Chromogenic in situ hybridization (CISH): a novel alternative in screening archival breast cancer tissue samples for HER-2/neu status

Breast Cancer Research

Raymundo W Lo¹  Manuelito A Madrid¹ 

[1] Institute of Pathology, St Luke's Medical Center, Quezon City, Philippines

关键词: immunohistochemistry;    Her-2/neu;    fluorescence in situ hybridization;    chromogenic in situ hybridization;    breast cancer;   

Others  :  1118722
DOI  :  10.1186/bcr915

 received in 2004-04-23, accepted in 2004-06-29,  发布年份 2004

【 摘 要 】

Background

Chromogenic in situ hybridization (CISH) is emerging as a practical, cost-effective, and valid alternative to fluorescent in situ hybridization in testing for gene alteration, especially in centers primarily working with immunohistochemistry (IHC).

Methods

We assessed Her-2/neu alteration using CISH on formalin-fixed paraffin-embedded primary invasive ductal carcinoma tumors in which IHC (CB11 antibody) had previously been performed, and we compared the results with IHC. The 160 selected cases were equally stratified randomly into the four IHC categories (scores of 0, 1+, 2+, and 3+). We also compared age at diagnosis and tumor histologic grade with IHC and CISH Her-2/neu.

Results

We were able to perform and evaluate CISH successfully on all cases. The agreement between 3+ IHC and CISH-amplified cases as well as between all IHC and CISH Her-2/neu negative cases was 100%, and the concordance on all positive cases was 72.50%, with an overall agreement of 86.25%. All the discordant cases had 2+ IHC scores. Although we noted Her-2/neu positivity more in premenopausal women, the age at diagnosis was not significantly associated with IHC or CISH results. Similarly, although the small group of well-differentiated tumors was apparently Her-2/neu negative in both tests, no significant association was noted between any tumor histologic grade and either IHC or CISH results.

Conclusions

CISH is easily integrated into routine testing in our laboratory. It is a necessary adjunct in determining the subset of non-amplified IHC-positive invasive tumors that will not benefit from trastuzumab therapy. Those cases with 2+ IHC results will be triaged and subjected to CISH. Her-2/neu testing should be done on all breast cancer cases regardless of age at presentation and tumor histologic grade.

【 授权许可】

   
2004 Madrid and Lo; licensee BioMed Central Ltd. This is an Open Access article: verbatim copying and redistribution of this article are permitted in all media for any purpose, provided this notice is preserved along with the article's original URL.

