Zhao Xuan, Ren Limei, Wang Xiaoru, Han Guangxin, Wang Shuo, Yao Qingguo, Qi Yonghao
Department of Chemical Engineering, Shijiazhuang University, Shijiazhuang, Hebei 050035, P.R. China.
Department of Pathology and Key Laboratory of Kidney of Hebei Medical University, Shijiazhuang, Hebei 050035, P.R. China.
Exp Ther Med. 2022 Feb;23(2):181. doi: 10.3892/etm.2021.11104. Epub 2021 Dec 30.
TNF receptor-associated factor 6 (TRAF6) has been reported to be associated with the development of cancer. Nevertheless, the exact role of TRAF6 in cancer remains unclear. The purpose of the present study was to explore the mechanism of 2-benzoyl-3-hydroxy-4-methyl-9H-xanthen-9-one leading to the inhibition of the activation of AKT and TGF-β-activated kinase 1 (TAK1), and to the apoptosis of MCF-7 cells. Using a computational docking program and examination of AKT and TAK1 level changes, a new small molecule was identified, 2-benzoyl-3-hydroxy-4-methyl-9H-xanthen-9-one, which competitively bound to TRAF6. Next, the effect of this new compound on MCF-7 cells' biological behavior was studied . MTT assays were used to investigate cell viability; flow cytometry and invasion assays were performed to detect early apoptosis and invasion in MCF-7 cells, respectively. Immunoprecipitation, western blotting and caspase-3/9 activity assays were carried out to explore changes in protein expression. Briefly, the present data indicated that 2-benzoyl-3-hydroxy-4-methyl-9H-xanthen-9-one could suppress proliferation, induce early apoptosis and inhibit invasion in MCF-7 cells by suppressing the expression of Bcl-2 and promoting the expression of Bax, caspase-9, and caspase-3. These findings indicated that 2-benzoyl-3-hydroxy-4-methyl-9H-xanthen-9-one could induce apoptosis by inhibiting the activation of AKT and TAK1, and affecting the Bcl-2/Bax-caspase-9-caspase-3 pathway by competitively binding with TRAF6.
肿瘤坏死因子受体相关因子6（TRAF6）已被报道与癌症的发生发展有关。然而，TRAF6在癌症中的确切作用仍不清楚。本研究的目的是探讨2-苯甲酰基-3-羟基-4-甲基-9H-呫吨-9-酮导致AKT和转化生长因子-β激活激酶1（TAK1）激活受到抑制以及MCF-7细胞凋亡的机制。通过使用计算对接程序并检测AKT和TAK1水平变化，鉴定出一种新的小分子2-苯甲酰基-3-羟基-4-甲基-9H-呫吨-9-酮，它能与TRAF6竞争性结合。接下来，研究了这种新化合物对MCF-7细胞生物学行为的影响。采用MTT法检测细胞活力；分别进行流式细胞术和侵袭实验以检测MCF-7细胞的早期凋亡和侵袭情况。进行免疫沉淀、蛋白质印迹和半胱天冬酶-3/9活性测定以探索蛋白质表达的变化。简而言之，目前的数据表明，2-苯甲酰基-3-羟基-4-甲基-9H-呫吨-9-酮可通过抑制Bcl-2表达并促进Bax、半胱天冬酶-9和半胱天冬酶-3的表达来抑制MCF-7细胞的增殖、诱导早期凋亡并抑制侵袭。这些发现表明，2-苯甲酰基-3-羟基-4-甲基-9H-呫吨-9-酮可通过抑制AKT和TAK1的激活，并通过与TRAF6竞争性结合来影响Bcl-2/Bax-半胱天冬酶-9-半胱天冬酶-3途径，从而诱导细胞凋亡。
