Cinobufacini Inhibits Colon Cancer Invasion and Metastasis via Suppressing Wnt/β-Catenin Signaling Pathway and EMT
Cinobufacini Inhibits Colon Cancer Invasion and Metastasis via Suppressing Wnt/β-Catenin Signaling Pathway and EMT
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DOI：
10.1142/s0192415x20500354
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发表时间：
2020-01-01
影响因子：
5.7
通讯作者：
Chen, Teng
中科院分区：
文献类型：
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作者：
Wang, Jie;Cai, Han;Chen, Teng
华蟾素是从蟾蜍中提取的著名中药，又名蟾酥。临床上已用于治疗多种癌症，包括结肠癌。然而，华蟾素在结肠癌侵袭和转移中的作用及其分子机制仍不清楚。在本研究中，我们通过体外和体内研究探讨了华蟾素对结肠癌侵袭和转移的功能和机制。培养人结肠癌细胞。采用CCK法检测华蟾素对结肠癌细胞增殖的影响。 Transwell实验观察侵袭和迁移能力，Western blot实验观察侵袭和迁移相关基因MMP2、MMP9和上皮间质转化（EMT）相关基因的表达情况。利用结肠癌HCT116细胞建立裸鼠原位异种移植模型，并在体内观察华蟾素对结肠癌侵袭和转移的作用。我们发现华蟾素以剂量/时间依赖性方式显着抑制结肠癌细胞增殖；华蟾素治疗后结肠癌的侵袭和迁移能力下降。华蟾素治疗后转移及EMT相关基因MMP9、MMP2、N-cadherin、Snail表达明显下调，而E-cadherin表达上调。利用WB对Wnt/β-catenin信号通路相关基因进行观察，结果显示华蟾素处理后β-catenin、wnt3a、c-myc、cyclin D1、MMP7的表达均下调，而APC的表达上调。原位异种移植肿瘤的体积和重量的体内研究显示，与对照组相比，华蟾素组显着缩小。华蟾素治疗后，体内肠腔和肝转移瘤明显减少，MMP9、MMP2和β-catenin的表达也下调，而E-cadherin的表达上调。我们的数据证明华蟾素在体外和体内均能抑制结肠癌的侵袭和转移；其机制与抑制Wnt/β-catenin信号通路进而抑制CRC的EMT有关。
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 obviouslydown-regulated, while the expression of E-cadherin was up-regulated after treatment with Cinobufacini. The Wnt/beta-catenin signaling pathway related genes were observed using WB, and results show that the expression of beta-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 beta-catenin were also down-regulated, while E-cadherin was up-regulated in vivo after the treatment with Cinobufacini. Our data proves that Cino- bufacini can inhibit colon cancer invasion and metastasis both in vitro and in vivo; the mechanism is related by suppressing the Wnt/beta-catenin signaling pathway and then inhibiting the EMT of CRC.