Abstract
Gastrointestinal stromal tumors (GISTs) are the most common non-epithelial tumors in the digestive tract. Beyond surgery, therapeutic management depends on risk of recurrence. Risk evaluation of GIST takes into account location and size of the tumor, whether or not the tumor was intact or ruptured and mitotic index. The mitotic index lacks in intra- and interobserver reproducibility. In this study, we evaluated on 61 GISTs, the reproducibility of mitotic counting using classical hematoxylin-eosin-saffron (HES) staining, and its correlation with the mitotic count obtained through immunohistochemical staining for phospho-histone H3 (PHH3) and the proliferation index based upon Ki-67 immunostaining. Mitotic counts by HES and PHH3 staining and Ki-67 proliferation index were evaluated twice by three independent observers taking into account interpretation times per tumor for each technique. HES-based and PHH3-based mitotic counts and Ki-67 proliferation index correlated well and presented good intra- and interobserver reproducibility. PHH3 staining resulted in a slight but statistically significant difference of about two more mitotic figures per 5 mm2 than the HES-based count, which might have put some borderline tumors in a different risk category. Immunohistochemical staining for PHH3 and Ki-67 allowed more rapid interpretation than mitotic counts based upon HES staining, but only PHH3 staining allows counting of mitoses. Immunostaining using anti-PHH3 and anti-Ki-67 antibodies will eventually provide improved recurrence risk stratification of GIST and may become effective ancillary tools in deciding on optimal therapeutic management.
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References
Tryggvason G, Gislason HG, Magnusson MK, Jonasson JG (2005) Gastrointestinal stromal tumors in Iceland, 1990–2003: the Icelandic GIST study, a population-based incidence and pathologic risk stratification study. Int J Cancer 117:289–293
Miettinen M, Lasota J (2013) Gastrointestinal stromal tumors. Gastroenterol Clin N Am 42:399–415
Huizinga JD, Thuneberg L, Kluppel M, Malysz J, Mikkelsen HB, Bernstein A (1995) W/kit gene required for interstitial cells of Cajal and for intestinal pacemaker activity. Nature 373:347–349
Corless CL, Barnett CM, Heinrich MC (2011) Gastrointestinal stromal tumours: origin and molecular oncology. Nat Rev Cancer 11:865–878
Rubin BP, Singer S, Tsao C, Duensing A, Lux ML, Ruiz R, Hibbard MK, Chen CJ, Xiao S, Tuveson DA, Demetri GD, Fletcher CD, Fletcher JA (2001) KIT activation is a ubiquitous feature of gastrointestinal stromal tumors. Cancer Res 61:8118–8121
Corless CL, Schroeder A, Griffith D, Town A, McGreevey L, Harrell P, Shiraga S, Bainbridge T, Morich J, Heinrich MC (2005) PDGFRA mutations in gastrointestinal stromal tumors: frequency, spectrum and in vitro sensitivity to imatinib. J Clin Oncol 23:5357–5364
Debiec-Rychter M, Sciot R, Le CA, Schlemmer M, Hohenberger P, van Oosterom AT, Blay JY, Leyvraz S, Stul M, Casali PG, Zalcberg J, Verweij J, Van GM, Hagemeijer A, Judson I (2006) KIT mutations and dose selection for imatinib in patients with advanced gastrointestinal stromal tumours. Eur J Cancer 42:1093–1103
Novelli M, Rossi S, Rodriguez-Justo M, Taniere P, Seddon B, Toffolatti L, Sartor C, Hogendoorn PC, Sciot R, Van GM, Verweij J, Blay JY, Hohenberger P, Flanagan A, Dei Tos AP (2010) DOG1 and CD117 are the antibodies of choice in the diagnosis of gastrointestinal stromal tumours. Histopathology 57:259–270
