J Gynecol Oncol.  2009 Jun;20(2):113-116. 10.3802/jgo.2009.20.2.113.

TRAIL-induced cell death and caspase-8 activation are inhibited by cisplatin but not carboplatin

• Han SJ
• Ahn TK
• Choi HS
• Shin JN
• Piya S
• Kim TH

• KMID: 2245094
• DOI: http://doi.org/10.3802/jgo.2009.20.2.113

Abstract

OBJECTIVE
Platinum (Pt) based drugs including cisplatin and carboplatin are widely used as anticancer drugs in various human cancers. Many studies have shown that chemotherapeutic agents synergistically enhance cell death induced by death ligands. However it has been recently reported that cisplatin may inhibit tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-induced cell death through inactivation of caspases. Thus, we investigated whether carboplatin also inhibits TRAIL-induced cell death.
METHODS
HeLa cells were treated with TRAIL in the presence of cisplatin or carboplatin, and cell death was analyzed using the crystal violet staining method. Caspase activation was checked through detection of Bid cleavage by Western blotting using anti-Bid antibody.
RESULTS
Cisplatin inhibits TRAIL-induced cell death in HeLa cells; however, carboplatin enhanced TRAIL-induced cell death. Whereas cisplatin inhibited caspase-8-mediated Bid cleavage, carboplatin had no effect on caspase-8 activity.
CONCLUSION
Although cisplatin and carboplatin are platinum-containing cancer therapeutic agents, they have the opposite effects on TRAIL-induced cell death.

Keyword

Carboplatin; Caspase-8; Cisplatin; TRAIL

MeSH Terms

Blotting, Western
Carboplatin
Caspase 8
Caspases
Cell Death
Cisplatin
Gentian Violet
HeLa Cells
Humans
Ligands
Necrosis
Platinum
Carboplatin
Caspase 8
Caspases
Cisplatin
Gentian Violet
Ligands
Platinum

Figure

• 

  Fig. 1 Cisplatin inhibits TRAIL-induced cell death in HeLa cells, but not carboplatin. Cisplatin (100, 200 µg/ml) or carboplatin (100, 200 µg/ml) was incubated with/without TRAIL (100 ng/ml) for 24 hours in HeLa cells. (A) The cells were stained with Crystal violet. (B) Cell viability was measured by methanol extraction after crystal violet staining of the cells.

• 

  Fig. 2 Cisplatin directly inhibits activity of caspase-8, but not carboplatin. (A) Pretreated recombinant human caspase-8 (100 ng) with cisplatin (250 µg) was incubated with recombinant Bid (100 ng) in 50 µl of caspase assay buffer for 1 hour at 37℃. (B) Pretreated recombinant human caspase-8 (100 ng) with carboplatin (250 µg) in the presence or absence of pan-caspase inhibitor zVAD-FMK was incubated with recombinant Bid (100 ng) in 50 µl of caspase assay buffer for 1 hour at 37℃. Cleavage of Bid was analyzed by Western blot with anti-Bid antibody.

• 

  Fig. 3 Structure of cisplatin and carboplatin. Cisplatin contains a platinum atom complexed with two ammonia groups and two chlorides residue, and carboplatin contains a platinum atom complexed with two ammonia groups and 1,1-cyclobutane-dicarboxyl residue.

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