氨氯地平对小鼠肝癌H_(22)细胞细胞周期及相关蛋白表达的影响

目的探讨氨氯地平对小鼠肝癌H22细胞细胞周期及相关蛋白表达的影响。方法用不同浓度的氨氯地平作用小鼠肝癌H22细胞,采用MTT法检测细胞的增殖水平,流式细胞术检测细胞周期和凋亡情况,免疫组化及RT-PCR法检测细胞周期蛋白CyclinB1和肿瘤抑制基因p53在蛋白水平和基因水平的表达。结果氨氯地平(1.75×10-3、3.5×10-3、7×10-3、14×10-3、28×10-3mg/ml)作用24、36、48h,均可抑制细胞增殖,作用48h的半数抑制浓度(IC50)为13.4μmol/L;氨氯地平对小鼠肝癌H22细胞细胞周期G2期具有阻滞作用且可使细胞发生凋亡;氨氯地平作用后的小鼠肝癌H22细胞周期蛋白CyclinB1的表达明显降低,p53表达明显增高,差异均有统计学意义(P<0.05)。结论氨氯地平能显著抑制小鼠肝癌H22细胞增殖,并通过下调G2期周期蛋白CyclinB1水平和上调肿瘤抑制基因p53的表达而达到抗肿瘤效果。

Effect of Amlodipine on Cell Cycle of Murine Hepatocarcinoma H22 Cells and Expression of Relevant Protein

Objective To investigate the effect of amlodipine on cell cycle of murine hepatocarcinoma H22 cells and the expression of relevant protein.Methods H22 cells were treated with amlodipine at various concentrations,then determined for proliferation level by MTT method,for cell cycle and apoptosis by flow cytometry,and the expressions of Cyclin B1 and tumor-inhibiting gene p53 at protein and gene levels by immunohistochemical assay and RT-PCR.Results The treatment with amlodipine at various concentrations(1.75 × 10-3,3.5 × 10-3,7 × 10-3,14 × 10-3 and 28 × 10-3 mg/ml)for 24,36 and 48 h inhibited the cell proliferation.The IC50 of amlodipine for treatment for 48 h was 13.4 μmol/L.Amlodipine arrested H22 cells at G2 phase and induced the cell apoptosis.The expression level of Cyclin B1 in the H22 cells after treatment with amlodipine decreased significantly,while that of p53 increased significantly,which showed significant difference with those of blank control(P < 0.01).Conclusions Amlodipine inhibited the proliferation of H22 cells significantly,and showed anti-tumor effect through down-regulating the expression of Cyclin B1 and up-regulating the expression of p53 gene.