Cis-bixin has antineoplastic effects in multiple cancer cell lines. (A) Photo of seed pods of the rain forest plant Bixa orellena, or annatto (Alan J. Wolf, Ph.D. ©2006, University of Wisconsin-Madison). (B) Chemical structures of cis-bixin and several related carotenoids. (C) Colony-forming assay for cytotoxicity using A549 cells shows that cis-bixin is more cytotoxic than are tested related carotenoids. (D) Colony-forming assays in a variety of cancer cell lines, including A549 (lung), U2OS (osteosarcoma), PC3 (prostate), HCT-118 (colon), MCF-7 (breast), DRO (anaplastic thyroid), and BHP5-16 (papillary thyroid) indicate broad anticancer effects of cis-bixin. All drug exposures were for 24 h. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article at www.liebertonline.com/ars).