Detection of bcl-2 protein and bcl-2 messenger RNA in normal and neoplastic lymphoid tissues by immunohistochemistry and in situ hybridization

Abstract

bcl-2 protein has been detected in surgical specimens and cultured permanent cell lines of non-Hodgkin's lymphomas and leukemias using enzyme immunohistochemistry and immunofluorescence with anti-bcl-2 monoclonal antibodies. Of 40 surgical specimens, bcl-2 protein was expressed in 50% of B-cell and 41% of T-cell lymphomas, both with and without the bcl-2 gene rearrangement. In investigations of 38 hematopoietic cell lines, bcl-2 protein was detected not only in lymphoid cell lines but also in myeloid cell lines. In situ hybridization and immunohistochemical analysis of reactive lymph nodes showed that lymphocytes in mantle zones and paracortical areas expressed bcl-2 protein consistent with the messenger RNA distribution and that germinal center cells showed abundant bcl-2 transcript, despite the absence of detectable bcl-2 protein. These results suggest that bcl-2 protein is broadly expressed in various hematopoietic neoplasms not restricted in t(14; 18) lymphomas and that germinal center cells may be involved in some arrest of bcl-2 protein expression at the posttranscriptional level.

Immunoreactivity for bcl‐2 protein in cutaneous lymphomas and lymphoid hyperplasias*

“…However, intense labelling for this protein was also demonstrable in small lymphoeytes and imtiiunoblastoid eells in ,33% of CLH eases, and it was seemingly independent of lymphoeytic litieage in that cotitext. This tesitlt is in keeping with the observation that BCLP is present in mature B and T lymphoeytes of the mantle zoties and intetfoUicular regiotis of hyperplastic lymph nodes (6,7). These data indicate that imnitmostains for BCLP are of little help in the general distitietion of MLS from CLH, without speeific segregation into diffuse and foUienlar proliferations.…”

“…25). Although it was initially thought to be restrieted to lymphomas bearing the (14; 18) translocatioti (9), BCLP has subsequently been demonstrated in a broad spectrum of non-Hodgkin's lymphomas with both B-atid T-cell phetiotypes as well as some examples of Hodgkin's disease (5)(6)(7)27).…”

“…This obsei-vation suggests that the frequency of BC^LP-positivit)' in lymphomas of the skin is influeneecl by the histologieal tumor type, as it is in extraeutaneovts tumors. The latter fact will be an important variable to consider in cotiiparative analyses of ftttute studies of BCCLP in MLS (6,7).…”

Flow cytometric analysis of apoptosis and BCL-2 in primary breast carcinomas: Clinical and biological implications

“…These studies have largely been conducted by immunohistochemical and/or molecular techniques. Flow cytometric applications are relatively few and have primarily been applied to hematoreticular malignancies ( 1,5,8,23,33,46). Similar investigations in solid tumors are rare (14,49).…”

Germinal center‐associated nuclear protein (GANP) is involved in mRNA export of Shugoshin‐1 required for centromere cohesion and in sister‐chromatid exchange

“…Digoxigenin-labeled anti-sense and the sense (control) probes were generated using an RNA labeling in vitro transcription kit (Roche) and hSgo1 cDNA (331-757 nt) in the coding region in the pGEM-T vector (Promega). The hBcl2 probe was prepared (Kondo et al 1992). siRNA-treated cells were fixed with 4% paraformaldehyde ⁄ PBS, followed by proteinase K treatment and acetylation, and then hybridized with the probe for 16 h at 50°C.…”