Effect and Mechanism of Dicoumarin on the Growth and Proliferation of HBx-Mediated Hepatocellular Carcinoma Cells
WANG Qing, CHENG Shengtao, CHEN Juan*
HCC is one of the most common malignant tumors in the world, and more than 50% of which are associated with HBV (hepatitis B virus) infection. The postoperative recurrence rate of hepatitis B-related hepatocellular carcinoma patients is significantly higher than that of uncombined patients. The viral protein HBx is the main factor to promote the progression of hepatitis B-associated hepatocellular carcinoma. Therefore, it is of great significance to search for small molecule compounds that can effectively inhibit the carcinogenic effect of HBx and slow down the disease progression of hepatitis B-related hepatocellular carcinoma. In this study, the role and mechanism of the small molecule compound dicoumarin in the growth and proliferation of hepatoma cells mediated by HBx were preliminarily investigated. Firstly, the concentration range of dicoumarin was determined by MTT assay; then, the effects of dicoumarin on the growth and proliferation of HBV stable expression cells (HepG2.2.15), HBV infected cells (HepG2-NTCP), HBx overexpression cells (HepG2-HBx), HBV transient expression cells (HepG2 HBV-WT) and hepatoma cells (HepG2) were detected by growth curve, CCK-8 and EdU tests; meanwhile, the colony forming ability of cells was detected by plate colony forming assay. Afterwards, the mRNA and protein levels of β-catenin, c-Myc and Cyclin A2 separately were detected by real-time fluorescent quantitative PCR and Western blot. Finally, the dependence of dicoumarin on HBx for its cancer inhibitory effect was detected. The results showed that dicoumarin inhibited the growth, proliferation and colony formation of HepG2.2.15 cells and HepG2-NTCP cells infected with HBV in a concentration dependent manner; simultaneously, dicoumarin significantly inhibited the growth and proliferation of HBx overexpression cells (HepG2-HBx) and HBV transient expression cells (HepG2 HBV-WT). Further mechanism analysis showed that dicoumarin significantly decreased the levels of β-catenin, c-Myc and Cyclin A2, thereby inhibiting cell growth and proliferation. Finally, it was confirmed that the inhibitory effect of dicoumarin on the growth and proliferation of hepatitis B-related liver cancer depended on HBx. In conclusion, this study found that dicoumarin could effectively inhibit the growth and proliferation of hepatitis B-related hepatocellular carcinoma, which was dependent on HBx.
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