摘要
目的观察MTH1抑制剂TH588对人乳腺癌MCF-7及MDA-MB-231细胞增殖的影响,探讨其作用机制。方法MTT法检测不同浓度的TH588(0,8,16,32,64,128μmol/L)对乳腺癌MCF-7及MDA-MB-231细胞增殖的抑制作用,倒置显微镜观察TH588处理乳腺癌MCF-7及MDA-MB-231细胞后细胞形态学变化。集落克隆形成抑制实验观察不同浓度的TH588(0,3.2,6.4,12.8μmol/L)对乳腺癌MCF-7及MDA-MB-231细胞增殖的抑制作用。PI单染流式细胞术检测不同浓度TH588(0,32,64,128μmol/L)对乳腺癌MCF-7及MDA-MB-231细胞死亡的影响,Western blot检测凋亡相关蛋白Bcl-2、Bax的表达。结果在128μmol/L的TH588作用下,MCF-7细胞24,48,72 h的存活率分别为(67.32±4.34)%、(56.81±0.93)%和(30.86±1.46)%,MDA-MB-231细胞24,48,72 h的存活率分别为(63.35±0.49)%、(41.11±0.75)%和(29.15±0.85)%,与对照组(0μmol/L)比较,细胞的存活率明显降低(P<0.05)。在倒置显微镜下可观察到TH588作用于乳腺癌细胞后,细胞数目明显减少,细胞形态发生改变,细胞皱缩,有细胞碎片及颗粒物质形成,细胞边缘不清晰。集落克隆实验结果表明,TH588对乳腺癌MCF-7和MDA-MB-231细胞有明显的增殖抑制作用。PI结果显示,128μmol/L TH588处理MCF-7细胞和MDA-MB-231细胞48 h,细胞死亡率分别为52.8%和55.6%,与对照组相比细胞死亡率明显增高(P<0.05)。Western blot结果显示,TH588可下调Bcl-2蛋白的表达,并上调Bax蛋白的表达。结论 TH588对乳腺癌MCF-7和MDA-MB-231细胞有明显的增殖抑制作用,其机制可能是TH588抑制了MTH1的修复作用,激活Bax和抑制Bcl-2,从而引起乳腺癌细胞的凋亡。
Objective To investigate the effect of MTH1 inhibitor TH588 on the proliferation of human breast cancer cells and explore its possible mechanism. Methods MTT assay was used to detect the viability of breast cancer MCF-7 cells and MDA-MB-231 cells after exposed to different concentrations（0,8,16,32,64,128 μmol/L） of TH588,and the morphological changes of MCF-7 and MDA-MB-231 cells were observed under inverted microscope. The colony formation assay was used to detect the viability of breast cancer MCF-7 cells and MDA-MB-231 cells after exposed to different concentrations（ 0,3. 2,6. 4,12. 8 μmol/L） of TH588. After the breast cancer MCF-7 cells and MDA-MB-231 cells were treated with different concentrations of TH588（0,32,64,128 μmol/L）,the cell death rate was analyzed by flow cytometry with PI staining,and the expression of Bcl-2 and Bax protein was analyzed by Western blot. Results The MCF-7 cells survival rates were（67. 32 ± 4. 34） %,（56. 81 ± 0. 93） % and（30. 86 ± 1. 46） % after exposed to 128 μmol/L TH588 for 24,48,72 h,respectively,and the MDA-MB-231 cells survival rates were（63. 35 ± 0. 49） %,（41. 11 ±0. 75） % and（ 29. 15 ± 0. 85） %,respectively. Compared with control group（ 0 μmol/L）,the survival rates of MCF-7 cells and MDA-MB-231 cells were significantly decreased（ P 〈 0. 05）. Under inverted microscope,the numbers of breast cancer MCF-7 cells and MDA-MB-231 cells were significantly reduced after treated with TH588,cell morphology changed,cells were shrinked,cells debris and particulate matter formation appeared,and the cell edges were not clear. PI staining results showed that the mortalities of MCF-7 cells and MDA-MB-231 cells treated with 128 μmol/L TH588 for 48 h were 52. 8% and 55. 6%,which were significantly higher than those treated with 0 μmol/L TH588（ P 〈 0. 05）. The results of Western blot showed that TH588 down-regulated the expression of anti-apoptotic protein Bcl-2,and up-regulated the expression of proapoptotic protein Bax in MCF-7
出处
《山西医科大学学报》
CAS
2017年第10期1013-1018,共6页
Journal of Shanxi Medical University
基金
国家自然科学基金资助项目(81372899
81603155)
安徽省高等学校省级自然科学研究项目重大项目(KJ2016SD39)
安徽省国际合作交流项目(1503062024)
安徽省高校自然科学研究重点项目(KJ2016A486)
