The Affiliated Hospital of Qingdao University, Jiangsu Road 16, Qingdao, 266003, China.
Pharmacol Rep. 2020 Oct;72(5):1370-1382. doi: 10.1007/s43440-020-00064-8. Epub 2020 Mar 3.
This research aims to explore the effect of alpinumisoflavone (AIF) as an anti-cancer drug for the treatment of patients with hepatocellular carcinoma (HCC).
Cell counting kit-8 (CCK-8) and colony formation assay were used to evaluate the viability of the cells and their clonogenic ability. Cellular migration and their invasion capabilities were detected using the wound-healing and transwell assay, respectively. The release of lactate dehydrogenase (LDH) was detected using the LDH kit. The expression levels of genes in the cells and tumor tissues were examined by qRT-PCR, western blotting, and immunohistochemical techniques. The cells transfected with mRFP-GFP-LC3 adenoviruses were stained to determine their autophagy status. MCC950 (NLRP3 inflammasome inhibitor) and NLRP3 shRNA were used to block NLRP3-mediated pyroptosis. Chloroquine and Atg 5 siRNA were used to inhibit the autophagy of the cells.
AIF suppressed cell proliferation, migration, and invasion capacity of SMMC 7721 and Huh7 cells. The incorporation of AIF induced the formation of NLRP3 inflammasome assembly, pyroptosis, and autophagy of the cells. However, the anti-proliferative and anti-metastatic effects of AIF on the HCC cells were attenuated by NLRP3 inhibitor and knockdown. Furthermore, Atg 5 knockdown inhibited autophagy and enhanced the rate of AIF-induced pyroptosis of the cells. AIF also suppressed tumor growth and increased the levels of pyroptosis-related genes in tumor tissues, which were consistent with in vitro observations.
AIF inhibited HCC cell growth and metastasis by inducing NLRP3 inflammasome-mediated pyroptosis. Furthermore, AIF-induced autophagy augmented pyroptosis in HCC.
本研究旨在探讨白杨素（AIF）作为一种抗癌药物治疗肝细胞癌（HCC）患者的效果。
使用细胞计数试剂盒-8（CCK-8）和集落形成实验评估细胞活力及其克隆形成能力。通过划痕愈合和 Transwell 实验分别检测细胞迁移和侵袭能力。使用乳酸脱氢酶（LDH）试剂盒检测 LDH 的释放。通过 qRT-PCR、western blot 和免疫组织化学技术检测细胞和肿瘤组织中基因的表达水平。用 mRFP-GFP-LC3 腺病毒转染细胞，以确定其自噬状态。使用 MCC950（NLRP3 炎性体抑制剂）和 NLRP3 shRNA 阻断 NLRP3 介导的细胞焦亡。使用氯喹和 Atg5 siRNA 抑制细胞自噬。
AIF 抑制 SMMC7721 和 Huh7 细胞的增殖、迁移和侵袭能力。AIF 的掺入诱导 NLRP3 炎性体组装、细胞焦亡和自噬。然而，NLRP3 抑制剂和敲低削弱了 AIF 对 HCC 细胞的抗增殖和抗转移作用。此外，Atg5 敲低抑制自噬并增强 AIF 诱导的细胞焦亡率。AIF 还抑制肿瘤生长并增加肿瘤组织中与细胞焦亡相关基因的水平，这与体外观察结果一致。
AIF 通过诱导 NLRP3 炎性体介导的细胞焦亡抑制 HCC 细胞生长和转移。此外，AIF 诱导的自噬增强了 HCC 中的细胞焦亡。
