黄荆子乙酸乙酯提取物对人绒毛膜癌JEG-3细胞增殖与凋亡的影响及其作用机制

摘要

目的:观察黄荆子乙酸乙酯提取物(purified vitexin compound 2,VB2)对人类绒毛膜癌JEG-3细胞体外增殖及凋亡活性,及对凋亡相关基因mTOR/4E-BP1 mRNA表达的影响.方法:应用四甲基偶氮唑蓝(MTT)法检测不同浓度的VB2对JEG-3细胞的体外增殖活性;Hoechst 33258染色观察细胞形态学变化;流式细胞分析方法定量检测不同浓度药物作用后细胞凋亡率;RT-PCR法检测mTOR/4E-BP1 mRNA水平表达的变化.结果:1)2.5,5.0,10.0,20.0,40.0,80.0,160.0 μmol/L的VB2分别作用JEG-3细胞(24,48,72 h)后,其生长抑制率由(6.34±0.41)％增加至(85.89±0.81)％,且与剂量及作用时间呈明显正相关(P＜0.05).2)0,5.0,10.0,20.0 μmol/L的VB2处理JEG-3细胞48 h后,加药组细胞有明显的凋亡形态特征;其凋亡率由(9.26±1.02)％增至(35.55±1.24)％,并呈明显剂量依赖性(P＜0.05);mTOR及4EBP1 mRNA表达水平随着VB2作用浓度增高逐渐降低,二者呈负相关(P＜0.05).结论:VB2能抑制人绒毛膜癌JEG-3细胞增殖,诱导其凋亡,此作用可能与VB2抑制JEG-3细胞mTOR和4E-BP1mRNA的表达有关.%Objective:To investigate the effect of purified vitexin compound 2 (VB2),a noval lignanoid from the acetoacetate extract of Vitex negundo seed on the proliferation and apoptosis as well as the expression of mTOR and 4E-BP1 mRNA signal pathway in human choriocarcinoma JEG-3 cell lines in vitro.Methods:The inhibitory effect of different concentrations of VB2 on JEG-3 cells was examined by methyl thiazolyl tetrazo1ium (MTT) assay.Flow cytormetry was used to analyze the apoptosis after using different concentrations of VB2,and the expression of mTOR and 4E-BP1 mRNA was determined by RT-PCR.Results:The inhibitory rate of JEG-3 cell growth which was cultured with different concentrations of VB2 (2.5,5.0,10.0,20.0,40.0,80.0,and 160.0 μmol/L) for 24,48,or 72 hours increased from (6.34+0.41)％ to (85.89±0.81)％,and it was positively correlated with the dose and time of culture (P＜0.05).VB2 at 5.0,10.0,or 20.0 μmol/L increased the rate of JEG-3 cell apoptosis in vitro from (9.26±1.02)96 to (35.55±1.24)96 after 48 hour culture,which was in a dose dependent manner (P＜0.05),while 5.0,10.0,or 20.0 μmol/L of VB2 down-regulated the mRNA levels of mTOR and 4E-BP1 after 48 hour culture,which presented a significant negative correlation between VB2 and the mRNA levels of mTOR and 4E-BP1(P＜0.05).Conclusion:VB2 can restrain the proliferation of choriocarcinoma cell JEG-3 and induce its apoptosig This effect may be related to the inhibition of VB2 on the mRNA expression of JEG-3 cell mTOR and 4E-BP1.