Toona sinensis and Moschus are two herb materials used in traditional Chinese medicine, most commonly for their various biological activities. In this study, we investigated the inhibitory effect of three decoctions from Toona sinensis, Moschus, and Toona sinensis and Moschus in combination on cell growth in several normal and cancer cell lines by cell viability assay. The results showed that the combined decoction exhibited the strongest anticancer effects, compared to two single decoctions. The observations indicated that the combined decoction did not induce cell apoptosis and autophagy in HeLa cells by fluorescence microscopy. Flow cytometry analysis revealed that the combined decoction arrested HeLa cell cycle progression in S-phase. After the decoction incubation, among 41 cell cycle related genes, eight were reduced, while five were increased in mRNA levels by real-time PCR assay. Western blotting showed that there were no apparent changes of protein levels of Cyclin E1, while P27 expression significantly declined and the levels of CDC7 and CDK7 obviously increased. The data suggest that the RB pathway is partially responsible for the decoction-induced S-phase cell cycle arrest in HeLa cells. Therefore, the combined decoction may have therapeutic potential as an anticancer formula for certain cancers. 1. Introduction Traditional Chinese medicine (TCM) has been used in clinical practice for thousand years. Compound formula of TCM has been shown to exhibit synergism [1]. TCMs are used to restore overall healthful balance and normal body function in a holistic way due to their moderate treatment effects and lower side effects [2]. These features made themselves popular in China. Toona sinensis is a type of arbor that is widely distributed in Asia. The leaves and young shoots are used as vegetable in China and Malaysia [3, 4]. In fact, it has long been used in TCM for a wide variety of conditions. The leaf extracts showed various biological activities, including anticancer [5–9], antidiabetes [10], and antioxidant [11] effects, as well as inhibiting Leydig cell steroidogenesis [12] and suppressing brain degeneration in senescence-accelerated mice [13]. The bark is used as astringent and depurative, the powdered root is used as a corrective, and the fruits are used as an astringent and for the treatment of eye infection [14, 15]. Musk, a ventral glandular secretion of the male musk deer, is also a precious and wide applied material in TCM [16–18]. As a major Chinese herbal material, musk was firstly recorded in Shen Nong Ben Cao Jing (The Herbal
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