Effects of praziquantel on the malignant biological behavior of hepatocellular carcinoma cells through 5-HT2B receptor
Objective To investigate the effects of praziquantel(PZQ)on and its mechanisms in the proliferation,mi-gration 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-HT2B inhibitor group(RS127455 treatment group)and 5-HT2B inhibitor group+PZQ treatment group(RS127445+PZQ treatment group).Real-time quantitative fluorescent PCR(qRT-PCR)was per-formed to detect the relative expression of 5-HT2B mRNA in Hep3B human hepatoma cell lines and Hepa1-6 mouse hep-atoma cell lines,and CCK-8 method was used to detect the proliferation of HCC cells.Cell scratch assay,flow cytome-try and western blot were used,respectively to determine the migration ability and apoptosis rate of hepatocellular carci-noma cells as well as the expression of apoptosis-related proteins of Bax and Bcl-2.Results The relative expression levels of 5-HT2B 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-HT2B 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 high-er 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 carcino-ma cells through 5-HT2B receptor.
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