Evidence for a protein synthesis-dependent and -independent TNF alpha cytolytic mechanism

Gynecologic Oncology
C B PowellJ L Collins

Abstract

The analysis of the effect of the protein synthesis inhibitors emetine (EM) or actinomycin D (ACT-D) on the TNF alpha-mediated cytolysis of L929 target cells demonstrates a biphasic, concentration (10(-12)-10(-4) M)-dependent curve indicative of two cytolytic mechanisms operative in L929 cells. One TNF alpha cytolytic mechanism is dependent on protein synthesis in the target cells, while the other cytolytic mechanism is protein synthesis independent. Both TNF alpha cytolytic mechanisms cause apoptosis (fragmentation of DNA) as shown by the TNF alpha-mediated release of tritiated thymidine, Apoptag, and DAPI staining, in the presence or absence of EM or ACT-D. The two cytolytic mechanisms are also similar in their requirement for lipoxygenase enzymes as shown by the ability of nordihydroguaiaretic acid (10(-6)-10(-5) M) and ketoconazole (4 x 10(-6)-2 x 10(-5) M) to block TNF alpha-mediated lysis of the target cells. However, the two cytolytic mechanisms differ in their requirement for the production of oxygen free radicals. The oxygen free radical scavengers, dimethylsulphoxide (0.2-0.4 M) and glutathione (2 x 10(-6)-10(-5) M) block the TNF alpha-mediated cytolysis of target cells in the absence of protein synthesis inhibitors, ...Continue Reading

Citations

Jul 27, 1999·Clinical Chemistry and Laboratory Medicine : CCLM·M GräfeE Fleck
Aug 16, 2001·Critical Reviews in Toxicology·E J Calabrese
Aug 1, 1997·Immunological Investigations·M StangelP Marx

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