Our previous study demonstrated that VOSL and its main active ingredients can suppress the growth of estrogen receptor positive breast cancer MCF-7 xenografts3. In this study, we used the estrogen receptor negative breast cancer MDA-MB-231 xenograft model to evaluate further the anti-breast cancer efficiency of VOSL in vivo. The results revealed that VOSL and its main active ingredients also can suppress the growth of MDA-MB-231 xenografts, and VOSL and CD exhibited better anti-breast cancer activity than Cos or Dehy treatment alone (Fig. 7A and B). The inhibitory rates of VOSL, CD, Dehy, and Cos on MDA-MB-231 xenografts are 62.75%, 54.94%, 31.63%,and 27.85%, respectively, after intraperitoneal injections for 30 times. In addition, the expression of several key molecules, such as p53, c-Myc, p-AKT, p-14-3-3, in tumor tissues was determined by immunohistochemistry. The results demonstrated that the expression levels of c-Myc and p-AKT were all reduced and the expression levels of p53 and p-14-3-3 were all elevated in the treatment groups compared with the negative control group. Moreover, the CD or VOSL treated groups showed more obvious expression differences of these molecules than the Cos or Dehy treated groups (Fig. 7C and D). The results were consistent with the in vitro results. Therefore, we concluded that combination treatment of Cos and Dehy inhibits breast cancer through c-Myc/p53 and AKT/14-3-3 pathway.