title: Analysis of microdissected cells by two-dimensional LC-MS approaches.
abstract: Laser capture microdissection (LCM) is a powerful tool that enables the isolation of specific cell types from tissue sections, overcoming the problem of tissue heterogeneity and contamination. We combined the LCM with isotope-coded affinity tag (ICAT) technology and two-dimensional liquid chromatography to investigate the qualitative and quantitative proteomes of hepatocellular carcinoma (HCC). The effects of three different histochemical stains on tissue sections have been compared, and toluidine blue stain was proved as the most suitable stain for LCM followed by proteomic analysis. The solubilized proteins from microdissected HCC and non-HCC hepatocytes were qualitatively and quantitatively analyzed with two-dimensional liquid chromatography tandem mass spectrometry (2D-LC-MS/MS) alone or coupled with cleavable isotope-coded affinity tag (cICAT) labeling technology. A total of 644 proteins were qualitatively identified and 261 proteins were unambiguously quantified. These results showed that the clinical proteomic method using LCM coupled with ICAT and 2D-LC-MS/MS can carry out not only large-scale but also accurate qualitative and quantitative analysis.
authors.affiliation: Research Center for Proteome Analysis, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China.
authors.forename: Chen
authors.initials: C
authors.lastname: Li
authors.lastname: Yi-Hong
authors.forename: Ye-Xiong
authors.initials: YX
authors.lastname: Tan
authors.forename: Jian-Hua
authors.initials: JH
authors.lastname: Ai
authors.forename: Hu
authors.initials: H
authors.lastname: Zhou
authors.forename: Su-Jun
authors.initials: SJ
authors.lastname: Li
authors.forename: Lei
authors.initials: L
authors.lastname: Zhang
authors.forename: Qi-Chang
authors.initials: QC
authors.lastname: Xia
authors.forename: Jia-Rui
authors.initials: JR
authors.lastname: Wu
authors.forename: Hong-Yang
authors.initials: HY
authors.lastname: Wang
authors.forename: Rong
authors.initials: R
authors.lastname: Zeng
mesh_terms: D006528:Carcinoma, Hepatocellular / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D019295:Computational Biology; D006801:Humans; D007553:Isotope Labeling / Q000379:methods; D007834:Lasers; D008113:Liver Neoplasms / Q000737:chemistry; D042282:Microdissection / Q000379:methods*; D009363:Neoplasm Proteins / Q000302:isolation & purification; D020543:Proteome / Q000302:isolation & purification; D040901:Proteomics / Q000379:methods* / Q000706:statistics & numerical data; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D009363:Neoplasm Proteins; D020543:Proteome