Berberine Induces Apoptotic Cell Death via Activation of Caspase-3 and -8 in HL-60 Human Leukemia Cells: Nuclear Localization and Structure–Activity Relationships

Abstract

Berberine (BBR), an isoquinoline alkaloid, is a well-known bioactive compound contained in medicinal plants used in traditional and folk medicines. In this study, we investigated the subcellular localization and the apoptotic mechanisms of BBR were elucidated. First, we confirmed the incorporation of BBR into the cell visually. BBR showed antiproliferative activity and promptly localized to the nucleus from 5 min to 15 min after BBR treatment in HL-60 human promyelocytic leukemia cells. Next, we examined the antiproliferative activity of BBR (1) and its biosynthetically related compounds (2-7) in HL-60 cells. BBR exerted strongest antiproliferative activity among 1-7 and the results of structures and activity relation suggested that a methylenedioxyl group in ring A, an O-alkyl group at C-9 position, and the frame of isoquinoline may be necessary for antiproliferative activity. Moreover, BBR showed the most potent antiproliferative activity in HL-60 cells among human cancer and normal cell lines tested. Next, we examined the effect of BBR on molecular events known as apoptosis induction. In HL-60 cells, BBR induced chromatin condensation and DNA fragmentation, and triggered the activation of PARP, caspase-3 and caspase-8 without the activation of caspase-9. BBR-induced DNA fragmentation was abolished by pretreatment with inhibitors against caspase-3 and caspase-8, but not against caspase-9. ERK and p38 were promptly phosphorylated after 15 min of BBR treatment, and this was correlated with time of localization to the nucleus of BBR. These resultsCorrespondence to: Prof. Yukihiro Shoyama, Department of Pharmacognosy, Faculty of Pharmaceutical Sciences, Nagasaki International University, 2825-7 Huis Ten Bosch, Sasebo, Nagasaki 859-3298, Japan. Tel: (þ81) The American Journal of Chinese Medicine, Vol. 45, No. 7, 1497-1511 demonstrated that BBR translocated into nucleus immediately after treatments and induced apoptotic cell death by activation of caspase-3 and caspase-8.

Potential suppressive effects of theophylline on human rectal cancer SW480 cells in�vitro by inhibiting YKL-40 expression

“…Therefore modulating YKL-40 functions may represent an efficient strategy for rectal cancer prevention and treatment. In recent years, certain compounds extracted from traditional herbal medicines and plants have indicated potential at preventing and curing tumors (25). Theophylline is the main constituent and characteristic component of the purine alkaloids found in tea plants; it is also a specific inhibitor of YKL-40 (26).…”

Berberine induces mitochondrial‑mediated apoptosis and protective autophagy in human malignant pleural mesothelioma NCI‑H2452 cells

“…Berberine has extensive pharmacological effects, including anti-hypertensive, anti-diabetic, anti-arrhythmia and anti-hyperlipidemic (3)(4)(5)(6). Recent studies have shown that berberine exerts antitumor effects in a variety of human cancer cells, such as breast cancer, hepatocellular carcinoma, leukemia, and esophageal cancer, both in vitro and in vivo, by suppressing proliferation, inducing apoptosis, arresting the cell cycle, and inhibiting invasion and metastasis (7)(8)(9)(10)(11)(12). In addition, it has been recently reported that berberine-induced autophagy has been observed in hepatic and colon cancer cells (13,14).…”

Self-nanoemulsifying system improves oral absorption and enhances anti-acute myeloid leukemia activity of berberine

“…It has many pharmacological actions, including activity against tumors [ 13 , 14 ], microbe [ 15 ], inflammation [ 15 , 16 ], neurodegenerative disease [ 17 , 18 ] and other diseases. Recent studies have found that BBR exerts cytotoxicity and inhibits telomerase and topoisomerase, promotes the proliferation apoptosis in human leukemia cells by specifically binding to oligonucleotides or polymorphic nucleic acid and by stabilizing DNA triplexes or G-quadruplexes; the electrostatic interactions may be quantified in terms of the Hill model of cooperative interactions [ 19 – 22 ]. BBR showed antiproliferative activity and promptly localized to the nucleus 5 min to 15 min after BBR treatment in HL-60 human promyelocytic leukemia cells [ 22 ].…”