Fletcher CD, Berman JJ, Corless C, Gorstein F, Lasota J, Longley BJ, Miettinen M, O'Leary TJ, Remotti H, Rubin BP, Shmookler B, Sobin LH, Weiss SW (2002) Diagnosis of gastrointestinal stromal tumors: a consensus approach. Int J Surg Pathol 10:81–89
Joensuu H (2008) Risk stratification of patients diagnosed with gastrointestinal stromal tumor. Hum Pathol 39:1411–1419
Miettinen M, Lasota J (2006) Gastrointestinal stromal tumors: pathology and prognosis at different sites. Semin Diagn Pathol 23:70–83
Casali PG, Fumagalli E, Gronchi A (2012) Adjuvant therapy of gastrointestinal stromal tumors (GIST). Curr Treat Options Oncol 13:277–284
Reichardt P, Blay JY, Boukovinas I, Brodowicz T, Broto JM, Casali PG, Decatris M, Eriksson M, Gelderblom H, Kosmidis P, Le CA, Pousa AL, Schlemmer M, Verweij J, Joensuu H (2012) Adjuvant therapy in primary GIST: state-of-the-art. Ann Oncol 23:2776–2781
Angi M, Damato B, Kalirai H, Dodson A, Taktak A, Coupland SE (2011) Immunohistochemical assessment of mitotic count in uveal melanoma. Acta Ophthalmol 89:e155–e160
Polley MY, Leung SC, McShane LM, Gao D, Hugh JC, Mastropasqua MG, Viale G, Zabaglo LA, Penault-Llorca F, Bartlett JM, Gown AM, Symmans WF, Piper T, Mehl E, Enos RA, Hayes DF, Dowsett M, Nielsen TO (2013) An international Ki67 reproducibility study. J Natl Cancer Inst 105:1897–1906
Hasegawa T, Yamamoto S, Nojima T, Hirose T, Nikaido T, Yamashiro K, Matsuno Y (2002) Validity and reproducibility of histologic diagnosis and grading for adult soft-tissue sarcomas. Hum Pathol 33:111–115
Yamaguchi U, Hasegawa T, Sakurai S, Sakuma Y, Takazawa Y, Hishima T, Mitsuhashi T, Sekine S, Chuman H, Shimoda T (2006) Interobserver variability in histologic recognition, interpretation of KIT immunostaining, and determining MIB-1 labeling indices in gastrointestinal stromal tumors and other spindle cell tumors of the gastrointestinal tract. Appl Immunohistochem Mol Morphol 14:46–51
Colman H, Giannini C, Huang L, Gonzalez J, Hess K, Bruner J, Fuller G, Langford L, Pelloski C, Aaron J, Burger P, Aldape K (2006) Assessment and prognostic significance of mitotic index using the mitosis marker phospho-histone H3 in low and intermediate-grade infiltrating astrocytomas. Am J Surg Pathol 30:657–664
Duregon E, Molinaro L, Volante M, Ventura L, Righi L, Bolla S, Terzolo M, Sapino A, Papotti MG (2014) Comparative diagnostic and prognostic performances of the hematoxylin-eosin and phospho-histone H3 mitotic count and Ki-67 index in adrenocortical carcinoma. Mod Pathol 27:1246–1254
Fukushima S, Terasaki M, Sakata K, Miyagi N, Kato S, Sugita Y, Shigemon M (2009) Sensitivity and usefulness of anti-phosphohistone-H3 antibody immunostaining for counting mitotic figures in meningioma cases. Brain Tumor Pathol 26:51–57
Hale CS, Qian M, Ma MW, Scanlon P, Berman RS, Shapiro RL, Pavlick AC, Shao Y, Polsky D, Osman I, Darvishian F (2013) Mitotic rate in melanoma: prognostic value of immunostaining and computer-assisted image analysis. Am J Surg Pathol 37:882–889
Kemmerling R, Weyland D, Kiesslich T, Illig R, Klieser E, Jager T, Dietze O, Neureiter D (2014) Robust linear regression model of Ki-67 for mitotic rate in gastrointestinal stromal tumors. Oncol Lett 7:745–749
