Bai-He-Gu-Jin-Tang formula suppresses lung cancer via AKT/GSK3β/β-catenin and induces autophagy via the AMPK/mTORC1/ULK1 signaling pathway

Abstract

Aims: Bai-He-Gu-Jin-Tang (BHGJT) is a classic Chinese formula used to treat lung cancer, while the underlying molecular mechanism remains obscure. The aim of the study was to investigate the molecular mechanism of BHGJT on lung cancer and demonstrate the potential for synergistic treatment combining BHGJT with conventional therapy. Methods: Cell viability assay, colony formation assay and EdU assay were used to determine the in vitro effects of BHGJT, and a subcutaneous xenograft model was used to evaluate the in vivo effect. Cell cycle analysis, apoptosis rate analysis, immunohistochemical and immunofluorescent staining, Western blot assays and network pharmacology-based analysis were used to explore the underlying mechanisms. Results: We found that BHGJT inhibited cell proliferation via a dose-dependent pathway and obviously hindered tumor growth in vivo in lung cancer. Cell cycle arrest and apoptosis were pronouncedly induced by BHGJT via dysregulation of the cell cycle regulators CDK4 and Cyclin D1 and dysregulation of apoptosis-associated proteins, such as cleaved caspase 3/9 and the BCL-2 family. Based on a network pharmacology-based analysis and experimental evidence, we demonstrated that the AKT/GSK3β/ β-catenin signaling pathways were responsible for BHGJT-induced apoptosis in lung cancer cells. Additionally, autophagy was induced by BHGJT via the AMPK/mTORC1/ULK1 signaling pathway, and blocking autophagy with either chloroquine or a ULK1 inhibitor increased the killing efficiency of BHGJT in lung cancer cells. Conclusion:Our findings indicate that the BHGJT formula efficiently inhibits lung cancer growth and represents a potential complementary and alternative treatment for lung cancer.

The mechanism of Bai He Gu Jin Tang against non-small cell lung cancer revealed by network pharmacology and molecular docking

“…[36] The top GO-biological process term of BHGJT's treatment in NSCLC cells was negative regulation of the apoptotic process, while BHGJT has shown an inhibitory effect on the growth of NSCLC cells by inducing cell apoptosis via the AKT/GSK3β/ β-catenin signaling pathways. [6] Interestingly, most of the key compounds in BHGJT such as kaempferol, quercetin, beta-sitosterol and stigmasterol could also trigger the apoptosis of NSCLC. The top 15 KEGG pathways were mostly enriched in the endocrine-related pathways, the inflammation and hypoxia-related pathways as well as other cancer-related pathways.…”

“…[ 4 ] In this context, Bai He Gu Jin Tang (BHGJT) is one of the most commonly applied formulas for the treatment of lung cancer. [ 5 , 6 ] BHGJT is composed of 10 Chinese medicinal herbs: Radix Rehmanniae Praeparata (Shu Di Huang), Radix Rehmanniae (Sheng Di Huang), Radix Angelicae Sinensis (Dang Gui), Radix Paeoniae Alba (Bai Shao), Radix et Rhizoma Glycyrrhizae (Gan Cao), Radix Platycodonis (Jie Geng), Radix Scrophulariae (Xuan Shen), Bulbus Fritillariae Thunbergii (Beimu), Radix Ophiopogonis (Maidong), and Bulbus Lilii (Bai He). BHGJT is a representative formula for cough patients with Yin deficiency syndrome, characterized by cough with little sputum, hemoptysis, lean body, dry mouth, dry stool, a red tongue with a little coating and thin pulse.…”

Research Progress of Baihe Gujin Decoction in the Treatment of Lung Cancer

“…To date, a large number of experiments have confirmed the deep connection of apoptosis with the unrestricted growth and metastasis of tumor cells. One study 24 demonstrated that Baihe Gujin decoction not only induced significant cell cycle arrest through down-regulation of CDK4 and cell cycle protein D1 in the G0/G1 phase, but 350 also promoted mitochondria-dependent apoptosis through the AKT/GSK3β/ β-catenin signaling pathway. After the successful establishment of a Lewis lung cancer nude mouse model, Lv 25 observed that modified Baihe Gujin decoction combined with radiotherapy inhibited the growth of transplanted tumors and promoted the apoptosis of tumor cells in the mice, and that the mechanism may lie in the down-regulation of the expression of hypoxia-inducible factor-1α (HIF-1α), P53, and survivin mRNA and protein.…”

“…As Table 1 shown, we briefly summarized the potential underlying molecule pathways on Baihe Gujin decoction or its active components that exerted inhibitory effects on lung cancer in vivo. 24,[55][56][57][58][59] According to TCM principles, Baihe Gujin decoction is thought to nourish Yin, hydrate the lungs, and alleviate heat. Besides, the components of this decoction are believed to possess properties that promote lung health by addressing imbalances in the body's Yin and Yang.…”

“…Moreover, Baihe Gujin decoction also triggers autophagy through the AMPK/mTORC1/ULK1 pathway, and blocking autophagy enhances Baihe Gujin decoction's efficacy in lung cancer cells. 24 Furthermore, Baihe Gujin decoction enhances immune responses by boosting the anti-tumor effects of T lymphocytes, NK cells, and macrophages. Notably, the anti-inflammatory properties of Baihe Gujin decoction play a crucial role in reducing various cytokines and inflammatory factors, contributing to its effectiveness in combating lung cancers.…”