CKII抑制剂——肝癌治疗新靶点

目的探讨蛋白激酶（casein kinase 2，cK2）抑制剂影响肝癌细胞株HepG-2凋亡的机制，为其临床治疗肝癌提供可能的实验依据。方法本研究通过MTT法和流式细胞仪检测和观察不同作用时间、不同浓度CK2抑制剂四溴苯三唑（4，5，6，7-tetrabromobenzotriazole，TBB）作用HepG-2肝癌细胞株后对其增殖和凋亡的影响，分别应用荧光定量PCR和Western blot实验分析可能机制。结果TBB在48h浓度为200μmol／L时对肝癌细胞的抑制作用最明显；150~mol／LTBB作用48h后，早期凋亡率明显增高，至（13．2±0．67）％，磷酸化P65在100μmol／LTBB实验组表达下调，而caspase-3、caspase-8、Bcl-2和Bcl—X等在两组间表达未见明显差异。结论CK2抑制剂TBB能够通过参与调控NF—KB信号转导通路抑制肿瘤细胞增殖，促进其凋亡，可能成为未来肝癌治疗的一个新的靶点。

CKII inhibitor TBB inhibits proliferation and induces apoptosis of human hepatic cancer HepG-2 cells in vitro

Objective To investigate the effect of casein kinase 2 （ CK2 ） inhibitor 4,5,6,7- tetrabromobenzotriazole（ TBB ）on cell proliferation and apoptosis of liver cancer cells. Methods Cultured human hepatic cancer HepG-2 cells were treated with different concentrations of TBB. Cell proliferation and apoptosis were examined with MTT assay and flow cytometry, respectively, mRNAs and proteins of Bcl-2, Bcl-xl, Caspase3, Caspase8, PP65 were detected with real-time fluorescence quantitative PCR （RT-PCR）and Western blot, respectively. Results MTT assay showed that TBB markedly inhibited the proliferation of HepG-2 cells at 48 h on the concentration of 200 p, mol/L. 150 μmol/L TBB increased the percentage of early apoptotic cancer cells to （ 13. 2 ±0. 67 ） %. Compared to controls, mRNA and protein expression of phospho-p65 （ PP65 ） was down-regulated in 100 μmol/L TBB treated groups, while there were no significant differences in caspase-3, caspase-8, Bcl-2 and Bcl-x expression between TBB treatment groups and controls. Conclusion TBB inhibits the proliferation and induces the apoptosis of HepG-2 cells through NF-KB pathway, indicating that CK2 may be a novel therapeutic target for liver cancer.