Progesterone receptor modulator CDB-2914 down-regulates proliferative cell nuclear antigen and Bcl-2 protein expression and up-regulates caspase-3 and poly(adenosine 5'-diphosphate-ribose) polymerase expression in cultured human uterine leiomyoma cells.
Progesterone receptor modulator CDB-2914 down-regulates proliferative cell nuclear antigen and Bcl-2 protein expression and up-regulates caspase-3 and poly(adenosine 5'-diphosphate-ribose) polymerase expression in cultured human uterine leiomyoma cells.
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DOI：
10.1210/jc.2004-1569
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发表时间：
2005-02
期刊：
影响因子：
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通讯作者：
Qin-Yang Xu;S. Takekida;N. Ohara;Wei Chen;R. Sitruk-Ware;Elof D. B. Johansson;T. Maruo
中科院分区：
文献类型：
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作者：
Qin-Yang Xu;S. Takekida;N. Ohara;Wei Chen;R. Sitruk-Ware;Elof D. B. Johansson;T. Maruo
本研究旨在探讨孕激素受体调节剂CDB-2914对培养的人子宫肌瘤细胞增殖活性和细胞凋亡的影响。分离的子宫肌瘤细胞在添加10%胎牛血清的无酚红DMEM中传代培养120h，然后在没有或存在分级浓度的CDB-2914(10(-9)、10(-8)、10(-7)和10(-6)M)的情况下降至无血清条件下12、24、48和96h。用四甲基偶氮唑蓝(3-(4，5-dimethylthiazol-2-yl)-2，5-diphenyltetrazodium)比色法测定子宫肌瘤细胞的存活数。免疫细胞化学和免疫印迹法检测增殖细胞核抗原(PCNA)的表达。用末端脱氧核苷酸转移酶介导的2‘-脱氧尿嘧啶核苷5’-三磷酸缺口末端标记法检测细胞凋亡。Western印迹法检测caspase-3、裂解多聚(ADP-核糖)聚合酶(PARP)和Bcl2的表达。与未处理的对照组相比，CDB-2914以剂量依赖的方式减少子宫肌瘤细胞的存活数和细胞内的增殖细胞核抗原阳性细胞数，增加子宫肌瘤细胞的TUNEL阳性细胞数。Western印迹分析显示，CDB-2914处理组与未处理组相比，细胞增殖细胞核抗原和Bcl-2蛋白表达显著降低，caspase-3和PARP裂解蛋白表达增加，且呈剂量依赖关系。提示CDB-2914通过下调增殖细胞核抗原的表达抑制子宫肌瘤细胞的增殖，通过上调caspase-3和PARP的裂解表达，下调Bcl-2蛋白的表达，诱导子宫肌瘤细胞凋亡。
The present study was conducted to evaluate the effects of the progesterone receptor modulator CDB-2914 on proliferative activity and apoptosis in cultured human uterine leiomyoma cells. Isolated leiomyoma cells were subcultured in phenol red-free DMEM supplemented with 10% fetal bovine serum for 120 h and then stepped down to serum-free conditions for 12, 24, 48, and 96 h in the absence or presence of graded concentrations of CDB-2914 (10(-9), 10(-8), 10(-7), and 10(-6) M). The number of viable cultured leiomyoma cells was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazodium bromide assay. Proliferating cell nuclear antigen (PCNA) expression was evaluated by immunocytochemistry and Western blot analysis. Apoptosis was examined by terminal deoxynucleotidyl transferase-mediated 2'-deoxyuridine 5'-triphosphate nick end labeling (TUNEL) assay. Caspase-3, cleaved poly(ADP-ribose) polymerase (PARP), and Bcl-2 expression were assessed by Western blot analysis. Compared with untreated control cultures, treatment with CDB-2914 decreased the number of viable cultured leiomyoma cells and the PCNA-positive rate in those cells and increased the TUNEL-positive rate in cultured leiomyoma cells in a dose-dependent manner. Western blot analysis revealed that treatment with CDB-2914 significantly decreased the expression of PCNA and Bcl-2 protein and increased the expression of cleaved caspase-3 and cleaved PARP in a dose-dependent manner compared with untreated control cultures. These results suggest that CDB-2914 inhibits the proliferation of cultured leiomyoma cells by down-regulating PCNA expression and induces apoptosis by up-regulating cleaved caspase-3 and PARP expression and down-regulating Bcl-2 protein expression in those cells.