摘要
目的探讨三苯氧胺Tamoxifen(TAM)诱导ER(-)小鼠乳腺癌细胞MA782细胞凋亡及其分子作用机制。方法TAM体外作用于MA782细胞。流式细胞仪检测细胞凋亡和细胞周期分布情况。MTT法检测细胞增殖活性。免疫组化法检测MA782细胞TGF-β1蛋白表达量,并用病理图像分析软件对蛋白表达进行半定量分析。RT-PCR法检测MA782细胞TGF-β1mRNA表达情况。结果TAM在体外能明显抑制MA782细胞生长,诱导细胞凋亡,上调TGF-β1mRNA及蛋白表达量,免疫组化半定量分析显示TGF-β1平均灰度值为(59.72±0.02),2μmol/L TAM作用48h后无明显变化,72h后上升到(97.1±0.03),6、10μmol/LTAM作用48h后分别上升为(83.2±0.04)、(96.83±0.02),72别上升到(121.75±0.03)、(139.01±0.05),与对照组相比差异极其显著(P<0.01)。RT-PCR检测结果显示2μmol/L TAM组无明显改变,而6、10μmol/LTAM作用于细胞24、48h后,细胞TGF-β1mRNA水平表现出不同程度的上升,与对照组比较有统计学意义(P<0.05或P<0.01)。结论TAM诱导MA782细胞凋亡,其分子作用机制可能与上调细胞TGF-β1mRNA及蛋白表达有关。
Objective To study if tamoxifen （TAM） can induce growth arrest and apoptosis of ER - negative MA782 mouse breast cancer cell line and to explore the molecular machanisims. Methods MA782 cells were Cultured in RPMI1640 medium with TAM. The proliferative activity of cells was detected by MTT methods and cells apoptosis by flowcytometrey methods. The expression of TGF - β1 protein was detected by immunohistochemical methods, the semi - quantity of protein expression was analyzed by pathological image analysis software, The expression of TGF - β1 mRNA was assessed by RT - PCR methods. Results TAM can induce growth arrest and apoptosis of cells. There was no change of TGF - β1 protein in cells with 2 μmoL/L TAM, but with 6 μmoL/L or 10 μmoL/L TAM, the level of TGF - β1 protein increased from （59.72 ±0.02 ）to （83.2 ±0.04） ,（96.83 ±0.02） after 48h and to （121.75 ±0.03） and （139.01±0.05） for 72h. The difference was significant（P 〈0.01）. RT - PCR results showed that the expression of TGF - β1 mRNA increased in cells exposed to 10 μmoL/L TAM for 48 h. The difference was significant compared with control cells（ P 〈 0.01 ）. Conclusion Up -regulation of TGF - β1 mRNA and protein may be one of the mechanisms through which TAM induced growth inhibition and apoptosis of MA782 cell
出处
《青海医学院学报》
CAS
2005年第4期226-230,共5页
Journal of Qinghai Medical College
作者简介
吴建芳（1972-），女，汉族，青海籍，副教授，硕士
