| Objective:to investigate the anti-tumor effect Of amlodipine on murine hepatic cancer H22 in vitro and in vivo and vitro ,and to analyze its possible mechanism.Method: for the effect of amlodipine on the growth of murine hepatic cancer H22 cells , five concentration gradients of amlodipine (28×10-3,14×10-3,7×10-3,3.5×10-3,1.75×10-3mg/mL) were introduced. At the 12th,24th,36th,48 th h after the cells have exposed the drugs , the cells proliferation was evaluated by MTT assay; while the effect of amlodipine on the growth of murine hepatic cancer H22 in vivo,the KM mice bearing inoculum of murine hepatic cancer H22 were introduced by inoculated the tumor to mice armpit subcutaneously,The mice were divided into four groups with different drugs (amlodipine 3 mg / kg.d Ig. amlodipine 10mg/kg.d Ig. ,5-FU 10 mg/kg.3d Ig.and normal saline 0.1 ml/10g . d Ig. respectivly) and treated for 10 days, then,stripped tumors and measured the weight. The cell apoptosis-related genes bcl-2 and bax protein expression of murine hepatic cancer H22 were observed by immunohistochemistry in tumor tissue sections. The cell cycle of murine hepatic cancer H22 cells were detected by Flow cytometry (FCM) at the 48thh after treated by the drugs(1/2IC50 of amlodipine, IC50 of amlodipine, normal saline, and 0.38μmol·L-1 of 5-FU 0.38μmol·L-1 respectivly).The effect of amlodipine on the expression of Cyclin B1 in murine hepatic cancer H22 cells was detected by immunohistochemistry, and The effect of amlodipine on expression of p53 gene murine hepatic cancer H22 cells was detected by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR).Results: amlodipine exhibed a significant anti-proliferative effect on murine hepatic cancer H22 cells by dose and time dependently,with a 3.4μmol·L-1 of median inhibitory concentration (IC50) .high-dose amlodipine (amlodipine 10mg/kg. d ip.) showed significant anti tumor effect on the mice bearing murine hepatic cancer H22 with the inhibitory rate of 40.5%.amlodipine decreased the expression of the apoptosis regulatory genes bcl-2, increased the expression of bax protein , and bcl-2/bax ratio was been downward obviously.amlodipine could significantly arrest the cells cycle in the G2 phase, and could induce apoptosis in the tumor cells with a dose–dependent manner. And the mechanisim of the effect of amlodipine on the cell cycle and apoptosis were involved in the reduction of cell cycle protein Cyclin B1 expression and increase of p53 gene expression. Conclusion: amlodipine exhibited a significant anti-tumor effect on murine hepatic cancer H22 both in vitro and invivo,and these effects were related to the cell cycle arresting and apoptosis activating by up cell cycle related gene Cyclin B1 and bax protein,p53 gene expression,down bax protein expression .
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