Jiang P J, Liu J Y, Yang G C, Li J R, Tian X L, Yang S J, Wei J, Zhang X
Department of Hematology, Affiliated Hospital of North Sichuan Medical College, Nanchong 637000, China.
Chongqing Key Laboratory of Hematology and Microenvironment, Medical Center of Hematology, Second Affiliated Hospital, Army Medical University, Chongqing 400037, China.
Zhonghua Xue Ye Xue Za Zhi. 2025 May 14;46(5):437-444. doi: 10.3760/cma.j.cn121090-20240826-00324.
To investigate the anti-tumor effect of the Histone Deacetylase 6 (HDAC6) inhibitor ACY-738 and its underlying mechanisms in Diffuse Large B-cell Lymphoma (DLBCL) . The expression of HDAC6 in various tumors and DLBCL was analyzed using bioinformatics. DLBCL cells were treated with different concentrations of ACY-738. Cell viability, DNA synthesis, and clone formation were assessed by CCK-8 assay, EdU assay, and soft agar assay, respectively. Intracellular reactive oxygen species (ROS) levels were detected by fluorescence microscopy. Morphological changes in cells were observed using transmission electron microscopy (TEM). Mitochondrial ROS levels and apoptosis were measured by flow cytometry. The expression levels of apoptosis-related and autophagy-related proteins were detected by Western blotting. HDAC6 was highly expressed in DLBCL (<0.05). ACY-738 inhibited the proliferation, DNA synthesis, and colony formation of DLBCL cells in a dose-dependent manner (<0.05). Treatment with ACY-738 increased intracellular and mitochondrial ROS levels in DLBCL cells in a dose-dependent manner (<0.05). TEM revealed that after ACY-738 treatment, mitochondria in cells were swollen and ruptured, mitochondrial cristae were reduced or absent, autolysosomes appeared, and features characteristic of apoptosis were observed. Western blotting showed that after ACY-738 treatment, the expression of the anti-apoptotic protein BCL-2 was downregulated, while the expression of Cleaved-PARP, Cleaved caspase-3, and BAX was upregulated (<0.05). The expression of autophagy-related proteins Atg7, Atg3, LC3B, and P62 was downregulated, and the expression of acetylated P53 protein was upregulated (<0.05) . The HDAC6 inhibitor ACY-738 induces mitochondria-dependent apoptosis and autophagy in DLBCL cells by acetylating P53, thereby inhibiting DLBCL cell proliferation.
研究组蛋白去乙酰化酶6（HDAC6）抑制剂ACY-738在弥漫性大B细胞淋巴瘤（DLBCL）中的抗肿瘤作用及其潜在机制。利用生物信息学分析HDAC6在各种肿瘤及DLBCL中的表达情况。用不同浓度的ACY-738处理DLBCL细胞。分别通过CCK-8法、EdU法和软琼脂法评估细胞活力、DNA合成及克隆形成能力。通过荧光显微镜检测细胞内活性氧（ROS）水平。利用透射电子显微镜（TEM）观察细胞形态变化。通过流式细胞术检测线粒体ROS水平及细胞凋亡情况。通过蛋白质免疫印迹法检测凋亡相关蛋白和自噬相关蛋白的表达水平。HDAC6在DLBCL中高表达（<0.05）。ACY-738以剂量依赖性方式抑制DLBCL细胞的增殖、DNA合成及集落形成（<0.05）。ACY-738处理以剂量依赖性方式增加DLBCL细胞内及线粒体ROS水平（<0.05）。TEM显示，ACY-738处理后，细胞内线粒体肿胀、破裂，线粒体嵴减少或消失，自噬溶酶体出现，并观察到凋亡特征。蛋白质免疫印迹法显示，ACY-738处理后，抗凋亡蛋白BCL-2表达下调，而裂解的PARP、裂解的半胱天冬酶-3和BAX表达上调（<0.05）。自噬相关蛋白Atg7、Atg3、LC3B和P62的表达下调，乙酰化P53蛋白表达上调（<0.05）。HDAC6抑制剂ACY-738通过使P53乙酰化诱导DLBCL细胞发生线粒体依赖性凋亡和自噬，从而抑制DLBCL细胞增殖。
