Chenodeoxycholic Acid Enhances the Effect of Sorafenib in Inhibiting HepG2 Cell Growth Through EGFR/Stat3 Pathway

Abstract

BackgroundHepatocellular carcinoma (HCC) is a highly invasive disease with a high mortality rate. Our previous study found that Chenodeoxycholic acid (CDCA) as an endogenous metabolite can enhance the anti-tumor effect. Sorafenib has limited overall efficacy as a first-line agent in HCC, and combined with CDCA may improve its efficacy.MethodsHepG2 cells and Balb/c nude mice were used respectively for in vitro and in vivo experiments. Flow cytometry, Western blotting, HE and immunohistochemical staining and immunofluorescence were used to study the effects of CDCA combined with sorafenib on HepG2 cell growth and apoptosis-related proteins. Magnetic bead coupling, protein profiling and magnetic bead immunoprecipitation were used to find the targets of CDCA action. The effect of CDCA on EGFR/Stat3 signaling pathway was further verified by knocking down Stat3 and EGFR. Finally, fluorescence confocal, and molecular docking were used to study the binding site of CDCA to EGFR.ResultsIn this study, we found that CDCA enhanced the effect of sorafenib in inhibiting the proliferation, migration and invasion of HepG2 cells. Magnetic bead immunoprecipitation and protein profiling revealed that CDCA may enhance the effect of sorafenib by affecting the EGFR/Stat3 signaling pathway. Further results from in vitro and in vivo gene knockdown experiments, confocal experiments and molecular docking showed that CDCA enhances the efficacy of sorafenib by binding to the extracellular structural domain of EGFR.ConclusionThis study reveals the mechanism that CDCA enhances the inhibitory effect of sorafenib on HepG2 cell growth in vitro and in vivo, providing a potential new combination strategy for the treatment of HCC.

Ramucirumab combination with sorafenib enhances the inhibitory effect of sorafenib on HepG2 cancer cells

“…All single treatments down-regulated the expression of MMP-9 , VEGFR2 and EGFR. This is in agreement with previous studies that reported down regulation in the expression of MMP-9 by sorafenib 64 , and panitumumab 65 ; VEGFR by ramucirumab 26 , 66 , sorafenib 64 , and bevacizumab 67 and EGFR by panitumumab 13 and sorafenib 48 . Combining sorafenib with panitumumab significantly down regulated the expression of each of VEGFR and EGFR in HepG2 cancer treated cells.…”

“…In agreement with previous studies, sorafenib treatments for 24, 48 and 72 h exhibited cytotoxic potential towards HepG2 cancer cells 45 – 48 . Both panitumumab and ramucirumab caused a gradual decrease in HepG2 cancer cells’ viability which was more pronounced at ≥ 500 µg/mL concentrations in the 48 h treatment period and at ≥ 31.25 µg/mL concentrations in the 72 h treatment period.…”

“…It plays a critical role in transcriptional regulation of genes that are involved in tumor cell proliferation, survival, migration and invasion into the extracellular matrix 53 , 70 ; constitutive activation of STAT3 is observed in 72.4% of human HCC 58 . It is also involved in the invasion, metastasis during tumor progression and angiogenesis; has anti-apoptosis and inflammatory response 1 , 17 , 48 . Single treatments with sorafenib, bevacizumab, panitumumab and ramucirumab significantly decreased pSTAT3 protein level in HepG2 cancer cells.…”

Exploring the effectiveness of flavone derivatives for treating liver diseases: Utilizing DFT, molecular docking, and molecular dynamics techniques

“…The backbone of flavones can be modified with hundreds of variations, including hydroxylation, O—C-glycosylation, O-methylation, and acylation. Some glycosylated derivatives of flavones have antifungal effects, while the hydroxylation of C 5 , C 7 , C 3′ , and C 4′ in flavonoids can increase bacterial inhibition by flavonoids [26] , [27] , [28] .…”

“…Furthermore, Carla Sá and her team investigated the effect of Luteolin-7-glucoside (L7G) and found that it promotes liver lipolysis through induction of PPAR-α and CPT-represent-docked decreases HMGCR expression, which contributes to lower total and LDL cholesterol levels. Their study suggested that L7G-rich diets may be useful in controlling both NAFLD and CVD [26 , [28] , [29] , [30] . Jian Lu et al.…”