Summary
Tumors are heterogeneous structures that contain different cell populations. Laser capture microdissection (LCM) can be used to obtain pure cancer cells from fresh-frozen cancer tissue and the surrounded environment, thus providing an accurate snapshot of the tumor and its microenvironment in vivo. We describe a new approach to isolate pure cancer cell population and evaluate protein expression. The process includes immunocytochemistry, laser microdissection, and western blot analysis. Using this technique, we can detect proteins such as X-linked inhibitor of apoptosis protein (XIAP) and Fas ligand (FasL) with as little as 1000 cells.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Jones MB, Krutzsch H, Shu H, Zhao Y, Liotta LA, Kohn EC, Petricoin EF 3rd. Proteomic analysis and identification of new biomarkers and therapeutic targets for invasive ovarian cancer. Proteomics 2(1):76–84, 2002.
Agiostratidou G, Sgouros I, Galani E, Voulgari A, Chondrogianni N, Samantas E, Dimopoulos MA, Skarlos D, Gonos ES. Correlation of in vitro cytotoxicity and clinical response to chemotherapy in ovarian and breast cancer patients. Anticancer Research 21(1A):455–9, 2001.
Emmert-Buck MR, Bonner RF, Smith PD, Chuaqui RF, Zhuang Z, Goldstein SR, Weiss RA, Liotta LA. Laser capture microdissection. Science 274(5289):998–1001, 1996.
Wang VW, Bell DA, Berkowitz RS, Mok SC. Whole genome amplification and high-throughput allelotyping identified five distinct deletion regions on chromosomes 5 and 6 in microdissected early-stage ovarian tumors. Cancer Research 61(10):4169–74, 2001.
Jemal A, Murray T, Samuels A, Ghafoor A, Ward E, Thun MJ. Cancer statistics, 2003. CA Cancer J Clin 2003;53:5–26.
Hoskins WJ, Perez CA, Young RC, Barakat RR, Markman M, Randall ME. Principles and Practice of Gynecologic Oncology. 4th ed. 2005.
Sapi E, Alvero AB, Chen W, O’Malley D, Hao XY, Dwipoyono B, Garg M, Kamsteeg M, Rutherford T, Mor G. Resistance of ovarian carcinoma cells to docetaxel is XIAP dependent and reversible by phenoxodiol. Oncology Research 14(11–12):567–78, 2004.
Abrahams VM, Straszewski SL, Kamsteeg M, Hanczaruk B, Schwartz PE, Rutherford TJ, Mor G. Epithelial ovarian cancer cells secrete functional Fas ligand. Cancer Research 63(17):5573–81, 2003.
Abrahams VM, Kamsteeg M, Mor G. The Fas/Fas ligand system and cancer: immune privilege and apoptosis. Molecular Biotechnology 25(1):19–30, 2003.
Hardiman G, Jenkins NA, Copeland NG, Gilbert DJ, Garcia DK, Naylor SL, Kastelein RA, Bazan JF. Genetic structure and chromosomal mapping of MyD88. Genomics 45(2):332–9, 1997.
Akazawa T, Masuda H, Saeki Y, Matsumoto M, Takeda K, Tsujimura K, Kuzushima K, Takahashi T, Azuma I, Akira S, Toyoshima K, Seya T. Adjuvant-mediated tumor regression and tumor-specific cytotoxic response are impaired in MyD88-deficient mice. Cancer Research 64(2):757–64, 2004.
Zeisel MB, Druet VA, Sibilia J, Klein JP, Quesniaux V, Wachsmann D. Cross talk between MyD88 and focal adhesion kinase pathways. Journal of Immunology 174(11):7393–7, 2005.
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2008 Humana Press Inc.
About this protocol
Cite this protocol
Silasi, DA. et al. (2008). Detection of Cancer-Related Proteins in Fresh-Frozen Ovarian Cancer Samples Using Laser Capture Microdissection. In: Mor, G., Alvero, A.B. (eds) Apoptosis and Cancer. Methods in Molecular Biology™, vol 414. Humana Press. https://doi.org/10.1007/978-1-59745-339-4_4
Download citation
DOI: https://doi.org/10.1007/978-1-59745-339-4_4
Publisher Name: Humana Press
Print ISBN: 978-1-58829-457-9
Online ISBN: 978-1-59745-339-4
eBook Packages: Springer Protocols