Protein studies of human non‐Hodgkin's B‐lymphoma: Appraisal by two‐dimensional gel electrophoresis

Abstract

We have utilized two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) coupled with silver stain to identify cellular proteins in human non-Hodgkin's B-lymphoma (NHL). Five cell lines (SKDHL2B, WSU-DLCL2, WSU-NHL, WSU-FSCCL and SKLN1), representing four different NHL maturational stages and a normal Epstein-Barr virus (EBV)-transformed line of B-cell origin (SKLN1) were studied. The NHL lines were immunophenotyped using flow cytometry with lineage associated monoclonal antibodies. Whole cell lysates of the cell lines were subjected to 2-D PAGE analyses. The gels were analyzed with an image scanning computer and the qualitative differences of protein patterns were studied. Results revealed great similarities in patterns of the NHL lines. A master map containing common NHL-protein spots was constructed. When the map of each tumor line was compared to the master map, several protein spots were associated with each NHL-grade. Search for these proteins in the normal EBV-transformed B-cell line showed that only one of the proteins (S3; M(r)/pI 19/5.9) was present. Proteins that were detected in malignant NHL, but not in the normal EBV-line, could provide important information regarding the human NHL B-lymphocyte data-bases. Whether or not these proteins are definite malignant markers to distinguish between different NHL maturational stages needs further exploration through electroblotting and microsequencing.

2‐D PAGE‐based comparison of proteasome inhibitor bortezomib in sensitive and resistant mantle cell lymphoma

“…However, results of cell line experiments may be transferred to primary disease only with caution as additional “bystander” alterations unrelated to the MCL pathogenesis might have accumulated in single cell lines during establishment. Using a panel of MCL cell lines with various secondary alterations in a pooled analysis and verifying the candidate proteins in primary MCL cells allowed to identify the critical effects of proteasome inhibition 16, 17.…”