Bcl-2 modulates Fas-mediated apoptosis in human renal cell carcinoma cell lines.
The objective of this study was to characterize the role of Bcl-2 expression in Fas-mediated apoptosis in human renal cell carcinoma (RCC) cell lines. RT-PCR analyses showed that 10 different RCC cell lines expressed Fas, but not Fas ligand. Seven of 10 cell lines expressed Fas strongly, while 3 cell lines weakly expressed Fas by flow cytometric analyses. Measurement of the LDH concentration in the culture supernatant revealed that 6 of the 7 cell lines which expressed Fas strongly were sensitive to treatment with an agonistic anti-Fas monoclonal antibody (CH11), whereas all cell lines which weakly expressed Fas did not show Fas-mediated cell death. Furthermore, Bcl-2 expression was found only in three cell lines which were all susceptible to CH11, and the downregulation of Bcl-2 protein by treatment with antisense oligodeoxynucleotide targeting Bcl-2 gene resulted in an enhancement of cell death induced by CH11. These findings suggest that strong Fas expression is necessary for Fas-mediated cell death in RCC cell lines, and Bcl-2 has a protective role against treatment with an anti-Fas monoclonal antibody, despite the fact that Bcl-2 expression was observed only in sensitive cell lines to Fas-mediated cell death.
Hara
I
I
Department of Urology, Kobe University School of Medicine, Kobe 650-0017, Japan. hara@med.kobe-u.ac.jp
Hara
S
S
Miyake
H
H
Arakawa
S
S
Kamidono
S
S
D000911:Antibodies, Monoclonal; D000937:Antigen-Antibody Reactions; D017209:Apoptosis; D015153:Blotting, Western; D002292:Carcinoma, Renal Cell / Q000378:metabolism* / Q000473:pathology; D002455:Cell Division; D053938:DNA Fragmentation; D017931:DNA Primers / Q000737:chemistry; D005434:Flow Cytometry; D006801:Humans; D007680:Kidney Neoplasms / Q000378:metabolism* / Q000473:pathology; D020320:Oligoribonucleotides, Antisense / Q000494:pharmacology; D011518:Proto-Oncogene Proteins / Q000378:metabolism; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction; D014162:Transfection; D014407:Tumor Cells, Cultured; D051028:bcl-2-Associated X Protein; D019014:fas Receptor / Q000235:genetics / Q000276:immunology / Q000502:physiology*
D000911:Antibodies, Monoclonal; D017931:DNA Primers; D020320:Oligoribonucleotides, Antisense; D011518:Proto-Oncogene Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D051028:bcl-2-Associated X Protein; D019014:fas Receptor