A Comparison of Apoptosis and Necrosis Induced by ent-Kaurene-Type Diterpenoids in HL-60 Cells

Abstract

We previously reported that ent-11α-hydroxy-16-kauren-15-one (KD) induces apoptosis through a caspase-dependent pathway and the induction of apoptosis is dependent on its enone group in human leukemia cells. Here we investigated the abilities of some KD-related compounds with enone group (Fig. [1]) to induce apoptosis and to activate some caspases. The IC₅₀ values of ent-kaurene-related compounds possessing the enone group, ent-1β-hydroxy-9(11),16-kauradien-15-one (1), ent-9(11),16-kauradiene-12,15-dione (2) and the rearranged ent-kaurane-type diterpene (3), against HL-60 cells after 12 h of treatment were 40 μM, 1.8 μM and 5.5 μM, respectively. Although treatment with 3 induced apoptosis, DNA ladder formation was not observed after treatment with 1 or 2. Induction of necrosis, as assayed by trypan blue staining, was observed after treatment with 1 or 2. Treatment with compound 1, 2 or 3 induced proteolysis of poly(ADP-ribose) polymerase (PARP), a substrate of caspase-3, and processing of caspase-3. Activation of caspase-8 and processing of Bid, a typical substrate of caspase-8, were also observed on treatment with these compounds. Pretreatment with a broad-spectrum inhibitor of caspases attenuated apoptosis induced by 3 but not necrosis induced by 1 and 2. In summary, KD-related compounds are a unique family of diterpenes that cause either caspase-dependent apoptotic or necrotic cell death.

Abbreviations

KD:ent-11α-hydroxy-16-kauren-15-one

PARP:poly(ADP-ribose) polymerase

NF-κB:nuclear factor-κB

Key words

Kaurene - apoptosis - caspase - HL-60 cell - Jungermannia truncata - Jungermanniales

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