目的:探讨突变型p27基因(p27mutant type gene;p27mt)对恶性黑素瘤A375细胞凋亡的调节作用。方法:利用重组腺病毒Ad-p27mt转染培养的人恶性黑素瘤A375细胞,用3H-TdR掺入法检测细胞增殖;流式细胞术、DNA片段分析法、TUNEL法检测细胞凋亡...目的:探讨突变型p27基因(p27mutant type gene;p27mt)对恶性黑素瘤A375细胞凋亡的调节作用。方法:利用重组腺病毒Ad-p27mt转染培养的人恶性黑素瘤A375细胞,用3H-TdR掺入法检测细胞增殖;流式细胞术、DNA片段分析法、TUNEL法检测细胞凋亡。结果:重组腺病毒Ad-p27mt在MOI≥50时,可达到100%的转导效率。Ad-p27mt转染恶性黑素瘤细胞A375后,3H-TdR掺入法检测发现细胞增殖抑制;流式细胞术检测在G1期前出现亚二倍体凋亡峰;细胞DNA抽提电泳后发现凋亡特征性梯带。TUNEL法检测凋亡指数分别为48.5±3.6(Ad-p27mt组)及4.2±0.8(空白对照组),差异有显著性(P<0.01)。结论:重组腺病毒介导的p27mt基因转移可诱导恶性黑素瘤A375细胞在Gl期阻滞并导致细胞凋亡。展开更多
Suberoylanilide hydroxamic acid (SAHA), an orally administered inhibitor of histone deacetylases, is currently in phase II clinical trials for cutaneous T cell lymphomas (CTCL), but the mechanism of SAHA action is unk...Suberoylanilide hydroxamic acid (SAHA), an orally administered inhibitor of histone deacetylases, is currently in phase II clinical trials for cutaneous T cell lymphomas (CTCL), but the mechanism of SAHA action is unknown. In this study, we investigated the anti-tumor effects of SAHA in CTCL cell lines and freshly isolated peripheral blood lymphocytes (PBL) from CTCL patients with high percentage of circulating malignant T cells. Three cell lines (MJ, Hut78, and HH) and PBL from 11 patients and three healthy donors were treated with SAHA (1, 2.5, and 5 μ M) for 24 and/or 48 h. Apoptosis was determined by flow cytometry analysis of sub- G1 hypodiploid nuclei and/or annexin V binding populations. Acetylated histones and apoptosis-associated proteins were detected by Western blotting. SAHA at 1- 5 μ M for 24 and 48 h induced apoptosis in a concentration- and time-dependent manner in three cell lines: MJ (0% - 7% and 1% - 32% ), Hut78 (4% - 36% and 5% - 54% ), and HH (4% - 67% and 8% - 81% ). SAHA at 1- 5 μ M for 48 h also induced more apoptosis of patients’ PBL than healthy donors’ (15% - 32% versus6% - 13% , p<0.05). SAHA treatment caused an accumulation of acetylated histones (H2B, H3, and H4), an increase of p21 WAF1 and bax proteins, a decrease of Stat6 and phospho-Stat6 proteins, and activation of caspase-3 in CTCL cells. Our data suggest that selective induction of malignant T cell apoptosis and modulation of acetylated histones, p21WAF1, bax, Stat6, and caspase-3 may underlie the therapeutic action of SAHA in CTCL patients.展开更多
维甲酸是诱导分化剂中重要药物,通过增强抑癌基因表达,影响蛋白激酶,调节信号通路,与维甲酸受体结合,受体再与特异性 DNA 序列结合,进而调节其邻近靶基因的表达,以及通过 JNK(c-jun 氨基末端激酶)的磷酸化,诱导视网膜母细胞瘤细胞凋亡,...维甲酸是诱导分化剂中重要药物,通过增强抑癌基因表达,影响蛋白激酶,调节信号通路,与维甲酸受体结合,受体再与特异性 DNA 序列结合,进而调节其邻近靶基因的表达,以及通过 JNK(c-jun 氨基末端激酶)的磷酸化,诱导视网膜母细胞瘤细胞凋亡,促进其分化成正常成熟的细胞。现就维甲酸诱导分化视网膜母细胞瘤的机制研究作一综述。展开更多
文摘目的:探讨突变型p27基因(p27mutant type gene;p27mt)对恶性黑素瘤A375细胞凋亡的调节作用。方法:利用重组腺病毒Ad-p27mt转染培养的人恶性黑素瘤A375细胞,用3H-TdR掺入法检测细胞增殖;流式细胞术、DNA片段分析法、TUNEL法检测细胞凋亡。结果:重组腺病毒Ad-p27mt在MOI≥50时,可达到100%的转导效率。Ad-p27mt转染恶性黑素瘤细胞A375后,3H-TdR掺入法检测发现细胞增殖抑制;流式细胞术检测在G1期前出现亚二倍体凋亡峰;细胞DNA抽提电泳后发现凋亡特征性梯带。TUNEL法检测凋亡指数分别为48.5±3.6(Ad-p27mt组)及4.2±0.8(空白对照组),差异有显著性(P<0.01)。结论:重组腺病毒介导的p27mt基因转移可诱导恶性黑素瘤A375细胞在Gl期阻滞并导致细胞凋亡。
文摘Suberoylanilide hydroxamic acid (SAHA), an orally administered inhibitor of histone deacetylases, is currently in phase II clinical trials for cutaneous T cell lymphomas (CTCL), but the mechanism of SAHA action is unknown. In this study, we investigated the anti-tumor effects of SAHA in CTCL cell lines and freshly isolated peripheral blood lymphocytes (PBL) from CTCL patients with high percentage of circulating malignant T cells. Three cell lines (MJ, Hut78, and HH) and PBL from 11 patients and three healthy donors were treated with SAHA (1, 2.5, and 5 μ M) for 24 and/or 48 h. Apoptosis was determined by flow cytometry analysis of sub- G1 hypodiploid nuclei and/or annexin V binding populations. Acetylated histones and apoptosis-associated proteins were detected by Western blotting. SAHA at 1- 5 μ M for 24 and 48 h induced apoptosis in a concentration- and time-dependent manner in three cell lines: MJ (0% - 7% and 1% - 32% ), Hut78 (4% - 36% and 5% - 54% ), and HH (4% - 67% and 8% - 81% ). SAHA at 1- 5 μ M for 48 h also induced more apoptosis of patients’ PBL than healthy donors’ (15% - 32% versus6% - 13% , p<0.05). SAHA treatment caused an accumulation of acetylated histones (H2B, H3, and H4), an increase of p21 WAF1 and bax proteins, a decrease of Stat6 and phospho-Stat6 proteins, and activation of caspase-3 in CTCL cells. Our data suggest that selective induction of malignant T cell apoptosis and modulation of acetylated histones, p21WAF1, bax, Stat6, and caspase-3 may underlie the therapeutic action of SAHA in CTCL patients.
文摘维甲酸是诱导分化剂中重要药物,通过增强抑癌基因表达,影响蛋白激酶,调节信号通路,与维甲酸受体结合,受体再与特异性 DNA 序列结合,进而调节其邻近靶基因的表达,以及通过 JNK(c-jun 氨基末端激酶)的磷酸化,诱导视网膜母细胞瘤细胞凋亡,促进其分化成正常成熟的细胞。现就维甲酸诱导分化视网膜母细胞瘤的机制研究作一综述。
