School of Life Science and Technology, China Pharmaceutical University, Nanjing, 210009, China.
State Key Laboratory of Molecular Development Biology, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, S2-316 Building 2, West Beichan Road, Chaoyang District, Beijing, 100101, China.
Invest New Drugs. 2021 Oct;39(5):1222-1231. doi: 10.1007/s10637-021-01102-9. Epub 2021 Mar 31.
Many studies have indicated that histone deacetylase inhibitors (HDACis) have a significant antitumor effect in cancer. Here we report a compound named LT-548-133-1 that not only acts as an HDAC inhibitor but also interferes with microtubule assembly to inhibit MCF-7 cell proliferation and induce apoptosis. Consistent with Chidamide, LT-548-133-1 inhibited HDAC activity and increased histone H3 acetylation. But the difference is that it significantly induced cell cycle G2/M arrest while Chidamide caused G0/G1 arrest in MCF-7 cells. By Western blotting, we found the accumulation of CyclinB1 and phosphorylated histone H3 in LT-548-133-1 treated cells. Immunofluorescence based microtubule-repolymerization experiments and immunofluorescence staining of cell microtubules and nuclei showed that LT-548-133-1inhibited microtubule-repolymerization and induced mitotic abnormalities. The decreased expression of Bcl-2 and the increased expression of Bax, p53, p21, and cleaved-Caspase3 indicated the occurrence of apoptosis. Flow cytometry results also showed an increase in the proportion of apoptotic cells after administration of LT-548-133-1 or Chidamide. Therefore, we demonstrated that LT-548-133-1 could act as an HDAC inhibitor while inhibiting microtubule-repolymerization, causing mitosis to be arrested in G2/M. These two effects ultimately lead to proliferation inhibition and apoptosis of MCF-7 cells.
许多研究表明组蛋白去乙酰化酶抑制剂（HDACi）在癌症中有显著的抗肿瘤作用。在这里，我们报告了一种名为 LT-548-133-1 的化合物，它不仅作为 HDAC 抑制剂，还干扰微管组装，抑制 MCF-7 细胞增殖并诱导细胞凋亡。与 Chidamide 一致，LT-548-133-1 抑制 HDAC 活性并增加组蛋白 H3 乙酰化。但不同的是，它显著诱导细胞周期 G2/M 期阻滞，而 Chidamide 则导致 MCF-7 细胞 G0/G1 期阻滞。通过 Western blot，我们发现 LT-548-133-1 处理的细胞中环蛋白 B1 和磷酸化组蛋白 H3 的积累。基于微管重聚合的免疫荧光实验和细胞微管和核的免疫荧光染色显示，LT-548-133-1 抑制微管重聚合并诱导有丝分裂异常。Bcl-2 的表达下调和 Bax、p53、p21 和 cleaved-Caspase3 的表达增加表明发生了细胞凋亡。流式细胞术结果也表明，用 LT-548-133-1 或 Chidamide 处理后，凋亡细胞的比例增加。因此，我们证明 LT-548-133-1 可以作为 HDAC 抑制剂，同时抑制微管重聚合，导致有丝分裂停滞在 G2/M 期。这两种作用最终导致 MCF-7 细胞的增殖抑制和凋亡。
