Sun Shuoshuo, Liu Xiyu, Wei Xiao, Zhang Shaohong, Wang Weimin
Affiliated Hospital of Integrated Traditional Chinese and Western Medicine, Nanjing University of Chinese Medicine, Nanjing, China.
The Affiliated Huaian NO. 1 People's Hospital, Nanjing Medical University, Huaian, China.
Food Nutr Res. 2023 May 19;66. doi: 10.29219/fnr.v66.8981. eCollection 2022.
Liver cancer is associated with a high mortality rate worldwide. Hepatocellular carcinoma (HCC) constitutes a large proportion of primary liver cancers, and most of its alterations currently remain untreatable. Diallyl trisulfide (DATS), the main chemical constituent of allicin, affects tumour development by regulating cell apoptosis. Allicin-induced autophagy could contribute to apoptosis in HepG2 cells. We rigorously examined the autophagy-related mechanism of allicin-induced apoptosis in HepG2 cells. We treated HepG2 cells with DATS to explore the effect of DATS on pro-apoptotic autophagy in HepG2 cell lines and examine its specific molecular mechanism.
HepG2 cells were treated with various concentrations of DATS for 24 and 48 h. Subsequently, cell viability was measured using the cell counting kit-8 (CCK-8) assay and cell clone formation assay. The HepG2 cell apoptosis was measured using Hoechst 33258 staining and western blotting. Autophagy and the AMP-activated protein kinase (AMPK)/NAD-dependent deacetylase sirtuin-1 (SIRT1) signalling pathway were detected using western blotting.
Our results indicated that DATS inhibited HepG2 cell growth. Moreover, the ability of DATS to promote apoptosis in HepG2 cells increased with increasing concentration. We verified the phenomenon of DATS-induced autophagy in HepG2 cells and demonstrated that DATS treatment upregulated the protein expression of LC3-II/I. By measuring the expression of potential autophagy stimulators, we documented that DATS could induce pro-apoptotic autophagy by activating the AMPK/SIRT1 signalling pathway.
DATS induced pro-apoptotic autophagy via the AMPK/SIRT1 signalling pathway in the human HCC HepG2 cell line. Our findings further implicate allicin as a potential therapeutic agent against liver tumours in clinical settings, providing a basis for combining allicin with an autophagy agonist for treating liver cancer.
肝癌在全球范围内具有较高的死亡率。肝细胞癌（HCC）占原发性肝癌的很大比例，目前其大多数病变仍无法治疗。二烯丙基三硫化物（DATS）是大蒜素的主要化学成分，通过调节细胞凋亡影响肿瘤发展。大蒜素诱导的自噬可能有助于HepG2细胞凋亡。我们严格研究了大蒜素诱导HepG2细胞凋亡的自噬相关机制。我们用DATS处理HepG2细胞，以探讨DATS对HepG2细胞系促凋亡自噬的影响，并研究其具体分子机制。
用不同浓度的DATS处理HepG2细胞24小时和48小时。随后，使用细胞计数试剂盒-8（CCK-8）检测法和细胞克隆形成检测法测量细胞活力。使用Hoechst 33258染色和蛋白质印迹法检测HepG2细胞凋亡。使用蛋白质印迹法检测自噬和AMP激活的蛋白激酶（AMPK）/烟酰胺腺嘌呤二核苷酸依赖性脱乙酰酶sirtuin-1（SIRT1）信号通路。
我们的结果表明DATS抑制HepG2细胞生长。此外，DATS促进HepG2细胞凋亡的能力随浓度增加而增强。我们证实了DATS诱导HepG2细胞自噬的现象，并证明DATS处理上调了LC3-II/I的蛋白表达。通过测量潜在自噬刺激物的表达，我们记录到DATS可通过激活AMPK/SIRT1信号通路诱导促凋亡自噬。
DATS通过AMPK/SIRT1信号通路在人肝癌HepG2细胞系中诱导促凋亡自噬。我们的研究结果进一步表明大蒜素在临床环境中作为一种潜在的抗肝肿瘤治疗药物，为将大蒜素与自噬激动剂联合用于治疗肝癌提供了依据。