【 参考文献 】

• [1]Popescu NC, King CR, Kraus MH: Localization of the human erbB-2 gene on normal and rearranged chromosomes 17 to bands q12-21.32. Genomics 1989, 4:362-366.
• [2]Akiyama T, Sudo C, Ogawara H, Toyoshima K, Yamamoto T: The product of c-erbB-2 gene. A 185-kilodalton glycoprotein with tyrosine kinase activity. Science 1986, 232:1644-1646.
• [3]Ross JS, Fletcher JA: HER-2/neu (C-erb-B2) gene and protein in breast cancer. Am J Clin Pathol 1999, 112:53-67.
• [4]Slamon DJ, Clark GM, Wong SG, Levin WJ, Ullrich A, McGuire WL: Human breast cancer: correlation of relapse and survival with amplification of the Her-2/neu oncogene. Science 1987, 235:177-182.
• [5]Sjogren S, Inganas M, Lingren A, Holmberg L, Bergh J: Prognostic and predictive value of c-erbB-2 overexpression in primary breast cancer, alone and in combination with other prognostic markers. J Clin Oncol 1998, 16:462-469.
• [6]Borg A, Baldetorp B, Ferno M, Kilander D, Olsson H, Ryden S, Sigurdsson H: ERBB2 amplification is associated with tamoxifen resistance in steroid-receptor positive breast cancer. Cancer Lett 1994, 81:137-144.
• [7]Carlomagno C, Perrone F, Gallo C, De Laurentiis M, Lauria R, Morabito A, Pettinato G, Panico L, D'Antonio A, Bianco AR, DePlacido S: c-erbB-2 overexpression decreases the benefit of adjuvant tamoxifen in early stage breast cancer without axillary lymph node metastases. J Clin Oncol 1996, 14:2702-2708.
• [8]Tetu B, Brisson J, Plante V, Bernard P: p53 and c-erbB-2 as markers of resistance to adjuvant chemotherapy in breast cancer. Mod Pathol 1998, 11:823-830.
• [9]Cobleigh MA, Vogel CL, Tripathy D, Robert NJ, Scholl S, Fehrenbacher L, Wolter JM, Paton V, Shak S, Lieberman G, Slamon DJ: Multinational study of the efficacy and safety of humanized anti-HER2-overexpressing metastatic breast cancer that has progressed after chemotherapy for metastatic disease. J Clin Oncol 1999, 17:2639-2348.
• [10]Jacobs TW, Gown AM, Yaziji H, Barnes MJ, Schnitt SJ: Specificity of HercepTest in determining Her-2/neu status of breast cancers using the United States Food and Drug Administration-approved scoring system. J Clin Oncol 1999, 17:1983-1987.
• [11]Tubbs RR, Pettay JD, Roche PC, Stoler MH, Jenkins RB, Grogan TM: Discrepancies in clinical laboratory testing of eligibility for trastuzumab therapy; apparent immunohistochemical false-positive do not get the message. J Clin Oncol 2001, 19:2714-2721.
• [12]Park K, Kim J, Lim S, Han S, Lee JY: Comparing fluorescence in situ hybridization and chromogenic in situ hybridization methods to determine the Her2/neu status in primary breast carcinoma using tissue microarray. Mod Pathol 2003, 26:937-943.
• [13]Elston CW, Ellis IO: Pathological prognostic factors in breast cancer: I. The value of histologic grade in breast cancer – experience from a large study with long-term follow-up. Histopathology 1991, 19:403-310.
• [14]Corbett IP, Henry JA, Angus B, Watchorn CJ, Wilkinson L, Hennessy C, Gullick WJ, Tuzi NL, May FE, Westley BR, et al.: NCL-CB11, a new monoclonal antibody recognizing the internal domain of the c-erbB-2 oncogene protein effective for use on formalin-fixed, paraffin-embedded tissue. J Pathol 1990, 161:15-25.
• [15]Press MF, Hung G, Godolphin G, Slamon DJ: Sensitivity of Her-2/neu antibodies in archival tissue samples: potential source of error in immunohistochemical studies of oncogene expression. Cancer Res 1994, 54:2771-2777.
• [16]Thomson TA, Hayes MM, Spinelli JJ, Hilland E, Sawrenko C, Phillips D, Dupuis B, Parker RL: Her-2/neu in breast cancer: interobserver variability and performance of immunohistochemistry with 4 antibodies compared with fluorescent in situ hybridization. Mod Pathol 2001, 14:1079-1086.
• [17]Couturier J, Vincent-Salomon A, Nicolas A, Beuzeboc P, Mouret E, Zafrani B, Sastre-Garau X: Strong correlation between results of fluorescent in situ hybridization and immunohistochemistry for the assessment of the ERBB2 (Her-2/neu) gene status in breast carcinoma. Mod Pathol 2000, 13:1238-1243.
