Immunohistochemical Profiling of Lymphoma

Representative areas with the highest percentage of tumor cells were selected for tissue microarray (TMA) construction as described previously (5 (link)). Immunohistochemical (IHC) studies for various markers were performed; B-cell lymphoma 2 (BCL2), B-cell lymphoma 6 (BCL6), cyclin D1, CD10, CD30, Forkhead box protein P1 (FOXP1), Germinal Center B cell-expressed Transcript-1 (GCET1), Multiple Myeloma Oncogene 1 (MUM1), MYC, Nuclear factor-κB (NF-κB) components (p50, p52, p65, and c-Rel), p53, phosphorylated protein kinase B (pAKT), and phosphorylated signal transducer and activator of transcription 3 (pSTAT3). Receiver-operating characteristic (ROC) curves and X-tile analyses were utilized to assess a cutoff (5 (link)). When an optimal cutoff could not be determined by ROC curve and X-tile analyses, a conventional cutoff value for individual markers was decided based on previous reports in the literature. The cutoff scores for these markers were as follows: 10% for cyclin D1; 20% for CD30 and p53; 30% for CD10, and BCL6, 40% for MYC; 50% for pSTAT3; 60% for GCET1, MUM1 and FOXP1; and 70% for AKT and BCL2. All markers except cyclin D1 were determined by ROC curve. Any nuclear expression of each NF-κB component was considered positive.

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Ok C.Y., Chen J., Xu-Monette Z.Y., Tzankov A., Manyam G.C., Li L., Visco C., Montes-Moreno S., Dybkær K., Chiu A., Orazi A., Zu Y., Bhagat G., Richards K.L., Hsi E.D., Choi W.W., Han van Krieken J., Huh J., Zhao X., Ponzoni M., Ferreri A.J., Bertoni F., Farnen J.P., Møller M.B., Piris M.A., Winter J.N., Medeiros L.J, & Young K.H. (2014). Clinical Implications of Phosphorylated STAT3 Expression in de novo Diffuse Large B-cell Lymphoma. Clinical cancer research : an official journal of the American Association for Cancer Research, 20(19), 5113-5123.

Publication 2014

B cell B cell lymphoma Bcl6 C rel Cells Cyclin d1 Forkhead box protein Germinal center Microarray Multiple myeloma oncogene 1 Nuclear factor κb Protein kinase b Signal transducer and activator of transcription 3 Tissue Tumor

Corresponding Organization : The University of Texas MD Anderson Cancer Center

Other organizations : Taizhou University, University Hospital of Basel, Ospedale San Bortolo, Marqués de Valdecilla University Hospital, Aalborg University Hospital, Kettering University, Memorial Sloan Kettering Cancer Center, Cornell University, Methodist Hospital, NewYork–Presbyterian Hospital, Columbia University Irving Medical Center, University of North Carolina at Chapel Hill, Cleveland Clinic, University of Hong Kong, Radboud University Nijmegen, Radboud University Medical Center, Asan Medical Center, Second Affiliated Hospital of Zhejiang University, San Raffaele University of Rome, Istituto Oncologico della Svizzera Italiana, Lutheran Hospital, Odense University Hospital, Northwestern University

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Variable analysis

independent variables

Representative areas with the highest percentage of tumor cells

dependent variables

Expression levels of various markers: BCL2, BCL6, cyclin D1, CD10, CD30, FOXP1, GCET1, MUM1, MYC, NF-κB components (p50, p52, p65, and c-Rel), p53, pAKT, and pSTAT3

control variables

Not explicitly mentioned

controls

Receiver-operating characteristic (ROC) curves and X-tile analyses were utilized to assess a cutoff for the marker expression levels.
When an optimal cutoff could not be determined by ROC curve and X-tile analyses, a conventional cutoff value for individual markers was decided based on previous reports in the literature.

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