Skaland I, Janssen EA, Gudlaugsson E, Klos J, Kjellevold KH, Soiland H, Baak JP (2007) Phosphohistone H3 expression has much stronger prognostic value than classical prognosticators in invasive lymph node-negative breast cancer patients less than 55 years of age. Mod Pathol 20:1307–1315
Tsuta K, Liu DC, Kalhor N, Wistuba II, Moran CA (2011) Using the mitosis-specific marker anti-phosphohistone H3 to assess mitosis in pulmonary neuroendocrine carcinomas. Am J Clin Pathol 136:252–259
Veras E, Malpica A, Deavers MT, Silva EG (2009) Mitosis-specific marker phospho-histone H3 in the assessment of mitotic index in uterine smooth muscle tumors: a pilot study. Int J Gynecol Pathol 28:316–321
Zhu S, Lin F, Chen ZE (2013) Mitosis-specific marker PPH3 immunostain is a more sensitive and efficient method to evaluate the mitotic activity in gastrointestinal stromal tumor (GIST). Lab Investig 93:190A
Hendzel MJ, Wei Y, Mancini MA, Van HA, Ranalli T, Brinkley BR, Bazett-Jones DP, Allis CD (1997) Mitosis-specific phosphorylation of histone H3 initiates primarily within pericentromeric heterochromatin during G2 and spreads in an ordered fashion coincident with mitotic chromosome condensation. Chromosoma 106:348–360
Hendzel MJ, Nishioka WK, Raymond Y, Allis CD, Bazett-Jones DP, Th'ng JP (1998) Chromatin condensation is not associated with apoptosis. J Biol Chem 273:24470–24478
Juan G, Traganos F, James WM, Ray JM, Roberge M, Sauve DM, Anderson H, Darzynkiewicz Z (1998) Histone H3 phosphorylation and expression of cyclins A and B1 measured in individual cells during their progression through G2 and mitosis. Cytometry 32:71–77
Casper DJ, Ross KI, Messina JL, Sondak VK, Bodden CN, McCardle TW, Glass LF (2010) Use of anti-phosphohistone H3 immunohistochemistry to determine mitotic rate in thin melanoma. Am J Dermatopathol 32:650–654
Idriss MH, Kazlouskaya V, Malhotra S, Andres C, Elston DM (2013) Phosphohistone-H3 and Ki-67 immunostaining in cutaneous pilar leiomyoma and leiomyosarcoma (atypical intradermal smooth muscle neoplasm). J Cutan Pathol 40:557–563
Jiang J, Jin MS, Suo J, Wang YP, He L, Cao XY (2012) Evaluation of malignancy using Ki-67, p53, EGFR and COX-2 expressions in gastrointestinal stromal tumors. World J Gastroenterol 18:2569–2575
Liang YM, Li XH, Li WM, Lu YY (2012) Prognostic significance of PTEN, Ki-67 and CD44s expression patterns in gastrointestinal stromal tumors. World J Gastroenterol 18:1664–1671
Molenaar WM, Plaat BE, Berends ER, te Meerman GJ (2000) Observer reliability in assessment of mitotic activity and MIB-1-determined proliferation rate in pediatric sarcomas. Ann Diagn Pathol 4:228–235
Acknowledgments
This study was supported by “Omnium group.” We would like to also acknowledge the Brest Hospital Biobank and Mrs Morgan Riou for their contribution in this study.
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The authors declare that they have no conflict of interest.
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Uguen, A., Conq, G., Doucet, L. et al. Immunostaining of phospho-histone H3 and Ki-67 improves reproducibility of recurrence risk assessment of gastrointestinal stromal tumors. Virchows Arch 467, 47–54 (2015). https://doi.org/10.1007/s00428-015-1763-2
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DOI: https://doi.org/10.1007/s00428-015-1763-2