-Hydroxyisovalerylshikonin Inhibits the Cell Growth of Various Cancer Cell Lines and Induces Apoptosis in Leukemia HL-60 Cells through a Mechanism Different from Those of Fas and Etoposide

Abstract

beta-Hydroxyisovalerylshikonin (beta-HIVS), which was isolated from the plant, Lithospermium radix, inhibited the growth of various lines of cancer cells derived from human solid tumors at low concentrations between 10(-8) and 10(-6) M. When HL-60 cells were treated with 10(-6) M beta-HIVS for 3 h, characteristic features of apoptosis, such as DNA fragmentation, nuclear fragmentation, and activation of caspase-3-like activity, were observed. The most characteristic features of the effect of beta-HIVS were the remarkable morphological changes induced upon treatment of HL-60 cells with beta-HIVS, as visualized on the staining of actin filaments with phalloidin labeled with tetramethylrhodamine B isothiocyanate. Moreover, activation of MAP kinases, such as ERK2, JNK and p38, was detected after treatment with 10(-6) M beta-HIVS preceding the appearance of the characteristics of apoptosis, and the features of the activation of these MAP kinases were quite different from those of Fas and anticancer drug-induced apoptosis. The activation of JNK by beta-HIVS was not inhibited by inhibitors of caspases, suggesting that JNK is located either upstream or independent of the caspase signaling pathway. beta-HIVS did not inhibit the activity of topoisomerase II. These results indicate that beta-HIVS induces apoptosis in HL-60 cells through a mechanism unlike those reported for anti-Fas antibodies and etoposide.

Apoptosis induced by arsenic trioxide in leukemia U937 cells is dependent on activation of p38, inactivation of ERK and the Ca2+-dependent production of superoxide

“…␤-Hydroxyisovalerylshikonin (␤-HIVS) was purified from the plant Lithospermium radix as described previously. 18 Dihydroethydium, also called hydroethydine (DE) and 2Ј,7Ј-dichlorodihydrofluorescein diacetate, also called 2Ј,7Ј-dichlorofluoresin diacetate (DCFH-DA), were from Molecular Probes (Eugenes, OR). PD98059 and SB203580 were obtained from New England BioLabs (Beverly, MA) and Calbiochem (LaJolla, CA), respectively.…”

“…18,21 In brief, JNK was precipitated from an aliquot of cell lysate that contained 400 g of protein by mixing the lysate with recombinant GST-c-JUN(1-79) that had been conjugated to glutathione-Sepharose. The Sepharose beads were pelleted by brief centrifugation in a microcentrifuge, and the bound protein complexes were washed three times with N-(2-hydroxyethyl)piperazine-NЈ-2-ethane-sulfonic acid (HEPES) binding buffer (20 mM HEPES, pH 8.0, 20 mM MgCl 2 , 0.1 mM EDTA, 50 mM NaCl, 0.05% Triton X-100).…”

β-Hydroxyisovalerylshikonin induces apoptosis in human leukemia cells by inhibiting the activity of a polo-like kinase 1 (PLK1)

“…The importance of apoptosis has been emphasized by recent demonstrations that various chemotherapeutic anti-cancer agents, such as cisplatin (Kaufmann, 1989;Barry et al, 1990;Evans and Dive, 1993), adriamycin (Friesen et al, 1996), and taxol (Bhalla et al, 1993), induce apoptosis in cancer cells. We have identified several differentiationor apoptosis-inducing compounds (Sakai et al, 1993(Sakai et al, , 1994Ohizumi et al, 1995;Masuda et al, 1995Masuda et al, , 1997Watabe et al, 1996) and found recently that bhydroxyisovalerylshikonin (b-HIVS) has potent apoptosis-inducing activity (Hashimoto et al, 1999). We isolated b-HIVS from a Chinese plant, Lithospermum erythrorhizon, and found that it had the strongest apoptosis-inducing activity of a variety of derivatives of shikonin in HL60 cells (Hashimoto et al, 1999).…”

“…We have identified several differentiationor apoptosis-inducing compounds (Sakai et al, 1993(Sakai et al, , 1994Ohizumi et al, 1995;Masuda et al, 1995Masuda et al, , 1997Watabe et al, 1996) and found recently that bhydroxyisovalerylshikonin (b-HIVS) has potent apoptosis-inducing activity (Hashimoto et al, 1999). We isolated b-HIVS from a Chinese plant, Lithospermum erythrorhizon, and found that it had the strongest apoptosis-inducing activity of a variety of derivatives of shikonin in HL60 cells (Hashimoto et al, 1999). In the present study, we tried to identify the gene that is involved in b-HIVS-induced apoptosis to clarify the molecular mechanisms of apoptosis.…”

Lithospermi radixExtract Inhibits Histamine Release and Production of Inflammatory Cytokine in Mast Cells

“…37,38) Shikonin has been reported to confer many medicinal properties such as anti-inflammatory and anti-tumor effects. 2,39) we therefore assumed that the effects of LR in this study would be related to shikonin or its derivatives.…”