CA3 Induces Apoptosis of Hepatocellular Carcinoma HepG2 Cells and Related Mechanism

Objective To investigate the effect of CA3(CIL56) on the proliferation and apoptosis of hepatocellular carcinoma HepG2 cells and its mechanism.

Methods HepG2 cells were cultured in vitro and treated with different concentrations of CA3 (0, 0.25, 0.5, 1.0, 2.0, 4.0 mmol/L) for 24 and 48 hours, respectively. Enhanced CCK-8 reagent was used to detect the effect of CA3 on cell proliferation. Flow cytometry was utilized to detect cell apoptosis rate. We detected the changes of intracellular reactive oxygen species (ROS). Western blot was utilized to analyze the expression of YAP1, Bcl-2, Bax and Caspase-3 protein.

Results The enhanced CCK-8 reagent test results showed that after 0.25, 0.5, 1.0, 2.0, 4.0 mmol/L CA3 treatment for 24 and 48 hours, the cell proliferation rates were (80.5±0.3)%, (79.4±0.2)%, (76.2±0.2)%, (76.4±0.1)%, (49.3±0.4)% and (75.3±0.2)%, (64.8±0.3)%, (48.4±0.2)%, (32.2±0.4)%, (31.9±0.2)%, respectively. Flow cytometry results showed that when CA3 concentration was increased to 2 mmol/L, the apoptosis rate was increased to 58.48%. CA3 could significantly increase the content of reactive oxygen species in HepG2 cells, reduce the expression of YAP1 and Bcl-2 protein, and increase the expression of Bax and Caspase-3.

Conclusion CA3 could inhibit the proliferation and induce the apoptosis of HepG2 cells. The mechanism may be related to the increase of intracellular reactive oxygen species production and the regulation of YAP1, Bcl-2, Bax and Caspase-3 protein expression.