Progress in Cancer Research: Inhibitory Mechanisms and Effects of Erianin on Pancreatic Cancer

Background and Significance of the Study

Pancreatic cancer is a malignant tumor of the digestive tract, characterized by inconspicuous early symptoms, high malignancy, and limited efficacy of conventional radiotherapy and chemotherapy, leading to a very high mortality rate. Statistics show that the 5-year survival rate for pancreatic cancer globally is less than 10%, and it is expected to become the second leading cause of cancer death by 2030. Due to the limitations of current treatment methods, developing new molecular targeted drugs is of urgent importance. In recent years, the natural compound Erianin has emerged in the treatment of various cancers due to its antitumor activity. This study focuses on the anticancer activity and potential mechanisms of Erianin in pancreatic cancer, aiming to provide new therapeutic strategies for pancreatic cancer patients.

Overview of the Study and Methods

This study, conducted by researchers Ruxue Liu, Minghan Qiu, Xinxin Deng, and others, was published in the journal “Cancer Cell International.” The research comprehensively explores the inhibitory effects and molecular mechanisms of Erianin on pancreatic cancer cells through network pharmacology, in vitro cell experiments, RNA sequencing, bioinformatics analysis, and in vivo experiments.

Experimental Design and Study Process

1. Drug Target Prediction

First, the research team used the SMILES chemical structure of Erianin to acquire its potential targets from the Swiss Target Prediction database, and screened gene targets related to pancreatic cancer from databases such as GeneCards and OMIM. Through protein-protein interaction (PPI) network analysis, a target network of pancreatic cancer and Erianin was constructed, identifying 75 intersection targets of Erianin with pancreatic cancer, primarily involving PI3K/AKT, MAPK, and FOXO signaling pathways.

2. In Vitro Anticancer Activity Evaluation

To study the effects of Erianin on the proliferation, migration, and apoptosis of pancreatic cancer cells, the research team selected the human pancreatic cancer cell lines SW1990 and L3.7 and conducted the following experiments:

• Cell Proliferation Inhibition Assay: Using CCK8, colony formation, and 5-ethynyl-2’-deoxyuridine (EdU) assays, the inhibitory effects of Erianin on cell proliferation were assessed. Results showed that Erianin inhibited the proliferation of pancreatic cancer cells in a concentration-dependent manner.

• Cell Migration Inhibition Assay: Through wound healing and Transwell migration assays, it was found that Erianin treatment significantly inhibited the migration ability of pancreatic cancer cells.

• Cell Cycle Analysis: Flow cytometry indicated that Erianin caused pancreatic cancer cells to arrest in the G2/M phase, and further Western Blot experiments showed upregulation of the cell cycle inhibitor p21 expression.

• Apoptosis Assay: Flow cytometry analysis showed that Erianin significantly increased the apoptosis rate of pancreatic cancer cells by promoting reactive oxygen species (ROS) production and reducing mitochondrial membrane potential. Western Blot experiments further confirmed changes in the expression levels of apoptosis-related proteins Bcl-2, Bax, and Cleaved-Caspase3, indicating that Erianin induced apoptosis via the mitochondrial pathway.

3. RNA Sequencing and Pathway Analysis

RNA sequencing analysis revealed that differentially expressed genes (DEGs) in pancreatic cancer cells treated with Erianin were significantly enriched in biological processes such as focal adhesion, cell adhesion, and ATP hydrolysis. KEGG pathway enrichment analysis indicated that DEGs primarily involved PI3K/AKT, FOXO, and MAPK signaling pathways. Western Blot validation showed reduced phosphorylation levels of AKT while activating phosphorylation of key proteins in the MAPK pathway such as ASK1, JNK, and p38, suggesting that Erianin may inhibit pancreatic cancer progression by inhibiting the AKT/FOXO1 pathway and activating the ASK1/JNK/p38 MAPK signaling pathway.

4. Molecular Docking and Protein Thermal Stability Analysis

Through molecular docking analysis, researchers found that Erianin has good binding affinity with AKT and ASK1 proteins. Thermal shift assays (CETSA) further confirmed that Erianin can directly bind to AKT and ASK1 proteins, enhancing their thermal stability and thus reinforcing Erianin’s effect on target proteins.

5. In Vivo Anticancer Activity Validation

In mouse experiments, the research team constructed a subcutaneous tumor model using SW1990 pancreatic cancer cells, and results showed that Erianin significantly inhibited tumor growth, reducing tumor volume and weight. Moreover, HE and TUNEL staining of tumor tissues showed significant apoptosis, and IHC analysis indicated significantly increased phosphorylation levels of p38 and JNK, while phosphorylation of AKT decreased after Erianin treatment. These results jointly validate the anti-pancreatic cancer effects of Erianin.

Results and Major Findings

1. Inhibition of Pancreatic Cancer Cell Proliferation, Migration, and Invasion by Erianin: The experiments demonstrated that Erianin effectively inhibits the proliferation, migration, and epithelial-mesenchymal transition (EMT) of pancreatic cancer cells in vitro.

2. Induction of Apoptosis and Cell Cycle Arrest by Erianin: Erianin induces apoptosis in pancreatic cancer cells, increases ROS production, and causes cell cycle arrest in the G2/M phase. This effect is achieved by upregulating the expression of p21 protein and pro-apoptotic factors Bax, Cleaved-PARP, and Cleaved-Caspase3.

3. Molecular Mechanism: Inhibition of AKT/FOXO1 Pathway and Activation of ASK1/JNK/p38 MAPK Pathway: Erianin directly binds to AKT and ASK1 proteins, inhibiting the PI3K/AKT signaling pathway and activating the ASK1/JNK/p38 MAPK signaling pathway, thus inhibiting the proliferation and migration of pancreatic cancer cells.

4. In Vivo Antitumor Effects: Mouse experiments confirmed that Erianin significantly inhibits pancreatic cancer tumor growth, reducing tumor volume and weight, further verifying its anticancer potential.

Discussion and Research Value

AKT and MAPK signaling pathways play critical roles in the occurrence and progression of many malignant tumors. Erianin regulates cancer cell growth and apoptosis through dual pathways, highlighting its potential as a candidate drug for pancreatic cancer treatment. The study also emphasizes the importance of natural products in cancer drug development. Natural compounds have structural diversity and high biological activity, making them important sources of anticancer drugs.

While the study clarifies the mechanisms by which Erianin inhibits pancreatic cancer, it also suggests the possibility of bidirectional regulatory effects in different cancer cell types, necessitating further research in other tumor types. Additionally, studies on Erianin’s pharmacokinetics, toxicity, and safety in vivo are needed for it to enter clinical application stages.

Conclusion

This study confirms the significant inhibitory effects of Erianin on pancreatic cancer cells, revealing its anticancer mechanism through inhibition of the AKT/FOXO1 pathway and activation of the ASK1/JNK/p38 MAPK pathway. This research provides a theoretical basis for Erianin as a new drug for pancreatic cancer treatment and offers new ideas for targeted therapy of pancreatic cancer.