二硫代氨基甲酸吡咯烷对苦参碱诱导肝癌细胞凋亡的影响

Inhibition of NF-kappa B activity enhanced apoptosis induced by matrine in hepatocellular carcinoma cells

目的观察二硫代氨基甲酸吡咯烷（PDTC）抑制核因子-κB（NF-κB）活化后对苦参碱诱导肝癌细胞HepG2凋亡的影响。方法MTT法观察苦参碱（分0．8、1．0、1．5、2．0、2．5g／L组）及PDTC联合苦参碱对HepG2细胞增殖的抑制作用。将HepG2细胞随机分为细胞对照组、PDTC组（20μmol／L）、苦参碱组（1．5g／L）和PDTC＋苦参碱联合组，流式细胞仪和末端脱氧核苷酸转移酶介导的脱氧三磷酸尿苷缺口末端标记法检测细胞凋亡；电泳迁移率改变实验检测细胞核内NF-κB的活化水平。结果PDTC增强了苦参碱对细胞增殖的抑制作用（F=183．92，P〈0．01）。苦参碱同时具有诱导HepG2细胞凋亡和NF-κB活化的作用；PDTC能显著增加苦参碱诱导的HepG2细胞凋亡和抑制苦参碱诱导的HepG2的NF-κB活化，细胞凋亡率由6．11％±0．81％增加至12．95％±0．020／0（X^2=9．67，P〈0．05），NF-κB活化的灰度值由38．82±0．17降至32．01±0．69（X^2=10．38，P〈0．05）。结论苦参碱诱导HepG2细胞凋亡的同时激活NF-κB；PDTC可通过抑制NF-κB活化，增强苦参碱诱导HepG2细胞凋亡的作用。

Objective To investigate the relationship between activation of nuclear factor-kappa gene binding （NF-κB） and apoptosis induced by matrine in hepatocellular carcinoma cell line HepG2. Methods HepG2 cells were stimulated by different concentrations of matrine （0.8, 1.0, 1.5, 2.0, 2.5 g/L）. The HepG2 cell survival rates were evaluated by MTT assay. Cultured HepG2 cells were implanted in culture flasks and divided into four groups： a control group, a pyrrolidine dithiocarbamate （PDTC） group （20 μmol/L）, a matrine group （1.5 g/L） and a combination group, PDTC （20 μmol/L） ＋ matrine （1.5 g/L） combination group. Apoptosis induced by matrine was analyzed by flow cytometry （FCM） and TUNEL. The DNA-binding activity of NF-κB was determined by electrophoretic mobility shift assay （EMSA）. Results PDTC enhanced the inhibition of matrine on cell proliferation （F = 183.92, P 〈 0.01）. The apoptosis and activation of NF-κB of HepG2 cells were induced by matrine. PDTC significantly suppressed NF-κB activation induced by matrine in HepG2 cells. PDTC increased the apoptosis induced by matrine of the HepG2 cells from 6.11% ± 0.81% to 12.95% ± 0.02%, x^2 = 9.67, P 〈 0.01. Conclusions Matrine could induce apoptosis, and at the same time induce activation of NF-κB in HepG2 cells. PDTC increases the apoptosis in hepatocellular carcinoma cells and it may be related to suppressing NF-κB activation of HepG2 cells.