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Updated: Feb 12, 2026

In vitro Cell Migration and Invasion Assays
Published on: June 1, 2014
[Pseudolaric acid B induces G2/M arrest and inhibits invasion and migration in HepG2 hepatoma cells]
1Graduate School, First Affiliated Hospital, Jinzhou Medical University, Jinzhou 121000, China.
Abstract:
Objective To investigate the mechanisms of pseudolaric acid B (PAB) blocks cell cycle and inhibits invasion and migration in human hepatoma HepG2 cells. Methods The proliferation effect of PAB on HepG2 cells was evaluated by MTT assay. The effect of PAB on the cell cycle of HepG2 cells was analyzed by flow cytometry. Immunofluorescence cytochemical staining was applied to observe the effect of PAB on the α-tubulin polymerization and expression in HepG2 cells. TranswellTM chamber invasion assay and wound healing assay were performed to detect the influence of PAB on the migration and invasion ability of HepG2 cells. Western blotting was used to determine the expressions of α-tubulin, E-cadherin and MMP-9 in HepG2 cells after treated with PAB. Results PAB inhibited the proliferation of HepG2 cells in a dose-dependent manner and blocked the cell cycle in G2/M phase. PAB significantly changed the polymerization and decreased the expression of α-tubulin. The capacities of invasion and migration of HepG2 cells treated by PAB were significantly depressed. The protein levels of α-tubulin and MMP-9 decreased while the E-cadherin protein level increased. Conclusion PAB can inhibits the proliferation of HepG2 cells by down-regulating the expression of α-tubulin and influencing its polymerization, arresting HepG2 cells in G2/M phase. Meanwhile, PAB also can inhibit the invasion and migration of HepG2 cells by lowering cytoskeleton α-tubulin and MMP-9, and increasing E-cadherin.
Insights
Pseudolaric acid B (PAB) inhibits hepatoma cell proliferation by arresting the cell cycle and affecting α-tubulin. PAB also suppresses cancer cell invasion and migration by altering key proteins.
Area of Science:
- * Pharmacology
- * Molecular Biology
- * Cancer Research
Background:
- * Hepatocellular carcinoma (HCC) remains a significant global health challenge.
- * Identifying novel therapeutic agents for HCC is crucial.
- * Pseudolaric acid B (PAB) is a compound with potential anti-cancer properties.
Purpose of the Study:
- * To elucidate the mechanisms by which PAB affects HepG2 cell proliferation, cell cycle, invasion, and migration.
- * To investigate the role of α-tubulin, E-cadherin, and MMP-9 in PAB's anti-cancer effects.
Main Methods:
- * MTT assay for proliferation.
- * Flow cytometry for cell cycle analysis.
- * Immunofluorescence for α-tubulin polymerization and expression.
- * Transwell and wound healing assays for invasion and migration.
- * Western blotting for protein expression analysis.
Main Results:
- * PAB inhibited HepG2 cell proliferation and arrested the cell cycle at the G2/M phase.
- * PAB altered α-tubulin polymerization and decreased its expression.
- * PAB significantly reduced HepG2 cell invasion and migration.
- * PAB decreased protein levels of α-tubulin and MMP-9, while increasing E-cadherin.
Conclusions:
- * PAB inhibits HepG2 cell proliferation by down-regulating α-tubulin expression and polymerization, leading to G2/M phase arrest.
- * PAB suppresses invasion and migration by reducing cytoskeleton α-tubulin and MMP-9, and increasing E-cadherin expression.
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