Related Experiment Video
Updated: May 24, 2025
In Vitro Cultivation Techniques for Modeling Liver Organogenesis, Building Assembloids, and Designing Synthetic Tissues using Human Cell Lines
Published on: April 18, 2025
FVTF inhibits hepatocellular carcinoma stem properties via targeting DNMT1/miR-34a-5p/FoxM1 axis
Xiao-Cheng Cao1,2,3,4, Jinwu Peng1,4, Ye-Bei Qiu5
1Department of Pathology, Xiangya Hospital, Central South University, Changsha, 410008, China.
Background:
Fructus Viticis Total Flavonoids (FVTF) is a novel candidate preparation that possesses anticancer activity. However, the role and mechanism of FVTF-inhibiting human hepatocellular carcinoma (HCC) cell stem properties is unclear.
Methods:
Liquid chromatography (LC) in conjugation with mass spectrometer (MS) was used to identify the compounds of FVTF. Tumorsphere and soft agar colony formation ability, cancer stem marker expression levels, CD133+ cell percentage, and a xenograft model were utilized to investigate the impact of FVTF on HCC cells stemness. PCR array and qRT-PCR were conducted to identify differentially expressed cancer stem-related genes and miRNAs between FVTF-treated and untreated HCC cells, respectively. Pyrosequencing was conducted to assess the DNA methylation level of the miR-34a-5p promoter. A luciferase reporter assay was performed to verify whether FoxM1 serves as a direct target of miR-34a-5p. Additionally, immunohistochemistry of an HCC tissue microarray was carried out to assess the expression levels of DNMT1, FoxM1, and miR-34a-5p.
Results:
A total of 26 compounds, including 10 flavones, in FVTF were identified. FVTF significantly reduced the ability of tumorsphere and soft agar colony formation, the levels of CD44 protein and BMI1, OCT4 and SOX2 mRNAs in HCC cells, and in vivo tumor initiation ability of HCC cells. Mechanistically, FVTF inhibited HCC cell stem properties via targeting DNMT1/miR-34a-5p/FoxM1 axis. Clinically, DNMT1 expression was inversely correlated with miR-34a-5p expression, whereas a positive correlation was noted between DNMT1 and FoxM1 expression levels, and high DNMT1 levels, low miR-34a-5p levels, and high FoxM1 levels were associated with cancer recurrence. Furthermore, a combination of DNMT1, miR-34a-5p and FoxM1 served as an independent prognostic indicator influencing both DFS and OS in patients with HCC.
Conclusions:
FVTF inhibits HCC cell stem properties by targeting DNMT1/miR-34a-5p/FoxM1 axis, which is associated with HCC recurrence and prognosis, and FVTF is a prospective treatment drug for human HCC.
Insights
Fructus Viticis Total Flavonoids (FVTF) inhibits hepatocellular carcinoma (HCC) stemness by targeting the DNMT1/miR-34a-5p/FoxM1 axis. This mechanism is linked to HCC recurrence and prognosis, suggesting FVTF as a potential treatment.
Area of Science:
- Oncology
- Molecular Biology
- Pharmacology
Background:
- Hepatocellular carcinoma (HCC) exhibits stem-like properties contributing to its progression.
- Fructus Viticis Total Flavonoids (FVTF) shows anticancer potential, but its effect on HCC stemness is unknown.
Purpose of the Study:
- To investigate the role and mechanism of FVTF in inhibiting stem properties of human HCC cells.
- To explore the clinical relevance of the identified molecular axis in HCC.
Main Methods:
- LC-MS for FVTF compound identification.
- In vitro (tumorsphere, soft agar) and in vivo (xenograft) assays to assess stemness.
- Gene and miRNA expression analysis (PCR array, qRT-PCR), pyrosequencing, and luciferase assays to elucidate the mechanism.
- Immunohistochemistry on HCC tissue microarrays for clinical correlation.
Main Results:
- FVTF significantly suppressed HCC stemness markers (CD44, BMI1, OCT4, SOX2) and tumor initiation.
- FVTF targets the DNMT1/miR-34a-5p/FoxM1 axis to inhibit HCC stem properties.
- Clinical analysis revealed correlations between DNMT1, miR-34a-5p, and FoxM1, identifying them as prognostic indicators for HCC recurrence, DFS, and OS.
Conclusions:
- FVTF effectively inhibits HCC stemness through the DNMT1/miR-34a-5p/FoxM1 pathway.
- This pathway is crucial for HCC recurrence and prognosis.
- FVTF represents a promising therapeutic candidate for human HCC.
More Related Videos
Related Concept Videos
01:22MicroRNAs
01:24Regulation of Angiogenesis and Blood Supply
01:12Loss of Tumor Suppressor Gene Functions
When the tumor suppressor genes develop mutations or are lost, cells start growing out of control, leading to cancer. However, a single functional copy of the tumor suppressor gene is enough for the cells to maintain their normal functions and cell...
02:23Abnormal Proliferation