Expression and location of pro-apoptotic bcl-2 family protein BAD in normal human tissues and tumor cell lines

Shinichi Kitada, Maryla Krajewska, Xin Zhang, Dominic Scudiero, Juan M. Zapata, Hong Gang Wang, Ahmed Shabaik, Gabriella Tudor, Stanislaw Krajewski, Timothy G. Myers, George S. Johnson, Edward A. Sausville, John C. Reed

Research output: Contribution to journalArticle

102 Scopus citations

Abstract

The BAD protein is a pro-apoptotic member of the Bcl-2 family whose ability to heterodimerize with survival proteins such as Bcl-X(L) and to promote cell death is inhibited by phosphorylation. Monoclonal antibodies were generated against the human BAD protein and used to evaluate its expression by immunoblotting and immunohistochemistry in normal human tissues and by immunoblot analysis of the National Cancer Institute anti-cancer drug screening panel of 60 human tumor cell lines. BAD protein was detectable by immunoblotting in many normal tissues, with testis, breast, colon, and spleen being among those with the highest steady-state levels. Immunostaining of tissues revealed many examples of cell-type-specific expression of BAD, suggesting dynamic regulation of BAD protein levels in vivo. In many types of normal cells, BAD immunoreactivity was associated with cytosolic organelles resembling mitochondria, suggesting that BAD is often heterodimerized with other Bcl-2 family proteins in vivo. The relative levels of BAD protein varied widely among established human tumor cell lines, with colon, lung, and melanomas generally having the highest expression. As a group, hematopoietic and lymphoid lines contained the least BAD protein. The BAD protein derived from 11 of 41 tumor lines that expressed this pro-apoptotic protein migrated in gels as a clear doublet, consistent with the presence of hyperphosphorylated BAD protein. Taken together, these findings define for the first time the normal cell-type-specific patterns of expression and intracellular locations of the BAD protein in vivo and provide insights into the regulation of this pro-apoptotic Bcl-2 family protein in human tumors.

Original languageEnglish (US)
Pages (from-to)51-61
Number of pages11
JournalAmerican Journal of Pathology
Volume152
Issue number1
StatePublished - Jan 1 1998

All Science Journal Classification (ASJC) codes

  • Pathology and Forensic Medicine

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    Kitada, S., Krajewska, M., Zhang, X., Scudiero, D., Zapata, J. M., Wang, H. G., Shabaik, A., Tudor, G., Krajewski, S., Myers, T. G., Johnson, G. S., Sausville, E. A., & Reed, J. C. (1998). Expression and location of pro-apoptotic bcl-2 family protein BAD in normal human tissues and tumor cell lines. American Journal of Pathology, 152(1), 51-61.