苯丁酸钠对人肝癌HepG2.2.15细胞凋亡及乙型肝炎病毒指标的影响

Effects of sodium phenyl butyrate on the apoptosis,expressions of HBsAg and HBeAg and HBV-DNA content of HepG2. 2. 15 cells

目的探讨苯丁酸钠(SPB)对人肝癌HepG2.2.15细胞凋亡及乙型肝炎病毒指标HBsAg、HBeAg表达和HBV-DNA含量的影响。方法采用四甲基偶氮唑盐比色法(MTT)测定不同浓度(1.0、2.0、4.0、8.0μmol/L)SPB处理24、48h的HepG2.2.15细胞的增殖情况;采用流式细胞术分析(2.0、4.0μmol/L)SPB处理24、48h的HepG2.2.15细胞的凋亡率和细胞周期;采用化学发光法检测8.0μmol/L SPB处理72h的HepG2.2.15细胞的上清液中HBsAg和HBeAg含量,RT-PCR法检测该上清液中HBV-DNA水平。结果 SPB能够以时间和剂量依赖方式升高HepG2.2.15细胞的增殖抑制率(P<0.05),其作用24、48h HepG2.2.15细胞的早期和晚期凋亡率、G0/G1细胞比例均高于对照组,S期细胞比例低于对照组(P<0.05);同一浓度下,SPB处理48h HepG2.2.15细胞凋亡率均高于24h(P<0.05)。8.0μmol/L SPB处理72h,HepG2.2.15细胞的上清液中HBsAg、HBeAg含量及HBV-DNA水平分别为40.22±1.57、69.46±1.75和9.34±0.54,均高于对照组的18.33±0.58、34.92±1.26和5.52±0.45,差异有统计学意义(P<0.05)。结论 SPB对人肝癌HepG2.2.15细胞有诱导分化和促进凋亡的作用,并能刺激乙型肝炎病毒的复制,因此在乙型肝炎病毒指标阳性的肝癌患者中临床上应慎用,必要时与抗乙型肝炎病毒药物联合应用。

Objective To investigate the influence of sodium phenyl butyrate（SPB） on the apoptosis,expressions of HBeAg and HBeAg and HBV-DNA content of HepG2.2.15 cells.Methods The MTT assay was used to determine the cell proliferation at 24,48h with different concentrations（1.0,2.0,4.0,8.0μmol/L） of SPB.The flow cytometry（FCM） was employed to measure the cell cycle and cell apoptosis at 24,48h with 2.0,4.0μmol/L SPB.The expressions of HBsAg and HBeAg and HBV-DNA content in supernatants at 72h with 8.0μmol/L SPB were measured by chemiluminescence detection and RT-PCR.Results Treatment with SPB could increase the proliferation inhibition rate at a time-and dose-dependent manner（P 〈 0.05）.The early and late apoptosis rates and the percentage of cells in G0/G1 phase were higher,but the percentage of cells in S phase were lower in SPB group than control group （P 〈 0.05）.The apoptosis rate was higher at 48h than 24h under the same concentration of SPB （P 〈 0.05）.The expressions of HBsAg,HBeAg and HBV-DNA levels were 40.22 ± 1.57,69.46 ± 1.75 and 9.34 ± 0.54 after 72h treatment with 8.0μ mol/L SPB,higher than 18.33 ± 0.58,34.92 ± 1.26 and 5.52 ± 0.45 （P 〈 0.05） in the control group.Conclusion SPB could induce differentiation and promote the apoptosis of HepG2.2.15 cells.SPB could stimulate HBV replication,so it may not be suitable to treat hepatocellular cancer patients with HBV infection.