Abstract
BackgroundArdisia pusilla A. DC., family Myrsinaceae, is a traditional Chinese medicine named Jiu Jie Long with a variety of pharmacological functions including anti-cancer activities. In this study, we purified a natural triterpenoid saponin, ardipusilloside I, from Ardisia pusilla, and show that it exhibits inhibitory activities in human mucoepidermoid carcinoma Mc3 cells. We also investigated the underlying mechanisms of proliferation inhibition that ardipusilloside I exerts on Mc3 cells.MethodsMTT test was used to detect cell proliferation. Cell apoptosis was detected by transmission electron microscopy, Hoechst-33342 staining, DNA fragmentation detection, and flow cytometry. We also used western blot analysis to detect the potential mechanisms of apoptosis.ResultsArdipusilloside I affected the viability of Mc3 cells in a dose- and time-dependent manner. The IC50 of ardipusilloside I was approximately 9.98 μg/ml at 48 h of treatment. Characteristic morphological changes of apoptosis, including nuclear condensation, boundary aggregation and splitting, and DNA fragmentation, were seen after treatment with 10 μg/ml ardipusilloside I for 48 h. Western blots demonstrated that ardipusilloside I caused Mc3 cell death through the induction of apoptosis by downregulation of Bcl-2 protein levels and upregulation of Bax and caspase-3 protein levels.ConclusionsOur results revealed that ardipusilloside I could be a new active substance for mucoepidermoid carcinoma treatment. We demonstrated that the potential mechanism of inhibition might be through the induction of apoptosis by regulation of Bcl-2 family protein levels. This suggests a further rationale for the development of ardipusilloside I as an anti-cancer agent.
Highlights
We showed that baicalin exhibited anticancer activity in mucoepidermoid carcinoma cell line Mc3 by suppressing cell cycle progression and inducing cell apoptosis [5]
Ardipusilloside I inhibited the proliferation of MEC1 cells and Mc3 cells but did not affect the viability of primary cultured parotid acinar cells The effect of ardipusilloside I on the viability of Mc3 cell, another mucoepidermoid carcinoma cell line MEC1 cells and primary cultured parotid acinar cells were assessed by the MTT assay
The results showed that low dose of ardipusilloside I did not affect the viability of primary cultured parotid acinar cells, and 12.5 μg/ml of ardipusilloside I for 48 h showed slight cytotoxicity (Figure 2A)
Summary
DC., family Myrsinaceae, is a traditional Chinese medicine named Jiu Jie Long with a variety of pharmacological functions including anti-cancer activities. We purified a natural triterpenoid saponin, ardipusilloside I, from Ardisia pusilla, and show that it exhibits inhibitory activities in human mucoepidermoid carcinoma Mc3 cells. We investigated the underlying mechanisms of proliferation inhibition that ardipusilloside I exerts on Mc3 cells. To explore new active substances or Ardipusilloside I (3-O-[α-L-rhamnopyranosyl-(1 → 2)-βD-glucopyranosyl-(1 → 3)-(β-D-glucopyranosyl-(1 → 2))-αL-arabinopyranosyl]-cyclamiretin A) (Figure 1) is a natural triterpenoid saponin isolated from Ardisia pusilla A. Laboratory studies on animals and cell lines have shown that ardipusilloside I inhibits cell proliferation [6], induces cell apoptosis [8,9], and inhibits liver cancer survival, invasion, and metastasis both in vitro and in vivo [10]. Little is known about the potential proliferation inhibition mechanism of ardipusilloside I in Mc3 cells
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