Angelicin Inhibits NSCLC Tumor Growth via the Inhibition of Cancer-Associated Fibroblasts

Objective: This study aimed to investigate the effect of angelicin on the NSCLC tumor growth.

Background: Accumulating evidence shows that cancer-associated fibroblasts (CAFs) play an important role in tumor progression and metastasis, making CAFs an increasingly attractive target for therapeutic intervention. Targeted therapies against CAFs have been considered to have the potential to significantly improve cancer treatment outcomes, overcome resistance, and improve immune evasion. Angelicin (Ang), an active ingredient isolated from the Chinese herbal medicine Psoralea corylifolia Linn., has been reported to inhibit tumor progression. Due to its natural origin, angelicin has good clinical safety and low toxicity. Further clinical studies and exploration of its role as a CAF inhibitor in difficult-to-treat tumors like as NSCLC are expected to offer up a new channel for cancer treatment. Furthermore, angelicin's low cost and good biocompatibility make it have important application potential in cancer combination therapy, especially when used in combination with traditional therapies such as chemotherapy and immunotherapy, which may significantly improve treatment outcomes and reduce side effects. However, the mechanism of its anti-tumor effect remains poorly defined. The aim of this study was to investigate whether ANG modulates CAF activity to inhibit NSCLC progression.

Methods: NIH3T3 cells are a mouse fibroblast cell line, and the use of NIH3T3 cells as a model for CAFs is mainly due to their natural fibroblast phenotype, ease of culture, good response to stimuli, and ability to simulate the functions of fibroblasts in the tumor microenvironment. NIH3T3 was treated with TGF-β (4ug/ml) and H2O2 (10μM). A conditioned medium was used to study the effect of Ang on tumor growth, invasion, and migration by regulating CAFs.Ang concentrations were set at 12.5, 25, and 50μM for cell cycle experiments and 0, 20, and 40μM for cell migration and invasion experiments. Subcutaneous tumors were established by mixing LLC and NIH3T3 cells to observe the effect of Ang on tumor progression and microenvironment. Fibroblast activity during Ang intervention was monitored by fluorescence-labeled FAPI-04 and 18F-labeled FAPI-04. The molecular pharmacological mechanism of Ang was investigated by RNA sequencing and network pharmacology.

Results: The result showed that Ang significantly inhibited TGF-β and H2O2-induced NIH3T3 transformation, as evidenced by reduced expression of markers such as FAPα and α-SMA. Ang inhibited proliferation, invasion and migration of LLC cells induced by CAFs-conditioned medium. In vivo Experiments showed that Ang greatly inhibited tumor growth in Lewis's lung cancer caused by CAFs. Molecular pharmacological analysis showed that Ang could modulate CAFs activity through multiple targets. These data indicate that Ang has a great potential to reduce CAF activity, interfere with the tumor microenvironment, and inhibit tumor growth. However, these findings still need to be further validated, especially considering the heterogeneity of CAFs, the differences between in vitro and in vivo models, and potential side effects.

Conclusion: Ang reduced the growth, invasion, and migration of lung cancer by inactive CAFs. This provides a rationale for tumor microenvironment-based treatment of lung cancer and clinical translation of Ang. As a potential anti-cancer drug, Ang has shown significant effects in inhibiting cancer-related fibroblasts (CAFs) and disrupting the tumor microenvironment. However, there are still challenges in translating these findings into clinical treatments, such as heterogeneity of the tumor microenvironment, differences in patient responses, and side effects. Therefore, future research should focus on exploring personalized treatment strategies, evaluating the clinical safety and effectiveness of the drug, and delving deeper into the molecular mechanisms and target sites of Ang.