Ziyu II inhibits CRC cells growth both in vitro andin vivo
To examine whether Ziyu II exhibits an antitumor effect against CRC, CRC cell lines (HCT116, DLD-1, HT29, SW48, SW620, RKO) and noncancerous colorectal cell line (NCM460) were treated with different dose of Ziyu II. MTT assay showed that Ziyu II markedly inhibited the growth of CRC cells in a dose-dependent manner, whereas the IC50 value in NCM460 cells were much higher than those in CRC cells (Fig.1B). Consistently, the proliferation of CRC cells was significantly decreased upon Ziyu II treatment, as evidenced by reduced colony formation and EdU incorporation (Fig.1C-E). Then we performed LDH release assay and found that Ziyu II damaged the integrity of plasma membrane (Fig.1F). In summary, these results demonstrated that Ziyu II inhibits the growth of CRC cells in vitro .
To further ascertain the antitumor effect of Ziyu II on CRC growthin vivo , we generated a CRC xenograft model by subcutaneously inoculating DLD-1 cells into nude mice. As shown in Fig. 1G and H, Ziyu II treatment markedly decreased the size and the weight of xenografts when compared with the control group. In addition, xenografts treated with Ziyu II grew at a slower rate than those treated with placebo (Fig. 1I). Moreover, most of Ziyu II-treated tumors displayed reduced Ki67 staining (Fig.1J and K). Moreover, we found that Ziyu II treatment had no significant effect on the pathological features of major organs, suggesting that Ziyu II has no obvious toxic or adverse effect on mice (Fig. S1). Taken together, these data suggest that Ziyu II inhibits the growth of CRC cells both in vitro andin vivo .