Abstract:

Objective To investigate the effects of praziquantel (PZQ) on and its mechanisms in the proliferation, migration and apoptosis of hepatocellular carcinoma cells. Methods Hep3B human hepatoma cell lines and Hepa1-6 mouse hepatoma cell lines were cultured in vitro. Hep3B and Hepa1-6 cell lines were selected and divided into normal control group, PZQ treatment group, 5-HT_2B inhibitor group (RS127455 treatment group) and 5-HT_2B inhibitor group + PZQ treatment group (RS127445 + PZQ treatment group). Real-time quantitative fluorescent PCR (qRT-PCR) was performed to detect the relative expression of 5-HT_2B mRNA in Hep3B human hepatoma cell lines and Hepa1-6 mouse hepatoma cell lines, and CCK-8 method was used to detect the proliferation of HCC cells. Cell scratch assay, flow cytometry and western blot were used, respectively to determine the migration ability and apoptosis rate of hepatocellular carcinoma cells as well as the expression of apoptosis-related proteins of Bax and Bcl-2. Results The relative expression levels of 5-HT_2B receptor mRNA from Hep3B and Hepal-6 cell lines in normal control group and PZQ treatment group were 1.02±0.09 and 1.01±0.20, 1.36±0.16 and 1.66±0.16, respectively. The relative mRNA expression levels of 5-HT_2B receptor from Hep3B and Hepal-6 cell lines were increased after PZQ treatment (t=3.22, 5.07, both P<0.05). After 48 h of cell culture, the proliferation rates of the two cell lines in PZQ treatment group were (74.00±4.58) % and (77.00±5.29) %, which were lower than those in normal control group (t=9.88, 7.47, both P<0.01). After 72 h of cell culture, the proliferation rates of the two cell lines in PZQ treatment group were (71.00±6.08) % and (67.33±7.57) %, which were lower than those in normal control group (t=7.87, 6.00, both P<0.05) and RS127445+PZQ treatment group (t=5.48, 3.48, both P<0.05). The cell mobility of PZQ treatment group at 48 h was (52.91±3.15) % and (17.28±1.78) %, which was lower than that of normal control group (t=7.86, 13.46, both P<0.01). The cell mobility of the two cell lines was lower in the PZQ treatment group [(58.79±3.25) % and (22.29±5.87) %] than in the normal control group (t=11.65, 9.57, both P<0.05) and RS127445+PZQ treatment group at 72 h (t=3.13, 6.97, both P<0.05). The apoptosis rate of the two cell stains in PZQ treatment group at 72 h was (16.13±0.66) % and (20.70±2.85) %, which were higher than that of normal control group (t=27.82, 5.65, both P<0.01) and RS127445+PZQ treatment group (t=9.54, 4.10, both P<0.01). The relative protein levels of Bax in PZQ treatment group were 1.70±0.18 and 2.23±0.14, respectively, which were higher than those of normal control group (t=2.83, 7.89, both P<0.05) and RS127445+PZQ treatment group (t=9.40, 5.25, both P<0.05). The relative protein expression levels of Bcl-2 were 0.52±0.17 and 0.53±0.02, respectively, which were lower than those of normal control group (t=3.57, 8.39, both P<0.05) and RS127445+PZQ treatment group (t=12.09, 6.12, both P<0.05). Conclusion PZQ can affect the proliferation, migration and apoptosis of hepatocellular carcinoma cells through 5-HT_2B receptor.

Key words: Hepatoma carcinoma cell, Praziquantel, 5-HT_2B, Proliferation, Migration, Apoptosis