• [18]Jimenez RE, Wallis T, Tabasczka P, Visscher DW: Determination of Her-2/neu status in breast carcinoma: comparative analysis of immunohistochemistry and fluorescent in situ hybridization. Mod Pathol 2000, 13:37-45.
• [19]Pauletti G, Godolphin W, Press MF: Detection and quantification of Her2/neu gene amplification in human breast cancer archival material using fluorescence in situ hybridization. Oncogene 1996, 13:63-72.
• [20]Kakar S, Puangsuvan N, Stevens JM, Serenas R, Mangan G, Sahai S, Mihalov ML: Her-2/neu assessment in breast cancer by immunohistochemistry and fluorescence in situ hybridization: comparison of results and correlation with survival. Mol Diagn 2000, 5:199-207.
• [21]Tanner M, Gancberg D, Di Leo A, Larsimont D, Rouas G, Piccart MJ, Isola J: Chromogenic in situ hybridization: a practical alternative for fluorescence in situ hybridization to detect HER-2/neu oncogene amplification in archival breast cancer samples. Am J Pathol 2000, 157:1467-1472.
• [22]Kumamoto H, Sasano H, Taniguchi T, Suzuki T, Moriya T, Ichinohasama R: Chromogenic in situ hybridization analysis of HER-2/neu status in breast carcinoma: application in screening of patients for trastuzumab (Herceptin) therapy. Pathol Int 2001, 51:579-584.
• [23]Dandachi N, Dietze O, Hauser-Kronberger C: Chromogenic in situ hybridization: a novel approach to a practical and sensitive method for the detection of HER2 oncogene in archival human breast carcinoma. Lab Invest 2002, 82:1007-1014.
• [24]Zhao J, Wu R, Au A, Marquez A, Yu Y, Shi Z: Determination of Her2 gene amplification by chromogenic in situ hybridization (CISH) in archival breast carcinoma. Mod Pathol 2002, 15:657-665.
• [25]Gupta D, Middleton LP, Whitaker MJ, Abrams J: Comparison of fluorescence and chromogenic in situ hybridization for detection of Her-2/neu oncogene in breast cancer. Am J Clin Pathol 2003, 119:381-387.
• [26]Arnould L, Denoux Y, MacGrogan G, Penault-Llorca F, Fiche M, Treilleux I, Mathieu MC, Vincent-Salomon A, Vilain MO, Couturier J: Agreement between chromogenic in situ hybridization (CISH) and FISH in the determination of Her2 status in breast cancer. Br J Cancer 2003, 88:1587-1591.
• [27]Park K, Kim J, Lim S, Han S, Lee JY: Comparing fluorescence in situ hybridization and chromogenic in situ hybridization methods to determine the Her2/neu status in primary breast carcinoma using tissue microarray. Mod Pathol 2003, 26:937-943.
• [28]Vera-Roman JM, Rubio-Martinez LA: Comparative assays for the HER-2/neu oncogene status in breast cancer. Arch Pathol Lab Med 2004, 128:627-633.
• [29]Ross JS, Fletcher JA, Bloom KJ, Linette GP, Stec J, Clark E, Ayers M, Symmans WF, Pusztai L, Hortobagyi GN: Her-2/neu testing in breast cancer. Pathol Patterns Rev 2003, 120:53-71.
• [30]Sapino A, Coccorullo Z, Cassoni P, Ghisolfi G, Gugliotta P, Bongiovanni M, Arisio R, Crafa P, Bussolati G: Which breast carcinomas need Her2/neu gene study after immunohistochemical analysis? Results of combined use of antibodies against different c-erbB2 protein domains. Histopathology 2003, 43:354-362.
• [31]Cowell JK: Double minutes and homogeneously staining regions: gene amplification in mammalian cells. Annu Rev Genet 1982, 16:21-59.
• [32]Sauer T, Wiedswang G, Boudjema G, Chirstensen H, Karesen R: Assessment of HER-2/neu overexpression and/or gene amplification in breast carcinomas: should in situ hybridization be the method of choice? APMIS 2003, 111:444-450.
• [33]Sidoni A, Cavaliere A, Belleza G, Scheibel M, Bucciarelli E: Breast cancer in young women: clinicopathologial features and biological specificity. Breast 2003, 12:247-250.
• [34]Albain KS, Allred DC, Clark GM: Breast cancer outcome and predictors of outcome: are there age differentials? J Natl Cancer Inst Monogr 1994, 16:35-42.
• [35]Hoff ER, Tubbs RR, Myles JL, Procop GW: Her2/neu amplification in breast cancer: stratification of tumor type and grade. Am J Clin Pathol 2002, 117:916-921.