二甲双胍联合柔红霉素对人结肠癌细胞增殖及凋亡的影响  

Effect of Metformin Combined with Daunorubicin on Apoptosis and Proliferation of Conlon Cancer Cells

作　　者：祁冰洁[1] 宛蕾[1] 

机构地区：[1]贵州医科大学基础医学院药理学教研室,贵州贵阳550025

出　　处：《解放军药学学报》2016年第1期18-21,共4页Pharmaceutical Journal of Chinese People's Liberation Army

摘　　要：目的探讨二甲双胍联合柔红霉素对人结肠癌细胞(SW480)增殖和凋亡的影响。方法采用MTT法检测药物对细胞增殖抑制的影响;集落克隆形成法检测药物对细胞集落克隆形成的影响;碘化丙啶(PI)单染法检测细胞凋亡率的变化;AnnexinⅤ/PI双染法检测细胞早期凋亡的变化;蛋白质免疫印迹法检测Bax、Bcl-2蛋白的表达。试验设空白对照组、药物单用组(二甲双胍或柔红霉素)、药物合用组(二甲双胍+柔红霉素)。选择低于半数抑制浓度20 mmol·L-1的二甲双胍和0.2μmol·L-1柔红霉素单用或合用作用于SW480细胞。结果不同浓度的二甲双胍单用、两药合用24、48、72 h后的细胞存活率分别为81.23%、68.88%、57.88%、75.32%、60.98%、50.43%;与单用药组比较有著性差异(P<0.05);两药单用、合用24 h细胞凋亡率分别为24.23%、30.6%、54.83%,与对照组比较有显著性差异(P<0.05);两药合用24 h细胞早期凋亡高于药物单用,与对照组比较有显著性差异(P<0.05);两药单用或合用细胞集落克隆形成数量均减少,与对照组比较有显著性差异(P<0.05);药物合用时促进凋亡蛋白Bax的表达高于药物单用、抑制凋亡蛋白Bcl-2的表达低于药物单用。结论二甲双胍联合柔红霉素能够通过诱导细胞的凋亡,增强对SW480细胞的增殖抑制作用,其作用机制可能与增强Bax的表达,下调Bcl-2有关。Objective To observe and study the impact of metformin combined with daunorubicin on apoptosis and proliferation of colon cancer cells（ SW480）. Methods MTT was used to assay the effect of drugs on cell proliferation and inhibition. Colony cloning was used to assay the effect of drugs on colony formation of cloning. Propidium iodide（ PI）was used to assay the change of apoptotic rate; Western blot was used to measure the expression of Bax and BCL-2. The test included the blank control group,the drug alone group（ 20 mmol·L- 1metformin,0. 2 μmol·L- 1daunorubicin）,and drug combination group（ metformin ＋ daunorubicin）. Results At different concentrations of metformin alone or in combination with daunorubicin,the cell survival rate after 24,48 and 72 hours was 81. 23%,68. 88%,57. 88%,75. 32%,60. 98%,50. 43% respectively. There was significant difference compared with single drug group（ P〈0. 05）. With the two drugs used alone or in combination after 24 hours,the apoptosis rate was 24. 23%,30. 6%,54. 83%,respectively.There was significant difference（ P〈0. 05） compared with the control group. When the two drugs were used alone or in combination,the number of colony-forming cell colonies was reduced. Drug combination promoted the expression of apoptotic protein Bax but inhibited the expression of apoptotic protein Bcl-2. Conclusion Metformin combined with daunorubicin can increase the apoptosis of SW480 and inhibit the proliferation of SW480. This mechanism may be related to downregulation of Bcl-2 and enhancement of Bax protein.

关 键 词：结肠癌 二甲双胍 柔红霉素 凋亡 

分 类 号：R965[医药卫生—药理学]