Background: Hepatocellular carcinoma (HCC), as an aggressive cancer with a high mortality rate, needs high-efficiency and low-toxicity drug therapy. Natural products have great potential as candidate lead compounds for the development of new HCC drugs. Crebanine is an isoquinoline alkaloid derived from Stephania with various potential pharmacological effects such as anti-cancer. However, the molecular mechanism underlying crebanine-induced liver cancer cells apoptosis has not been reported. Here, we investigated the effect of crebanine on HCC and identified a potential mechanism of action.Methods: In this paper, we intend to detect the toxic effects of crebanine on hepatocellular carcinoma HepG2 cells through a series of in vitro experiments, including detecting the effects of crebanine on the proliferation of HepG2 cells using the CCK8 method and plate cloning assay, observing the growth status and morphological changes of crebanine on HepG2 cells by inverted microscopy; and using the Transwell method to determine the the effect of crebanine on the migration and invasion ability of HepG2 cells; using Hoechst 33258 assay to stain cancer cells, thus observing the effect of crebanine on the morphology of HepG2 apoptotic cells, and detecting the apoptotic state and level of HepG2 cells by flow cytometry; using ROS kit and JC-1 assay kit to detect the changes of reactive oxygen species and mitochondrial membrane potential of HepG2 The immunofluorescence assay was taken to verify whether crebanine had an effect on the expression of p-FoxO3a in cancer cells; the Wetern blot assay was also used to examine the effect of crebanine on proteins related to the mitochondrial apoptotic pathway and its effect on the regulation of the relative protein expression of AKT/FoxO3a axis; after this, NAC and AKT inhibitor LY294002 were used to cells were pretreated with NAC and AKT inhibitor LY294002, respectively, in order to further validate the inhibitory effect of crebanine.Results: It was shown that crebanine effectively inhibited the growth and capacity of HepG2 cells migration and invasion in a dose-dependent manner. Furthermore, the effect of crebanine on the morphology of HepG2 cells was observed through microscopy. Meanwhile, crebanine induced apoptosis by causing reactive oxygen species (ROS) burst and mitochondrial membrane potential (MMP) disrupt. We found that crebanine could down-regulate Bcl-2 and up-regulate Bax, cleaved-PARP, cleaved-caspase-3 and cleaved-caspase-9, but these effects were overturned by ROS inhibitor N-acetylcysteine (NAC). Crebanine also down-regulated p-AKT and p-FoxO3a, and the PI3K inhibitor LY294002 significantly enhances this effect. We also found that the expression of AKT/FoxO3a signaling pathway was ROS-dependent. As shown by Western blots, NAC could partially attenuate the inhibitory effect of crebanine on AKT and FoxO3a phosphorylation.Conclusion: Based on our results, our results suggest that crebanine, as a compound with potential anticancer activity, has significant cytotoxic effects on hepatocellular carcinoma,and it likely induces apoptosis via ROS in the mitochondrial pathway and simultaneously affects the biological function of HCC via the ROS-AKT-FoxO3a signaling axis.

Hepatocellular carcinoma (HCC) is the most common cancer of the digestive system. Recently, transmembrane proteins (TMEMs) have been extensively studied in different types of malignant tumors. However, the influence of TMEM206 in hepatocellular carcinoma is unclear. The UALCAN database was used to investigate the expression of TMEM206 mRNA in liver cancer tissues and used the Human Protein Atlas (THPA) to study the expression of TMEM206 in HCC tissues. The expression of TMEM206 was measured in normal liver HL-7702 cells and HepG2, SMMC-7721, and Bel-7402 liver cancer cells. Next, a lentivirus was used to knockdown TMEM206 in HepG2 cells. Furthermore, after verifying knockdown, we studied the effect of TMEM206 downregulation on the malignant behavior of HepG2 and on the PI3K/AKT pathway. TMEM206 was highly expressed in liver cancer cells ( p < 0.001). Downregulation of TMEM206 significantly inhibited the proliferation, migration and invasion of HepG2, significantly promoted the apoptosis of HepG2, and inhibited the expression of P-PI3K and P-AKT. TMEM206 can affect the malignant behavior of HCC. And the PI3K/AKT pathway was affected. This study provides new ideas for the treatment of HCC.

Objectives Gastric cancer (GC) is a major global health concern and is difficult to diagnose in the early stage. Ferroptosis is an iron-dependent, novel form of non-apoptotic cell death. In recent years, inducing the upregulation of ferroptosis-related genes has become a promising therapeutic alternative for cancers, especially those resistant to traditional treatments. However, the relationship between ferroptosis-related genes and GC remains to be further elucidated. Methods In the present study, mRNA expression profiles and corresponding clinical data of patients with GC were retrieved from The Cancer Genome Atlas and used as test data. A multigene signature was constructed using the least absolute shrinkage and selection operator Cox regression model. Data of patients with GC from ‘GSE84426’ in the Gene Expression Omnibus database were used as Training data for validation. Results More than half ferroptosis-related genes were differentially expressed in GC tissues and adjacent normal tissue samples (58.43%) in the test data. Univariate Cox regression analysis showed that 16 differentially expressed genes were related to the prognosis of GC (all p < 0.05). Expression profiles of the 16 DGEs were analysed using LASSO Cox regression, and a prognostic model was established by selecting the 10 best genes for λ. These 10 genes were used to construct a 10-gene signature and stratify patients into two risk groups. Based on the median risk score in the test data, patients with GC were divided into high- and low-risk groups ( p < 0.001). Risk score was an independent predictor for overall survival in multivariate Cox regression analyses ( p < 0.001 and <0.01 in the test and training data, respectively; hazard ratio >1). Receiver operating characteristic curve analysis confirmed the predictive capacity of the 10-gene signature. Functional analysis revealed that tumour-infiltrating lymphocytes, antigen-presenting cell co-stimulation, and cytokine-cytokine receptors were enriched; however, the immune status differed between the two risk groups. Conclusion The novel ferroptosis-related gene signature can be used for GC prognosis. In addition, ferroptosis may provide a novel alternative for the diagnosis and treatment of GC.