Cancer microdissection

Ornstein DK et al. Proteomic analysis of laser capture microdissected human prostate cancer and in vitro prostate cell lines. Electrophoresis 2000 21 2235-2242. 

Specht et al.60 Microdissected cancer tissues RNA lysis buffer containing SDS and proteinase K Real time, for seven cancerrelevant genes Reproducible quantitation of specific mRNAs can be achieved... 

In 2004, Melle et al.12 proposed a technical triad of microdissection, pro-teomic techniques, and IHC that provides an integrated approach for characterization of cancer biomarkers. In recent years, a growing body of literature pertaining to this combined approach has accumulated. Table 22.1 shows a few selected examples under this topic. 

The term reverse phase protein microarray was first coined in 2001 by Paweletz et al. (5) in the landmark paper which introduced the platform and demonstrated its utility for analyzing proteins derived from laser capture microdissected (LCM) prostate cancer. RPMAs have also been referred to by other names in the literature including lysate arrays, reverse phase lysate arrays, protein microarrays, and reverse phase protein arrays (6). A very nice historical perspective on the development of RPJMA technology is presented in a review by Mueller et al. (6). A detailed description of the methods for performing RPMA is beyond the scope of this chapter and would be redundant because there are... 

Paweletz CP, Liotta LA, Petricorn EE (2001) New technologies for biomarker analysis of prostate cancer progression laser capture microdissection and tissue proteomics. Urology 57 160-163... 

VanMeter AJ, Rodriguez AS, Bowman ED et al (2008) Laser capture microdissection and protein microarray analysis of human non-smaU cell lung cancer differential epidermal growth factor receptor (EGPR) phosphorylation events associated with mutated EGFR compared with wild type. Mol Cell Proteomics 7 1902-1924... 

Kondo T, Seike M, Mori Y, Fujii K, Yamada T, Hirohashi S. (2003) Application of sensitive fluorescent dyes in linkage of laser microdissection and two-dimensional gel electrophoresis as a cancer proteomic study tool. Proteomics 3, 1758-66. 

While laser capture microdissection and other related techniques may allow for the generation of relatively homogenous cell populations, most such purification steps will largely eliminate any potential cancer stem cells, because these techniques necessitate the identification of differentiated cell types for isolation. As such, cancer stem cells would frequently be eliminated from analysis, and any cancer stem cells that may be accidentally included in the analysis would be in such low occurrence that their signature would most certainly be missed. 

Fend F, Raffeld M. Laser capture microdissection in pathology. J Clin Pathol 2000 53 666-672. Alevizos I, Mahadevappa M, Zhang X et al. Oral cancer in vivo gene expression profiling assisted by laser capture microdissection and microarray analysis. Oncogene 2001 20 6196-6204. 

Wu MS, Lin YS, Chang YT et al. Gene expression profiling of gastric cancer by microarray combined with laser capture microdissection. World J Gastroenterol 2005 11 7405-7412. 

Lieberfarb ME, Lin M, Lechpammer M et al. Genome-wide loss of heterozygosity analysis from laser capture microdissected prostate eaneer using single nucleotide polymorphic allele (SNP) arrays and a novel bioinformaties platform dChipSNP. Cancer Res 2003 63 4781 785. 

Moskaluk, C.A. (2001). Microdissection of histologic sections. Manual and laser capture microdissection techniques. In Powel, S.M., ed., Colorectal Cancer. Humana Press, Totowa, NJ, 1-13. 

Lawrie LC, Curran S. Laser capture microdissection and colorectal cancer proteomics. Methods Mol Biol 2005 293 245-253. 

Schuetz CS, Bonin M, Clare SE, Nieselt K, Sotlar K, Walter M, et al. Progression-specific genes identified by expression profiling of matched ductal carcinomas in situ and invasive breast tumors, combining laser capture microdissection and oligonucleotide microarray analysis. Cancer Res 2006 66(10) 5278—5286. 

Since tumor tissue is the primary manifestation of cancer, it contains the most immediate information and is the preferred material to investigate molecular targets. Experimental approaches will benefit from the enhanced protein concentration compared to body fluids. However, one major drawback of analyzing tumor biopsies is their limited availability and their heterogenicity which often requires microdissection. Alternatively, body fluids are the preferred clinical sources because they are most readily available in a noninvasive and continuous manner. 

Ray, M., Guan, X., Slovak, M Trent, J., and Meltzer, P. (1994) Rapid detection, cloning and molecular cytogenetic characterization of sequences from an MRP-encoding amplicon by chromosome microdissection. Br. J. Cancer 70, 85-90. 

Sugiyama, Y., Farrow, B., Murillo, C., Li, J., Watanabe, H., Sugiyama, K. and Evers, B.M. (2005) Analysis of differential gene expression patterns in colon cancer and cancer stroma using microdissected tissues. Gastroenterology, 128 (2), 480—486. 

Paweletz, C.P Liotta, L.A. Petricoin, E.F. New Technologies for Biomarker Analysis of Prostate Cancer Progression Laser Capture Microdissection and Tissue Proteomics, Urology 57,160-163 (2001). 

Key Words Cancer DNA laser capture microdissection molecular profiling proteomics protein RNA tissue heterogeneity. 

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