et al., J. Proteome Res., 16, 1077–1086 (2017).