Multiple pathways were involved in tubeimoside-1-induced cytotoxicity of HeLa cells.

The Bolbostemma paniculatum (Maxim.) Franquet (Cucurbitaceae) is a Chinese herb with anticancer potential. Its main active component tubeimoside-1 (TBMS1), a triterpenoid saponin, was previously proved as a potent anticancer chemotherapeutic agent; however, the molecular basis for its activities is still elusive. In the present study, subcellular proteomic study in the cytoplasm and membrane protein fractions extracted from HeLa cells revealed that proteins act as mediators of ROS generation and Ca(2+) regulation were substantially altered in expression upon TBMS1 stimuli. We also found that TBMS1 induced cell cycle arrest at G2/M phase accompanied by a decrease in G0/G1 phase in HeLa cells. Further biochemical studies showed that TBMS1 inhibited the levels of cyclinB1, Cdc2 and Cdc25C, but enhanced Chk2 phosphorylation. In addition, the cytoplasm sequestration of Cdc25C, Cip1/p21 induction and tubulin dyspolymerization also contributed to the TBMS1-mediated cell cycle arrest on the G2/M phase.