The anti-proliferation effect of Aikete injection on hepatocellular carcinoma in vitro and in vivo

Context: Aikete injection is composed of acetylshikonin and beta,beta-dimethylacrylshikonin, which have been reported to have anti-tumor effects on a wide range of cancer cell lines. However, little is known about the effects of the combination of the two components on cancer cells. Objective: To investigate the anti-proliferation activity of Aikete injection on human hepatocellular carcinoma (HCC) cells and its mechanism. Materials and methods: 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide assay and growth curve assay were used to determine the inhibitory effect of Aikete injection on the proliferation of SMMC-7721 cells. Giemsa staining, Hoechst 33258 staining and flow cytometry were used to assess cell apoptosis. Expression of Bcl-2 and Bax was analyzed by reverse transcription-polymerase chain reaction and flow cytometry. H22 bearing mice were also used to determine the anti-tumor effect of Aikete injection in vivo. Results: Aikete injection inhibited the proliferation of SMMC-7721 cells in both a dose- and time-dependent manner in vitro. The characteristics of apoptosis were observed in Aikete injection groups by Hoechst 33258 and Giemsa staining. In addition, Aikete injection induced cell cycle arrest at G2/M phase and downregulated the Bcl-2 expression and the ratio of Bcl-2/Bax in SMMC-7721 cells. The experiment in vivo showed that Aikete injection significantly inhibited the growth of H22 carcinoma, with an inhibitory rate of 34.37--57.99%. Discussion and conclusion: The results demonstrated that Aikete injection suppressed the growth of HCC cells in vitro and in vivo by inducing cell apoptosis.