三苯氧胺对乳腺癌细胞生长抑制、凋亡诱导作用及机制研究
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摘要
目的研究三苯氧胺对乳腺癌MCF-7细胞生长及凋亡的影响及其作用机制。方法取对数生长期MCF-7细胞,低、中、高剂量实验组分别给予1,2.5,5μg·mL~(-1)三苯氧胺处理,对照组细胞则以等量生理盐水,分别继续培养24,48,72 h。用噻唑蓝(MTT)法及流式细胞术分别检测乳腺癌MCF-7细胞增殖抑制及凋亡情况;用蛋白免疫印迹法检测各组乳腺癌MCF-7细胞存活素(survivin)蛋白表达情况;光学显微镜下观察对照组及低剂量实验组细胞干预72h后细胞形态学变化。结果对照组和低、中、高剂量实验组干预72 h时乳腺癌MCF-7细胞生长抑制率分别为(22.56±2.96)%,(53.45±3.85)%,(54.21±2.89)%,(55.02±3.02)%;乳腺癌MCF-7细胞凋亡率分别为(4.21±1.38)%,(13.55±1.56)%,(13.86±1.57)%,(14.03±1.61)%。与对照组比较,实验组乳腺癌MCF-7细胞生长抑制率及凋亡率均显著升高(P<0.01),但低、中、高剂量实验组间差异无统计学意义(P>0.05);实验组乳腺癌MCF-7细胞生长抑制率及凋亡率随培养时间的延长呈逐渐升高趋势(P<0.01)。干预24 h后,对照组及低剂量实验组乳腺癌MCF-7细胞survivin蛋白表达量分别为1.09±0.03,0.85±0.02;干预48 h分别为1.11±0.03,0.76±0.02;干预72 h分别为1.12±0.02,0.71±0.01,差异均有统计学意义(均P<0.01)。结论三苯氧胺可显著抑制乳腺癌MCF-7细胞生长,并诱导其凋亡,其作用机制可能与下调乳腺癌MCF-7细胞survivin蛋白表达相关。
Objective To explore the effect of growth inhibition,apoptosis induction of tamoxifen on the breast cancer MCF-7 cells and its mechanism. Methods The logarithmic phase MCF-7 cell in experimental-L/M/H groups were treated with 1,2. 5,5 μg · mL~(-1) tamoxifen,the cells in control group were treated with equivalent normal saline. All groups were treated for 24,48,72 h sequentially. The growth inhibition and apoptosis of breast cancer MCF-7 cell were detected by methylthiazolyldiphenyl-tetrazolium( MTT) method and flow cytometry;the expression of survivin protein apoptosis of breast cancer MCF-7 cell in each group was detected by Western Blot,the changes of cell morphology in control group and experimental-L group at 72 h after managed were observed under optical microscope. Results The growth inhibition rates of control group and experimental-L/M/H groups were( 22. 56 ± 2. 96) %,( 53. 45 ± 3. 85) %,( 54. 21 ± 2. 89) %,( 55. 02 ± 3. 02) %; the apoptosis rate were( 4. 21 ± 1. 38) %,( 13. 55 ± 1. 56) %,( 13. 86 ± 1. 57) %,( 14. 03 ± 1. 61) %. Compared with control group,the growth inhibition rate and apoptosis rate of breast cancer MCF-7 cell in experiment group increased significantly( P < 0. 01),while there was no significant difference among experimental-L/M/H groups( P > 0. 05); the growth inhibition rate and apoptosis rate of apoptosis rate of breast cancer MCF-7 cell in experiment groups showed a trend of increasing gradually with the lengthen of action time( P < 0. 01). The expression of survivin protein in control group and experimental-L group at 24 h after managed were 1. 09 ± 0. 03,0. 85 ± 0. 02; and were 1. 11 ± 0. 03,0. 76 ± 0. 02 at 48 h after managed; and were 1. 12 ± 0. 02,0. 71 ± 0. 01 at 72 h after managed,all with significant difference( all P < 0. 01). Conclusion Tamoxifen can inhibit the growth of breast cancer MCF-7 cell,induce its apoptosis,and the action mechanism maybe related to down-regulation of survivin protein of breast cancer MCF-7 cell.
