华支睾吸虫分泌排泄蛋白对肝癌细胞株PLC增殖、凋亡及细胞周期的影响

Effect of Clonorchis sinensis excretory/secretory proteins on the proliferation,apoptosis and cell cycle of hepatocarcinoma PLC cells

目的探讨华支睾吸虫分泌排泄蛋白(CsESP)对肝癌细胞株PLC增殖、细胞周期及细胞凋亡的影响。方法 CCK-8实验检测不同浓度CsESP(10、50、100、200μg/ml)对肝癌细胞PLC增殖能力的改变;使用荧光定量PCR(qRT-PCR)检测华支睾吸虫分泌排泄蛋白(CsESP)对肝癌细胞株PLC内细胞周期的相关指标(CyclinB1、CyclinD1、CyclinE1、CDc2、CDK4、CDK2、Rb),细胞凋亡相关指标(Mcl-1、Bcl-2)mRNA表达水平。结果 CCK-8实验显示CsESP(10μg/ml)能明显促进肝癌细胞株PLC增殖能力,qRT-PCR实验检测出CyclinB1、CyclinD1、CyclinE1、CDc2、CDK4、CDK2、Mcl-1、Bcl-2明显高于CsESP 0μg/ml组,Rb明显低于CsESP 0μg/ml组,差异均有统计学意义(P<0.05)。结论 CsESP可促进肝癌细胞株PLC的增殖,其可能与影响细胞周期和抑制细胞凋亡相关。

Objective To explore the effect of Clonorchis sinensis excretory/secretory proteins on the proliferation,apoptosis and cell cycle of hepatocarcinoma PLC cells. Methods PLC cells were treated with different concentrations（10,50,100,and 200 μg/ml）of Cs ESP. The proliferation of PLC cells was determined by CCK-8 assay. The relative mRNA levels of CyclinB1,CyclinD1,CyclinE1,CDc2,CDK4,CDK2,Rb,Mcl-1 and Bcl-2 were detected by qRT-PCR. Results The Clonorchis sinensis excretory/secretory proteins could significantly promote the proliferation of PLC cells.The mRNA levels of CyclinB1,CyclinD1,CyclinE1,CDc2,CDK4,CDK2,Mcl-1 and Bcl-2 were significantly increased in PLC cells then the level of Rb significantly degraded（P〈0.05）. Conclusion Cs ESP could enhance the proliferation of PLC and affect the apoptosis and cell cycle of PLC cells by increasing expression of Cyclin B1,Cyclin D1,Cyclin E1,CDc2,CDK4,CDK2,Mcl-1and Bcl-2 were significantly increased in PLC cells,while the level of Rb was significantly degraded.