Functional proteomic and structural insights into molecular targets related to the growth inhibitory effect of tanshinone IIA on HeLa cells

Abstract

Certain antitumor agents have recently been extracted from the roots of Salvia miltiorrhiza Bunge. The diterpene derivative, tanshinone IIA, possesses cytotoxic activity against several human carcinoma cell lines. It also inhibits invasion and metastasis of cancer cells. In the present study, we isolated tanshinone IIA from S. miltiorrhiza, and it exhibited strong growth inhibition against human cervical cancer cells in dose- and time-dependent manners with a 50% cell growth inhibition value of 2.5 microg/mL (8.49 microM). Flow cytometric analysis of cell cycle progression revealed that G(2)/M arrest was initiated after a 24 h exposure to the drug. It also resulted in DNA fragmentation and degradation of poly (ADP-ribose) polymerase indicating that tanshinone IIA may be a potential antitumor agent. Furthermore, we performed a comprehensive proteomic analysis to survey global protein changes induced by tanshinone IIA treatment on HeLa cells. Significant changes in the levels of cytoskeleton proteins as well as stress-associated proteins were observed. Immunoblot analysis and immunofluorescence staining were used to confirm the levels of protein expression. Overexpression of the vimentin rescued these tanshinone IIA-induced events. Computational docking methods indicated that tanshinone IIA could stably bind to the beta-subunit of the microtubule protein. An interaction network analysis of these 12 proteins using MetaCore software suggested that tanshinone IIA treatment regulated the expressions of proteins involved in apoptotic processes, spindle assembly, and p53 activation, including vimentin, Maspin, alpha- and beta-tubulin, and GRP75. Taken together, our results suggest that tanshinone IIA strongly inhibited the growth of cervical cancer cells through interfering in the process of microtubule assembly, leading to G(2)/M phase arrest and sequent apoptosis. The success of this large-scale effort was assessed by a bioinformatics analysis of proteins through predictions of protein domains and possible functional roles. The possible contributions of these proteins to the cytotoxicity of tanshinone IIA provide potential opportunities for the development of cancer therapeutics.

Inhibitory effects of Scutellaria baicalensis extract on hepatic stellate cells through inducing G2/M cell cycle arrest and activating ERK-dependent apoptosis via Bax and caspase pathway

“…The effects of SBEE on cell cycle were evaluated by flow cytometry (Pan et al, 2010). HSC-T6 cells were treated with SBEE with the indicated concentrations for 24 h and harvested by centrifugation at 750 × g for 5 min at room temperature.…”

Tanshinone IIA Reverses the Malignant Phenotype of SGC7901 Gastric Cancer Cells

“…Recent researches have shown that TIIA is a natural anti-tumor drug, which has cytotoxic effect on many tumors and inhibits the invasion and migration of cancer cells. For example, TIIA could inhibit the cell growth of reproductive system tumors, such as breast cancer (Su and Lin, 2008;Lu et al, 2009), cervical cancer (Pan et al, 2010), ovarian cancer (Jiao and Wen, 2011) and prostatic carcinoma (Won et al, 2010); TIIA disrupts the membrane potential of mitochondria, induces apoptosis and reduces the adhesion and invasion of cells in acute promyelocytic leukemia. However, few studies have been reported to investigate the effect of TIIA on gastric cancer.…”

Effects of some elicitors on tanshinone production in adventitious root cultures of Perovskia abrotanoides Karel

“…Tanshinones, abietane-type norditerpenoid quinones, have been shown to exhibit diverse pharmacological activities, including antibacterial (Lee et al, 1999), antioxidant (Cao et al, 1996), antidiabetes (Kim et al, 2007), anti-cancer (Wang et al, 2005; 2009; Chiu and Su, 2010;Pan et al, 2010) and anti-inflammatory (Jang et al, 2003) activity. Since the roots of P. abrotanoides contain therapeutically applicable tanshinones, the mass cultivation of roots in vitro could be an effective technique for the large-scale production of these valuable secondary metabolites.…”