Total flavonoids of Radix Tetrastigma suppress inflammation-related hepatocellular carcinoma cell metastasis

Abstract

This study aimed to investigate the effects of the total flavonoids of Radix Tetrastigma (RTF) on inflammation-related hepatocellular carcinoma (HCC) development. Extracted RTF was diluted to different concentrations for subsequent experiments. HCC cells were cotreated with lipopolysaccharide (LPS) and RTF to investigate the effects of RTF on LPS-stimulated HCC cells. A CCK-8 kit was used to measure cell proliferation. Apoptosis was detected with a flow cytometer. Cell migration and invasion were quantified by wound healing and Transwell assays, respectively. The expression of TLR4 and COX-2 and activation of the NF-κB pathway were determined by Western blotting. Treatment with LPS significantly enhanced cell proliferation and decreased the apoptosis rate, while cell migration and invasion were notably upregulated. RTF suppressed the proliferation and invasion induced by LPS stimulation and promoted HCC cell apoptosis. The protein levels of Bax and cleaved caspase-3 were decreased and that of Bcl-2 was increased by LPS in HCC cells, which could be rescued by RTF. RTF significantly inhibited the LPS-induced expression of the proinflammatory mediators IL-6 and IL-8 in HCC cells. Mechanistically, with RTF treatment, the upregulated expression of TLR4 and COX-2 induced by LPS was obviously downregulated. Furthermore, the phosphorylation of NF-κB/p65 was significantly decreased in LPS-stimulated cells after supplementation with RTF. Our study suggests that RTF exerts a significant inhibitory effect on the LPS-induced enhancement of the malignant behaviors of HCC cells via inactivation of TLR4/NF-κB signaling. RTF may be a promising chemotherapeutic agent to limit HCC development and inflammation-mediated metastasis.

The Role of TLR4 in the Immunotherapy of Hepatocellular Carcinoma: Can We Teach an Old Dog New Tricks?

“…In detail, in vitro evidence from HepG2 and SMMC7721 cells has indicated that the activation of mitogenactivated protein kinase kinase 4 (MKK4)/JNK signaling [62] potentiates metastasis, while the combination of in vitro and in vivo evidence from HepG2, Hep3B, and SMMC7721 cell lines implicated the downregulation of NF-κB, resulting in a decrease in TNFα and IL-6 expression with the inhibition of metastasis [61]. The latter was also validated by Liu et al, who further introduced the flavonoids of Radix Tetrastigma, which share NF-κB inhibiting properties, as promising chemotherapeutic regimens to target metastases [63]. A second pro-metastatic mechanism is the formation of NETs [64,65].…”

“…A growing body of evidence implicates the JAK2/STAT3 signaling axis in the progression and invasion of HCC [60]. In this direction, TLR4 signaling contributes to the metastatic capacity of HCC cells in two ways [61][62][63][64][65]. First, the stimulation of the TLR4 signaling pathway by LPS can enhance their metastatic potential either directly through MKK4/JNK and TLR4/NF-κB signaling [62], or indirectly via the upregulation of IL-6 and TNFα [66].…”

AIM2 upregulation promotes metastatic progression and PD‐L1 expression in lung adenocarcinoma

“…The intracellular inflammatory responses have been highly correlated with the metastatic progression in many types of cancer 32–39 . The activation of NLRP3 inflammasome has also been shown to promote cancer metastasis 11,13,15,16,40,41 .…”

“…30 In this study, AIM2 upregulation is dominant for LUAD derived from The intracellular inflammatory responses have been highly correlated with the metastatic progression in many types of cancer. [32][33][34][35][36][37][38][39] The activation of NLRP3 inflammasome has also been shown to promote cancer metastasis. 11,13,15,16,40,41 Nevertheless, AIM2 downregulation has been correlated with a poorer prognosis, probably due to an increased metastatic potential, in renal cell carcinoma 42 and hepatocellular carcinoma.…”

Chromosome‐level reference genome of Tetrastigma hemsleyanum (Vitaceae) provides insights into genomic evolution and the biosynthesis of phenylpropanoids and flavonoids

“…For example, for stilbene synthase ( STS ) genes, most stilbene‐producing plant species (e.g., Sorghum bicolor , Arachis hypogaea ) possess small families of closely related paralogs, while the genome of grapevine contains a large family of putative STS genes (Parage et al., 2012). It is therefore possible that a unique expansion of the STS gene family contributes to the high abundance and chemical diversity of stilbenes throughout Vitaceae species and varieties (Liu et al., 2021; Suprun et al., 2020; Wang et al., 2016). Our previous phylogeographic studies have shown that T. hemsleyanum consists of two lineages in Southwest (SW) and Central‐South‐East (CSE) China (Wang et al., 2015).…”