Objective To explore the effect of growth inhibition,apoptosis induction of tamoxifen on the breast cancer MCF-7 cells and its mechanism. Methods The logarithmic phase MCF-7 cell in experimental-L/M/H groups were treated with 1,2. 5,5 μg · mL~(-1) tamoxifen,the cells in control group were treated with equivalent normal saline. All groups were treated for 24,48,72 h sequentially. The growth inhibition and apoptosis of breast cancer MCF-7 cell were detected by methylthiazolyldiphenyl-tetrazolium( MTT) method and flow cytometry;the expression of survivin protein apoptosis of breast cancer MCF-7 cell in each group was detected by Western Blot,the changes of cell morphology in control group and experimental-L group at 72 h after managed were observed under optical microscope. Results The growth inhibition rates of control group and experimental-L/M/H groups were( 22. 56 ± 2. 96) %,( 53. 45 ± 3. 85) %,( 54. 21 ± 2. 89) %,( 55. 02 ± 3. 02) %; the apoptosis rate were( 4. 21 ± 1. 38) %,( 13. 55 ± 1. 56) %,( 13. 86 ± 1. 57) %,( 14. 03 ± 1. 61) %. Compared with control group,the growth inhibition rate and apoptosis rate of breast cancer MCF-7 cell in experiment group increased significantly( P < 0. 01),while there was no significant difference among experimental-L/M/H groups( P > 0. 05); the growth inhibition rate and apoptosis rate of apoptosis rate of breast cancer MCF-7 cell in experiment groups showed a trend of increasing gradually with the lengthen of action time( P < 0. 01). The expression of survivin protein in control group and experimental-L group at 24 h after managed were 1. 09 ± 0. 03,0. 85 ± 0. 02; and were 1. 11 ± 0. 03,0. 76 ± 0. 02 at 48 h after managed; and were 1. 12 ± 0. 02,0. 71 ± 0. 01 at 72 h after managed,all with significant difference( all P < 0. 01). Conclusion Tamoxifen can inhibit the growth of breast cancer MCF-7 cell,induce its apoptosis,and the action mechanism maybe related to down-regulation of survivin protein of breast cancer MCF-7 cell.
引文
[1]颜劲.三苯氧胺与来曲唑在绝经后乳腺癌新辅助内分泌治疗中的疗效对比[J].中国现代药物应用,2013,7(8):88-90.
[2]JHA K,SHUKLA M,PANDEY M.Survivin expression and targeting in breast cancer[J].Surg Oncol,2012,21(2):125-131.
[3]VAN MEERLOO J,KASPERS GJ,CLOOS J.Cell sensitivity assays:the MTT assay[J/OL].Methods Mol Biol,2011,731:237-245.2011-03-24[2017-12-22].https://link.springer.com/protocol/10.1007%2F978-1-61779-080-5_20.
[4]吴晓娜,蒋红兵.流式细胞术的工作原理及其临床应用[J].中国医疗设备,2011,26(3):91-93.
[5]LIU Z Q,MAHMOOD T,YANG P C.Western blot:technique,theory and troubleshooting[J].North Am J Med Sci,2012,4(9):429-434.
[6]THOMAS S,THUM K T,BICAKU E,et al.Addition of a histone deacetylase inhibitor redirects tamoxifen-treated breast cancer cells into apoptosis,which is opposed by the induction of autophagy[J].Breast Cancer Res Treat,2011,130(2):437-447.
[7]夏想厚,谷俊朝.瘦素对三苯氧胺诱导的乳腺癌MCF-7细胞的凋亡抑制作用及相关机制[J].国际外科学杂志,2014,41(10):673-676.
[8]MORIAI R,TSUJI N,MORIAI M,et al.Survivin plays as a resistant factor against tamoxifen-induced apoptosis in human breast cancer cells[J].Breast Cancer Res Treat,2009,117(2):261-271.
[2]JHA K,SHUKLA M,PANDEY M.Survivin expression and targeting in breast cancer[J].Surg Oncol,2012,21(2):125-131.
[3]VAN MEERLOO J,KASPERS GJ,CLOOS J.Cell sensitivity assays:the MTT assay[J/OL].Methods Mol Biol,2011,731:237-245.2011-03-24[2017-12-22].https://link.springer.com/protocol/10.1007%2F978-1-61779-080-5_20.
[4]吴晓娜,蒋红兵.流式细胞术的工作原理及其临床应用[J].中国医疗设备,2011,26(3):91-93.
[5]LIU Z Q,MAHMOOD T,YANG P C.Western blot:technique,theory and troubleshooting[J].North Am J Med Sci,2012,4(9):429-434.
[6]THOMAS S,THUM K T,BICAKU E,et al.Addition of a histone deacetylase inhibitor redirects tamoxifen-treated breast cancer cells into apoptosis,which is opposed by the induction of autophagy[J].Breast Cancer Res Treat,2011,130(2):437-447.
[7]夏想厚,谷俊朝.瘦素对三苯氧胺诱导的乳腺癌MCF-7细胞的凋亡抑制作用及相关机制[J].国际外科学杂志,2014,41(10):673-676.
[8]MORIAI R,TSUJI N,MORIAI M,et al.Survivin plays as a resistant factor against tamoxifen-induced apoptosis in human breast cancer cells[J].Breast Cancer Res Treat,2009,117(2):261-271.