Jatrorrhizine inhibits mammary carcinoma cells by targeting TNIK mediated Wnt/β-catenin signalling and epithelial-mesenchymal transition (EMT)
Jatrorrhizine inhibits mammary carcinoma cells by targeting TNIK mediated Wnt/β-catenin signalling and epithelial-mesenchymal transition (EMT)
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DOI：
10.1016/j.phymed.2019.153015
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发表时间：
2019-10-01
期刊：
影响因子：
7.9
通讯作者：
Liang, Zongsuo
中科院分区：
文献类型：
--
作者：
Sun, Yanfang;Gao, Xiaoyan;Liang, Zongsuo
背景资料：Traf 2和Nck相互作用丝氨酸蛋白激酶（TNIK）是一种肿瘤靶蛋白，其高表达与乳腺癌细胞的发生发展密切相关。分子对接结果表明，药根碱，原小檗碱生物碱，具有良好的结合亲和力和相互作用与TNIK。方法：采用CRISPR/Cas9技术敲除TNIK基因，通过免疫荧光和免疫印迹技术定性检测药根碱抑制乳腺癌细胞增殖和转移的作用机制。MTT法检测细胞活力，流式细胞仪检测细胞凋亡，集落形成法检测细胞迁移能力，创伤愈合法检测细胞侵袭能力。4 T1/Luc同种移植瘤模型进一步证实了TNIK的抗肿瘤作用。结果：TNIK的靶向敲除可减弱Wnt/β-catenin信号通路和上皮-间质转化（EMT）的表达，药根碱可增强TNIK的这种作用。药根碱还能明显抑制MDA-MB-231、MCF-7和4 T1细胞的增殖，其IC 5c值分别为11.08 ± 1.19 μ M、17.11 ± 4.54 μ M和22.14 ± 2.87 μ M;药根碱还能诱导线粒体功能障碍和早期凋亡，涉及线粒体凋亡途径。4 T1荷瘤小鼠的实验结果进一步证实了药根碱对乳腺癌细胞的增殖和转移有明显的抑制作用，且无明显毒性。结论：药根碱抑制TNIK对Wnt/beta-catenin信号通路和EMT表达的调控，为乳腺癌的靶向治疗提供了一个新的思路。
Background: Traf2 and Nck interacting serine protein kinase (TNIK) is a tumour target protein which its high expression is closely related to the occurrence and development of mammary carcinoma cells. Molecular docking revealed that jatrorrhizine, a protoberberine alkaloid, exhibits good binding affinity and interaction with TNIK. However, the underlying mechanisms of jatrorrhizine targeting TNIK inhibits the proliferation and metastasis of breast cancer cells remain unclear.Methods: To figure out the mechanisms in vitro and in vivo, the CRISPR/Cas9 technology was used to knockout TNIK gene and detected qualitatively by immunofluorescence and immunoblotting assay. The MIT cell viability assay for cytotoxicity test, the apoptosis were detected by flow cytometry, the migration and invasion were evaluated by colony formation, wound healing assay and cell invasion assay, respectively. Anticancer effects were further corroborated by 4T1/Luc homograft tumour model.Results: The results showed that targeted knockout of TNIK that attenuated Wnt/beta-catenin signalling and epithelial-mesenchymal transition (EMT) expression, the effects were potentiated by the addition of jatrorrhizine. Moreover, jatrorrhizine distinctly inhibited the proliferation of MDA-MB-231, MCF-7 and 4T1 cells with IC 5c , values of 11.08 +/- 1.19 mu M, 17.11 +/- 4.54 mu M and 22.14 +/- 2.87 mu M, induced mitochondrial dysfunction and early apoptosis involving mitochondrial apoptotic pathway. These results were further corroborated by the 4T1 tumour-bearing mice, which showed that jatrorrhizine significantly suppressed the proliferation and metastasis of mammary carcinoma cells without obvious toxicity.Conclusion: These findings provide an overall perspective that jatrorrhizine potentially restrains TNIK regulating Wnt/beta-catenin signalling and EMT expression for mammary cancer targeted therapy.