摘要
目的探讨吡喹酮(praziquantel,PZQ)对肝癌细胞增殖、迁移和凋亡的影响及其作用机制。方法体外培养Hep3B人肝癌细胞株和Hepa1-6小鼠肝癌细胞株,分为正常对照组、PZQ处理组、5-羟色胺2B(5-HT2B)受体抑制剂组(RS127445组)、5-HT2B受体抑制剂+PZQ处理组(RS127445+PZQ处理组)。采用实时荧光定量PCR(qRTPCR)检测Hep3B人肝癌细胞株和Hepa1-6小鼠肝癌细胞株中5-HT2B受体mRNA相对表达水平,CCK-8法检测细胞增殖情况,划痕实验检测细胞迁移能力,流式细胞术检测细胞凋亡率,western blot法检测Bax、Bcl-2凋亡相关蛋白表达量。结果Hep3B人肝癌细胞株和Hepa1-6小鼠肝癌细胞株5-HT2B受体mRNA相对表达水平正常对照组为1.02±0.09和1.01±0.20,PZQ处理组为1.36±0.16和1.66±0.16,经PZQ处理后5-HT_(2B)受体mRNA相对表达水平均增加(t=3.22、5.07,P均<0.05)。PZQ处理组两种细胞株48 h细胞增殖率为(74.00±4.58)%和(77.00±5.29)%,低于正常对照组(t=9.88、7.47,P均<0.01);72 h细胞增殖率为(71.00±6.08)%和(67.33±7.57)%,低于正常对照组(t=7.87、6.00,P均<0.05)和RS127445+PZQ处理组(t=5.48、3.48,P均<0.05)。PZQ处理组两种细胞株48 h细胞迁移率为(52.91±3.15)%和(17.28±1.78)%,低于正常对照组(t=7.86、13.46,P均<0.01);72 h细胞迁移率为(58.79±3.25)%和(22.29±5.87)%,低于正常对照组(t=11.65、9.57,P均<0.05)和RS127445+PZQ处理组(t=3.13、6.97,P均<0.05)。PZQ处理组两种细胞株72 h细胞凋亡率为(16.13±0.66)%和(20.70±2.85)%,高于正常对照组和RS127445+PZQ处理组(t=27.82、5.65、9.54、4.10,P均<0.01);Bax相对蛋白表达水平分别为1.70±0.18和2.23±0.14,高于正常对照组(t=2.83、7.89,P均<0.05)和RS127445+PZQ处理组(t=9.40、5.25,P均<0.05);Bcl-2相对蛋白表达水平分别为0.52±0.17和0.53±0.02,低于正常对照组(t=3.57、8.39,P均<0.05)和RS127445+PZQ处理组(t=12.09、6.12,P均<0.05)。结论PZQ可通过5-HT2B受体对肝癌细胞的增殖、迁移和凋亡造成影响。
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-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 performed to detect the relative expression of 5-HT2B 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-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-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-HT2B receptor.
作者
戴昱婕
孙捷睿
胡婷婷
刘新建
王勇
DAI Yujie;SUN Jierui;HU Tingting;LIU Xinjian;WANG Yong(Department of Pathogen Biology,School of Basic Medical Sciences,Nanjing Medical University,Nanjing 211166,Jiangsu Province,China)
出处
《热带病与寄生虫学》
CAS
2024年第3期157-163,共7页
Journal of Tropical Diseases and Parasitology
基金
国家自然科学基金项目(82072301)。
关键词
肝癌细胞
吡喹酮
5-HT2B
增殖
迁移
凋亡
Hepatoma carcinoma cell
Praziquantel
5-HT2B
Proliferation
Migration
Apoptosis