^(99)Tc-MDP与帕米膦酸二钠对MDA-MB-231细胞增殖及凋亡作用的对比研究

The comparative study on proliferation and apoptosis of MDA-MB-231 cells by ^(99)Tc-MDP and pamidronate

目的观察(99)~Tc-亚甲基二膦酸盐(MDP)与帕米膦酸二钠对人乳腺癌 MDA-MB-231细胞生长和凋亡的影响及其作用差异。方法不同剂量(99)~Tc-MDP 与帕米膦酸二钠注射液处理细胞,采用细胞计数及四甲基偶氮唑盐(MTT)法检测 MDA-MB-231细胞增殖;流式细胞仪观察细胞凋亡、检测细胞周期及凋亡相关基因 bcl-2与 bax 的表达。结果 50 μmol/L (99)~Tc-MDP 与帕米膦酸二钠处理MDA-MB-231细胞48 h 即可抑制其增殖,细胞存活率分别为45.9%和64.0%,差异有显著性(P<0.05);50 μmol/L (99)~Tc-MDP 与帕米膦酸二钠均可诱导细胞凋亡,用药后大量 MDA-MB-231细胞被阻滞于 G_0/G_1期和 S 期,细胞凋亡率分别为9.59%和5.96%,差异有显著性(P<0.05);50μmol/L(99)~Tc-MDP 即可使 MDA-MB-231细胞凋亡相关基因 bcl-2表达明显减弱,200 μmol/L 帕米膦酸二钠可使 bcl-2表达明显减弱,二者对 bax 表达无影响。结论 (99)~Tc-MDP 及帕米膦酸二钠均可抑制 MDA-MB-231细胞增殖及诱导 MDA-MB-231细胞凋亡,并调控 bcl-2的表达;(99)~Tc-MDP 作用强于帕米膦酸二钠。

Objective To observe the influence and difference of cell growth and apoptosis effect of ^99Tc-methylene diphosphonic acid （MDP） and pamidronate to human breast cancer cell line MDA-MB-231. Methods Human breast cancer cell line MDA-MB-231 cells were treated with different concentrations of ^99Tc-MDP and pamidronate. The proliferation of cells was studied by cell counting and methyl thiazolyl tetrazolium （MTr） assay. The rate of apoptosis cell cycle and the expression of bcl-2 and bax were observed by flow cytometric assay. Results Concentration of 50 μmol/L ^99Tc-MDP and pamidronate could both inhibit the proliferation of MDA-MB-231 cells （ the 48-hour＇s survival rate was 45.9% and 64. 0% ）, and their difference was significant（ P 〈 0.05 ）. Concentration of 50μmol/L ^99Tc-MDP and pamidronate could both induce apoptosis of MDA-MB-231 cells. The apoptosis rates were 9.59% and 5.96%, respectively, and their difference was significant （ P 〈 0.05 ）. Most of cells were blocked in the G0/G1 phase and S phase. Concentration of 200 μmol/L ^99Tc-MDP and 200 μmol/L pamidronate could both induce apoptosis obviously with expression of bcl-2 gene being down regulated. But the expression of bax gene was no marked regulation. Conclusions ^99Tc-MDP and pamidronate can both induce apoptosis and inhibit proliferation of MDA-MB-231 cells in vitro, and also down regulate the expression of bcl-2 gene. The effect of ^99Tc-MDP is stronger than that of pamidronate.