Sirt1增强人慢性粒系白血病K562细胞耐药性

基础医学与临床 ›› 2011, Vol. 31 ›› Issue (11): 1223-1228.

Sirt1增强人慢性粒系白血病K562细胞耐药性

毛蓓蓓¹,宋崴¹,吕湘¹,陈厚早²,刘德培¹

1. 中国医学科学院北京协和医学院基础医学研究所
2. 中国医学科学院北京协和医学院基础医学研究所医学分子生物学国家重点实验室

收稿日期:2011-09-22 修回日期:2011-09-26 出版日期:2011-11-05 发布日期:2011-11-02
通讯作者: 刘德培 E-mail:liudp@pumc.edu.cn
基金资助:
“国家重点基础研究发展计划”

Sirt1 enhances drug resistance of CML K562 cells

Received:2011-09-22 Revised:2011-09-26 Online:2011-11-05 Published:2011-11-02

摘要/Abstract

摘要： 目的研究Ⅲ类去乙酰化酶Sirt1对人慢性粒系白血病K562细胞耐药性的影响及其机制。方法分别将酶活性缺失突变型Sirt1（H363Y）和Sirt1 shRNA的表达质粒转染K562细胞后，用G418筛选出稳定表达酶活性缺失突变型Sirt1或 Sirt1 shRNA的K562细胞。用H2O2和 etoposide处理筛选出来的稳定细胞株，Western Blot检测凋亡标志物caspase3的剪切及Bax的表达，同时检测DNA损伤的标志物H2A.X磷酸化；在MCF-7和293A中过表达野生型Sirt1，检测H2O2和 etoposide处理后H2A.X的磷酸化。结果在K562细胞中，酶活性缺失突变型Sirt1（H363Y）的表达和Sirt1的干扰均能促进H2O2和 etoposide诱导的caspase3的剪切及Bax的表达并同时显著抑制H2O2和 etoposide诱导的H2A.X磷酸化；在MCF-7和293A细胞中，野生型Sirt1的过表达能明显增强H2O2和 etoposide诱导的H2A.X的磷酸化。结论 Sirt1能保护K562细胞对抗DNA损伤药物诱导的凋亡，增强DNA损伤修复的信号。

关键词: Sirt1, DNA 损伤, 耐药性, K562细胞

Abstract: Objective to explore the function of Sirt1 in drug resistance of CML（chronic myelogenous leukemia）K562 cells. Methods K562 cells stably expressing dominate negative Sirt1 (H363Y) or Sirt1 shRNA were treated with H2O2 and etoposide. Cleaved caspase3, Bax and γ-H2A.X signals were detected by Western blotting; MCF-7 and 293A cells overexpressing wild type Sirt1 were treated with H2O2 and etoposide. γ-H2A.X was detected by Western blotting. Results Sirt1-H363Y and Sirt1 shRNA expression in K562 cells could enhance the cleavage of caspase3 and the expression of Bax, but inhibit the formation ofγ-H2A.X induced by H2O2 and etoposide. On the contrary, overexpression of wild type Sirt1 in MCF-7 and 293A cells promotes the formation ofγ-H2A.X. Conclusion Sirt1 could enhance the drug resistance and signal for DNA repair in K562 cells.

Key words: Sirt1, DNA damage, drug resistance, K562 cells

毛蓓蓓宋崴吕湘陈厚早刘德培. Sirt1增强人慢性粒系白血病K562细胞耐药性[J]. 基础医学与临床, 2011, 31(11): 1223-1228.

[1]	李兰芳谢哲尹娜罗清雅李贵琴毛成毅易萍. 真核翻译延伸因子eEF1A1在卵巢癌中表达升高及其与临床病理相关性[J]. 基础医学与临床, 2016, 36(6): 846-848.
[2]	张澍余腾骅吴诚义. 雌激素调控ABCG2介导的乳腺癌MCF-7细胞化疗敏感性[J]. 基础医学与临床, 2016, 36(10): 1400-1406.
[3]	石瑶郭倩周也涵万丹叶秀峰. 白藜芦醇通过Sirtuins 1通路促进自噬减轻大鼠脑缺血/再灌注损伤[J]. 基础医学与临床, 2015, 35(4): 496-501.
[4]	代安亚王方冯文莉. 腺病毒介导的AIF及其突变体对K562细胞凋亡的影响[J]. 基础医学与临床, 2015, 35(11): 1453-1459.
[5]	董林范翠青朱宁陈梅红. SLC30A3促进人髓性白血病细胞K562向早期红系分化[J]. 基础医学与临床, 2012, 32(7): 773-777.
[6]	鲁云彪陈厚早张冉郑伟李莉张庆军刘德培. SIRT1上调大鼠血管平滑肌细胞p27Kip1的表达[J]. 基础医学与临床, 2010, 30(7): 714-720.