Curcumin reduces expression of Bcl-2, leading to apoptosis in daunorubicin-insensitive CD34+ acute myeloid leukemia cell lines and primary sorted CD34+ acute myeloid leukemia cells.
Curcumin reduces expression of Bcl-2, leading to apoptosis in daunorubicin-insensitive CD34+ acute myeloid leukemia cell lines and primary sorted CD34+ acute myeloid leukemia cells.
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姜黄素降低 Bcl-2 的表达，导致柔红霉素不敏感的 CD34 急性髓系白血病细胞系和原代分选的 CD34 急性髓系白血病细胞凋亡。
DOI：
10.1186/1479-5876-9-71
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发表时间：
2011-05-19
影响因子：
7.4
通讯作者：
Liu Q
中科院分区：
文献类型：
--
作者：
Rao J;Xu DR;Zheng FM;Long ZJ;Huang SS;Wu X;Zhou WH;Huang RW;Liu Q
急性髓系白血病（acute myeloid leukemia，AML）是一种免疫表型异质性的恶性疾病，CD 34阳性与预后不良相关。CD 34 + AML细胞对柔红霉素（DNR）的抗性是CD 34- AML细胞的10-15倍。姜黄素是姜黄的主要成分，在多种癌症中显示出细胞毒性活性;然而，其抗癌活性尚未在DNR不敏感的CD 34 + AML细胞中得到充分研究。因此，本研究的目的是探索姜黄素在DNR不敏感的CD 34 + AML细胞系（KG 1a，Kasumi-1），DNR敏感的U937 AML细胞和原代CD 34 + AML骨髓源性细胞中诱导的细胞毒性。使用CD 34 MicroBead试剂盒从外周血单核细胞或骨髓单核细胞中分离原代人CD 34+细胞。采用MTT法和集落形成实验检测姜黄素的生长抑制作用。碘化丙啶（PI）法检测细胞周期分布;通过Wright-Giemsa、Hoechst 33342和Annexin-V/PI染色分析细胞凋亡。JC-1染色和流式细胞术检测线粒体膜电位（MMP）的变化。逆转录-聚合酶链反应和蛋白质印迹法检测凋亡相关蛋白的表达。Bcl-2的短干扰RNA（siRNA）用于与/不与DNR孵育的CD 34 + KG 1a和Kasumi-1细胞中。姜黄素对柔红霉素不敏感的KG 1a、Kasumi-1和柔红霉素敏感的U937细胞均有增殖抑制作用，并诱导细胞凋亡和G1/S期阻滞。姜黄素诱导的细胞凋亡与Bcl-2 mRNA和蛋白的表达减少，随后的MMP的损失，以及caspase-3的激活，随后的PARP降解有关。姜黄素协同增强DNR不敏感的KG 1a和Kasumi-1细胞中DNR的细胞毒性作用，与Bcl-2表达降低一致。因此，针对Bcl-2的siRNA增加了KG 1a和Kasumi-1细胞对DNR诱导的细胞凋亡的易感性。更重要的是，姜黄素抑制Bcl-2的表达，选择性抑制增殖，并协同增强DNR在原代CD 34 + AML细胞中的细胞毒性，而在正常CD 34+造血祖细胞中显示有限的致死性。姜黄素下调Bcl-2并诱导DNR不敏感的CD 34 + AML细胞系和原代CD 34 + AML细胞凋亡
Acute myeloid leukemia (AML) is an immunophenotypically heterogenous malignant disease, in which CD34 positivity is associated with poor prognosis. CD34+ AML cells are 10-15-fold more resistant to daunorubicin (DNR) than CD34- AML cells. Curcumin is a major component of turmeric that has shown cytotoxic activity in multiple cancers; however, its anti-cancer activity has not been well studied in DNR-insensitive CD34+ AML cells. The aim of this study was to therefore to explore curcumin-induced cytotoxicity in DNR-insensitive CD34+ AML cell lines (KG1a, Kasumi-1), DNR-sensitive U937 AML cells, and primary CD34+ AML bone-marrow-derived cells. Primary human CD34+ cells were isolated from peripheral blood mononuclear cells or bone marrow mononuclear cells using a CD34 MicroBead kit. The growth inhibitory effects of curcumin were evaluated by MTT and colony-formation assays. Cell cycle distribution was examined by propidium iodide (PI) assay. Apoptosis was analyzed by Wright-Giemsa, Hoechst 33342 and Annexin-V/PI staining assays. The change in mitochondrial membrane potential (MMP) was examined by JC-1 staining and flow cytometry. Expression of apoptosis-related proteins was determined by reverse transcription-polymerase chain reaction and Western blotting. Short interfering RNA (siRNA) against Bcl-2 was used in CD34+ KG1a and Kasumi-1 cells incubated with/without DNR. Curcumin inhibited proliferation and induced apoptosis and G1/S arrest in both DNR-insensitive KG1a, Kasumi-1 and DNR-sensitive U937 cells. Curcumin-induced apoptosis was associated with reduced expression of both Bcl-2 mRNA and protein, subsequent loss of MMP, and activation of caspase-3 followed by PARP degradation. Curcumin synergistically enhanced the cytotoxic effect of DNR in DNR-insensitive KG1a and Kasumi-1 cells, consistent with decreased Bcl-2 expression. Accordingly, siRNA against Bcl-2 increased the susceptibility of KG1a and Kasumi-1 cells to DNR-induced apoptosis. More importantly, curcumin suppressed Bcl-2 expression, selectively inhibited proliferation and synergistically enhanced the cytotoxicity of DNR in primary CD34+ AML cells, while showing limited lethality in normal CD34+ hematopoietic progenitors. Curcumin down-regulates Bcl-2 and induces apoptosis in DNR-insensitive CD34+ AML cell lines and primary CD34+ AML cells.
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6.5
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