The anticancer effects of
kotomolide A (KTA), a new butanolide constituent isolated from the leaves of Cinnamomum kotoense (Lauraceae), on the two human
breast cancer cell lines MCF-7 and MDA-MB-231, were first investigated in our study. KTA exhibited selectively antiproliferative effects in
cancer cell lines without showing any toxicity in normal mammary epithelial cells. Treatment of
cancer cells with KTA to trigger G2/M phase arrest was associated with increased p21/WAF1 levels and reduced amounts of
cyclin A,
cyclin B1, cdc2 and cdc25C. KTA induced
cancer cell death treatment by triggering mitochondrial and
death receptor 5 (DR5) apoptotic pathways, but did not act on the
Fas receptor. Exposure of MCF-7 and MDA-MB-231 cells to KTA resulted in cellular
glutathione reduction and ROS generation, accompanied by JNK activation and apoptosis. Both
antioxidants, NAC and
catalase, significantly decreased apoptosis by inhibiting the phosphorylation of JNK and subsequently triggering DR5 cell death pathways. The reduction of JNK expression by
siRNA decreased KTA-mediated Bim cleavage, DR5 upregulation and apoptosis. Furthermore, daily KTA i.p.
injections in nude mice with MDA-MB-231 s.c.
tumors resulted in a 50% decrease of mean
tumor volume, compared with vehicle-treated controls. Taken together, the data show that cell death of
breast cancer cells in response to KTA is dependent upon ROS generation and JNK activation, triggering intrinsic and extrinsic apoptotic pathways. The ROS/JNK pathway could be a useful target for novel approaches in
breast cancer chemotherapy.