Correlation of cytotoxic effect of transmembrane and secretory TNF-α to cell cycle

Abstract

This study was aimed to examine the correlation of the cytotoxic effects induced by two types of TNF-α to cell cycle. Hoechst 33342 and PI were used to detect the morphological changes in the cell death induced by the two types of TNF-α. TdT and PI co-staining was performed to determine the phase of cell cycle of apoptotic cells. L929 cells in different phases of cell cycle were further synchronized and their sensitivity to the two types of TNF-α was observed. Our results showed that the apoptosis of HepG2 cells triggered by tm-TNF-α mainly occurred in G₁ phase while in HL-60, Raji and K562 cell lines it mainly took place in S phase. The apoptosis of L929 cells induced by tm-TNF-α mainly occurred in S phase while the apoptosis induced by s-TNF-α mainly appeared in G₁ phase. L929 cells were sensitive to s-TNF-α when synchronized in G₁ phase (cytotoxicity 49.8%) while their sensitivity to tm-TNF-α was highest in S phase (45.7%) and G₁/S phase (cytotoxicity 40.6%). It was concluded that tm-TNF-α-induced apoptosis of different target cells took place in different phases of cell cycle. The apoptosis of the specific cell line induced by the two types of TNF-α occurred in different phases of cell cycle. The sensitivity of the specific cell line to the two types of TNF-α was correlated with the phase of cell cycle.

Keywords

transmembrane TNF-α / secretory TNF-α / cytotoxicity / cell apoptosis / cell cycle

References

[1] WajantH., PfizenmaierK., ScheurichP.. Tumor necrosis factor signaling. Cell Death Differ, 2003, 10(1): 45-65

[2] HaranakaK., SatomiN.. Cytotoxic activity of tumor necrosis factor (TNF) on human cancer cells in vitro. Jpn J Exp Med, 1981, 51(3): 191-194

[3] MatthewsN., WatkinsJ.F.. Tumour-necrosis factor from the rabbit. I. Mode of action, specificity and physicochemical properties. Br J Cancer, 1978, 38(2): 302-309

[4] SugarmanB.J., AggarwalB.B., HassP.E., et al.. Recombinant human tumor necrosis factor-alpha: effects on proliferation of normal and transformed cells in vitro. Science, 1985, 230(4728): 943-945

[5] KrieglerM., PerezC., DeFayK., et al.. A novel form of TNF/cachectin is a cell surface cytotoxic transmembrane protein: ramifications for the complex physiology of TNF. Cell, 1988, 53(1): 45-53

[6] ShiW.F., LiZ.Y., GongF.L., et al.. Comparison of the cytocidal effect induced by transmembrane and secreted TNF-alpha. Chin J Microbiol Immunol^(Chinese), 1998, 18(6): 499-504

[7] KingK.L., CidlowskiJ.A.. Cell cycle regulation and apoptosis. Annu Rev Physiol, 1998, 60: 601-617

[8] ShihS.C., StutmanO.. Cell cycle-dependent tumor necrosis factor apoptosis. Cancer Res, 1996, 56(7): 1591-1598

[9] VermeulenK., BernemanZ.N., Van BockstaeleD.R.. Cell cycle and apoptosis. Cell Prolif, 2003, 36(3): 165-175

[10] LattimeE.C., StutmanO.. WEHI-164 clone 2F: in vitro antitumor effects of tumor necrosis factor and gamma-interferon. Nat Immun, 1992, 11(1): 34-45

[11] LattimeE.C., BykowskyM.J., StutmanO.. Susceptibility to lysis by natural killer and natural cytotoxic cells is independent of the mitotic stage of the target cell cycle. Cell Immunol, 1986, 100(1): 79-88

[12] SpyridopoulosI., MayerP., ShookK.S., et al.. Loss of cyclin A and G₁-cell cycle arrest are a prerequisite of ceramide-induced toxicity in human arterial endothelial cells. Cardiovasc Res, 2001, 50(1): 97-107

[13] VollandS., AmtmannE., SauerG.. TNF accelerates the S-phase of the cell cycle in tumor cells. Int J Cancer, 1994, 56(5): 698-705

[14] BharadwajU., Marin-MullerC., LiM., et al.. Mesothelin confers pancreatic cancer cell resistance to TNF-alpha-induced apoptosis through Akt/PI3K/NFkappaB activation and IL-6/Mcl-1 overexpression. Mol Cancer, 2011, 10: 106

[15] FaracoP.R., LedgerwoodE.C., VandenabeeleP., et al.. Tumor necrosis factor induces distinct patterns of caspase activation in WEHI-164 cells associated with apoptosis or necrosis depending on cell cycle stage. Biochem Biophys Res Commun, 1999, 261(2): 385-392

[16] YangL., ShiW., WangJ., et al.. Comparison of cell deaths induced by transmembrane and secretory TNF-alpha. J Huazhong Univ Sci Technol [Med Sci], 2007, 27(2): 117-119