Fraxetin Suppresses Cell Proliferation and Induces Apoptosis through Mitochondria Dysfunction in Human Hepatocellular Carcinoma Cell Lines Huh7 and Hep3B

Abstract

Fraxetin is a coumarin scaffold compound extracted from Fraxinus rhynchophylla. It has antioxidant, anti-inflammatory, hepatoprotective, and antifibrotic effects. Furthermore, fraxetin has anticancer effects in breast and lung cancer. We aimed to evaluate whether fraxetin has anticancer activity in hepatocellular carcinoma (HCC) cells and its underlying mechanism. We demonstrated the anticancer effects of fraxetin in the HCC cell lines Huh7 and Hep3B. We confirmed that fraxetin inhibited cell proliferation (42% ± 10% Huh7; 52% ± 7% Hep3B) by arresting the cell cycle and inducing apoptosis in both cell lines. Moreover, fraxetin increased reactive oxygen species production (221% ± 55% Huh7; 460% ± 73% Hep3B), depolarized the mitochondrial membranes (ΔΨm) (345% ± 160% Huh7; 462% ± 140% Hep3B), and disrupted calcium homeostasis in both HCC cell lines. Chelating calcium ions with BAPTA-AM restored proliferation in fraxetin-treated Huh7 cells but not in Hep3B cells. Fraxetin did not affect the phosphorylation of extracellular-signal-regulated kinase 1/2, whereas it decreased JNK and phosphoinositide 3-kinase signaling. Furthermore, fraxetin and mitogen-activated protein kinase pharmacological inhibitors had synergistic antiproliferative effects on HCC cells. Although our study was limited to in vitro data that require validation, we suggest that fraxetin is a potential therapeutic agent against HCC progression.

Multi-Omics Analysis Revealed a Significant Alteration of Critical Metabolic Pathways Due to Sorafenib-Resistance in Hep3B Cell Lines

“…Further study confirmed that SOD2 overexpression is a biomarker in predicting poorer prognosis with endometrioid ovarian carcinoma and shows extreme resistance to chemotherapeutics-mediated oxidative stress [ 85 ]. The recent publication also demonstrated the anticancer effect of fraxetin, an extract from Fraxinus rhynchophylla, on Hep3B and Huh7 cells by lowering the SOD2 expression and disrupting antioxidant mechanisms in the mitochondria [ 86 ]. Consequently, we hypothesized that SOD2 overexpression in resistant Hep3B cells could be a potential biomarker for drug resistance, as well as critical in improving the effectiveness of the chemotherapeutic.…”

Fraxetin down-regulates polo-like kinase 4 (PLK4) to inhibit proliferation, migration and invasion of prostate cancer cells through the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) pathway

“…Studies have shown that Fraxetin exerted it anti-cancer effect via suppressing the malignant progression of cancer cells. For example, Fraxetin inhibits the proliferation of human liver cancer cell lines Huh7 and Hep3B as well as induces apoptosis through mitochondrial dysfunction [ 2 ]. Fraxetin also inhibits MCF-7 breast cancer cells proliferation of and induces cell apoptosis [ 3 ].…”

“…Modern pharmacological studies have supported that Fraxetin has various pharmacological properties, such as antibacterial and anti-inflammatory, inhibiting oxidative stress, neuroprotection, tumor suppression, and immunomodulation effects [ 1 ]. Accumulating studies have reported the anti-cancer effects of Fraxetin on a variety of tumors, such as liver cancer [ 2 ], breast cancer [ 3 ], colon adenocarcinoma [ 4 ], pancreatic cancer [ 5 ], and glioma [ 6 ]. These studies illustrated the powerful potential of Fraxetin in anti-cancer, but whether Fraxetin exhibited anti-cancer effect on prostate cancer remain to be clarified.…”

Cytotoxicity and Apoptosis Induction of 6,7-Dehydroroyleanone from Taiwania cryptomerioides Bark Essential Oil in Hepatocellular Carcinoma Cells

“…The apoptotic effect of Hep G2 cells treated with different specimens was analyzed using Cytomics FC500 flow cytometry (Beckman Coulter, Miami, FL, USA). Double immunocytochemistry labeling with FL1 (Annexin V; green fluorescence) and FL3 (propidium iodide; red fluorescence) was used to differentiate four populations (four quadrants in Figure 1), including necrotic cells (Q1, red fluorescent), late apoptotic cells (Q2, both green/red fluorescent), alive/healthy cells (Q3, no fluorescent), and early apoptotic cells (Q4, green fluorescent) [29,30].…”