Ruanjian Sanjie Formula Affects Metastasis of Lung Cancer by Regulating miR-182-5p/EPAS1/EMT Axis

To investigate the specific mechanism by which Ruanjian Sanjie Formula (RJSJ) inhibits the occurrence and metastasis of lung cancer.

This study developed an in vivo model using H460 cells in nude mice to investigate postoperative lung cancer metastasis. Two groups received either RJSJ (0.2 mL) or saline to assess RJSJ's anti-cancer effects, including disease-free survival duration, recurrence-free metastasis rate, number of pulmonary metastases, as well as weight and coefficient of major organs. An in vitro study was conducted to evaluate the effects of RJSJ at concentrations ranging from 0 to 500 µg/mL on the proliferation, migration, and invasion capabilities of H460 and H1975 cell lines. These effects were assessed using the CCK-8 assay for cell proliferation, the wound healing assay for cell migration, and the Transwell assay for cell invasion. RJSJ's chemical profile was analyzed via UPLC-Q-TOF-MS, and its mechanism was explored through network pharmacology. Specific microRNAs and their target genes were identified and analyzed using bioinformatics, qRT-PCR, and Western blot to examine RJSJ's impact on metastasis-related genes and proteins during epithelial-mesenchymal transition (EMT). Virtual docking and molecular simulations were used to evaluate interactions between RJSJ compounds and microRNAs.

RJSJ could inhibit the proliferation, migration, and invasion of lung cancer cells in vitro (P<0.05 or P<0.01), and it had also been shown to prevent lung cancer recurrence and metastasis in vivo (P<0.05); it contains 47 chemical compounds that act on 191 common targets, including 36 core targets. It reduced cancer cell proliferation and invasion, downregulated miR-182-5p, thereby regulating its target gene EPAS1, and regulated EMT-related markers by decreasing the expression of N-cadherin, MMP2, MMP9, Snail, and Twist, while increasing the expression of E-cadherin. Chlorogenic acid was identified as the main anti-tumor component due to its strong binding affinity to nucleic acids.

RJSJ inhibits lung cancer occurrence and metastasis both in vivo and in vitro. Mechanistically, RJSJ suppresses the EMT process by downregulating miR-182-5p and inhibiting EPAS1 expression, thereby restraining lung cancer malignant progression, with chlorogenic acid identified as a key active component.