Anthocyanins from Lycium ruthenicum Murray Inhibit HepG2 Cells Growth, Metastasis and Promote Apoptosis and G2/M Phase Cycle Arrest by Activating the AMPK/mTOR Autophagy Pathway

Abstract

Among the most common malignancies in humans, liver cancer ranks third in terms of mortality in the world. Seeking new anticancer drugs or adjuvant chemotherapy drugs from natural products has attracted the attention of many researchers. Lycium ruthenicum Murray (LR), a health food and traditional Chinese medicine, exerts extensive pharmacological properties, of which anthocyanins are one of the key active components. In this research, we explored the antitumor activity and autophagy regulation mechanism of anthocyanins from Lycium ruthenicum Murray (ALR) in HepG2 cells. Our results found that ALR profoundly reduced the cell viability, clone formation, migration, and invasion and promoted apoptosis and G2/M phase arrest of HepG2 cells in a dose-dependent pattern. Further studies confirmed that ALR treatment significantly increased the number of autophagic vacuoles and autophagosomes, upregulated the expression of Beclin-1, p62, LC3-II/LC3-I, and p-AMPK, and concomitantly downregulated the expression of p-mTOR. When autophagy was inhibited by 3-methyladenine (3-MA), ALR-induced proliferation inhibition, invasion, and migration capabilities, as well as apoptosis rate and G2/M phase arrest, were all reversed, and the activities of key proteins in the AMPK/mTOR pathway were all constrained. In summary, the results presented here indicate that ALR may be effective as a natural antitumor agent by activating AMPK and inhibiting the mTOR autophagy pathway in HepG2 cells.

Anthocyanins from Lycium ruthenicum Murray inhibit HCC tumor angiogenesis and promote autophagic apoptosis through AMPK/PI3K/AKT/mTOR pathway

“…Lycium ruthenicum Murray is an optimal botanical resource for anthocyanins extraction due to its high content of these compounds (Zeng et al 2023). Our previous studies have provided evidence that ALR effectively hinders the growth and spread of HepG2 cells, while also promoting cell death and arresting the G2/M phase cycle (Fan et al 2022). However, as far as we know, there is currently no available information on the impact of ALR on angiogenesis in HCC.…”

“…Our prior research has con rmed that ALR effectively suppresses the proliferation and migration of HepG2 cells (Fan et al 2022), but whether it also affects the function of vascular endothelial cells remains unclear. Therefore, we examined the viability, proliferation, metastatic potency, and angiogenic changes in ALR-treated EA.hy926 cells through the assays of CCK-8, colony formation, cell scratch, invasion and tubule formation.…”

“…Studies have shown that numerous anticancer agents are participated in the mechanisms of autophagic cell death (Miller and Thorburn 2021). Our previous research has also shown that ALR effectively inhibited the proliferation and metastasis of HepG2 cells by inducing autophagic apoptosis (Fan et al 2022). In addition, dysregulation of autophagy has been observed to be closely associated with angiogenesis in different types of cancers (Schaaf et al 2019).…”

Natural products targeting macroautophagy signaling in hepatocellular carcinoma therapy: Recent evidence and perspectives

“…In like manner, Lycium ruthenicum Murray (ALR) can activate AMPK signaling pathway and inhibit mTOR autophagy pathway in HepG2 cells to exert antitumor effects. This effect is mainly reflected by upregulating the number of autophagy vacuoles and autophagosomes; increasing the expression of Beclin‐1, p62, LC3‐II/LC3‐I, and p‐AMPK; and inhibiting the expression of p‐mTOR (Fan et al, 2022). Tanshinone I can also activate AMPKα signaling pathway and participate in the regulation of autophagy signal and its main targets are related to LC3‐II, P62, and MDA‐MB‐231 (Zheng et al, 2020).…”