Cinobufacini is a well-known Chinese medicine extracted from
Venenum Bufonis, also called
Chan Su. It has been used clinically for various
cancers, including
colon cancer. However, the function of
Cinobufacini on
colon cancer invasion and
metastasis, and its underlying molecular mechanism, is still not clear. In this study, we investigated the function and mechanism of
Cinobufacini on
colon cancer invasion and
metastasis both in vitro and in vivo studies. Human
colon cancer cells were cultured. CCK assay was used to detect the effect of
Cinobufacini on
colon cancer cells proliferation. The invasion and migration abilities were observed by transwell assays, and the expression of invasion and migration related genes MMP2, MMP9, and epithelial-to-mesenchymal transition (EMT) relate genes were observed by Western blot assays. An orthotopic xenograft model in nude mice was established using
colon cancer HCT116 cells, and the function of
Cinobufacini on
colon cancer invasion and
metastasis were observed in vivo. We found
Cinobufacini significantly inhibited
colon cancer cell proliferation in a dose/time-dependent manner; the invasion and migration abilities of
colon cancer were decreased after treated with
Cinobufacini. The
metastasis and EMT related genes MMP9, MMP2,
N-cadherin and Snail were obviously down-regulated, while the expression of
E-cadherin was up-regulated
after treatment with
Cinobufacini. The Wnt/β-
catenin signaling pathway related genes were observed using WB,and results show that the expression of β-
catenin, wnt3a, c-myc,
cyclin D1, and MMP7 were all down-regulated after being treated with
cinobufacini, while the expression of APC was up-regulated. In vivo studies of the volume and weight of orthotopic xenograft
tumors showed significantly shrinkage in the
Cinobufacini group compared to the control group. The enterocoelia and liver
metastasis tumors were significantly decreased, and the expression of MMP9, MMP2, and β-
catenin were also down-regulated, while
E-cadherin was up-regulated in vivo after the treatment with
Cinobufacini. Our data proves that
Cinobufacini can inhibit
colon cancer invasion and
metastasis both in vitro and in vivo; the mechanism is related by suppressing the Wnt/β-
catenin signaling pathway and then inhibiting the EMT of CRC.