咖啡中的天然二萜类化合物——咖啡醇和卡韦醇通过调节人恶性胸膜间皮瘤中特异性蛋白 1 的表达诱导细胞凋亡。
Natural diterpenes from coffee, cafestol and kahweol induce apoptosis through regulation of specificity protein 1 expression in human malignant pleural mesothelioma.
机构信息
Department of Biochemistry, College of Medicine, Soonchunhyang University, Ssangyong-dong, Seobuk-gu, Cheonan, Choongnam 331-090, Republic of Korea.
出版信息
J Biomed Sci. 2012 Jun 26;19(1):60. doi: 10.1186/1423-0127-19-60.
BACKGROUND
Malignant pleural mesothelioma (MPM) is a highly aggressive cancer with a very poor prognosis. Several clinical studies such as immunotherapy, gene therapy and molecular targeting agents have been tried for treatment of malignant mesothelioma, however, there is no application for effective clinical treatment. Coffee has various biological functions such as anti-oxidant, anti-inflammatory, anti-mutagenic and anti-carcinogenic activities. The therapeutic activities of the bioactive compounds in coffee was sugested to influence intracellular signaling of MPM. Regarding to the cancer-related functions, In this study, suppression of Sp1 protein level followed by induction of MSTO-211H cell apoptosis by cafestol and kahweol were investigated in oreder to determine Sp1's potential as a significant target for human MPM therapy as well.
METHODS
Cells were treated separately with final concentration of cafestol and kahweol and the results were analyzed by MTS assay, DAPI staining, PI staining, luciferase assay, RT-PCR, and immunoblotting.
RESULTS
Viability of MSTO-211H and H28 cells were decreased, and apoptotic cell death was increased in MSTO-211H as a result of cafestol and kahweol treatment. Cafestol and kahweol increased Sub-G1 population and nuclear condensation in MSTO-211H cells. Roles of Sp1 in cell proliferation and apoptosis of the MSTO-211H cells by the Sp1 inhibitor of Mithramycin A were previously confirmed. Cafestol and kahweol significantly suppressed Sp1 protein levels. Kahweol slightly attenuated Sp1 mRNA, while Cafestol did not affect in MSTO-211H cells. Cafestol and kahweol modulated the promoter activity and protein expression level of the Sp1 regulatory genes including Cyclin D1, Mcl-1, and Survivin in mesothelioma cells. Apoptosis signaling cascade was activated by cleavages of Bid, Caspase-3, and PARP with cafestol and by upregulation of Bax, and downregulation of Bcl-xl by kahweol.
CONCLUSIONS
Sp1 can be a novel molecular target of cafestol and kahweol in human MPM.
背景
恶性胸膜间皮瘤（MPM）是一种侵袭性很强的癌症，预后极差。免疫疗法、基因疗法和分子靶向药物等多种临床研究已尝试用于治疗恶性间皮瘤，但尚无有效的临床治疗方法。咖啡具有抗氧化、抗炎、抗突变和抗癌等多种生物学功能。咖啡中生物活性化合物的治疗活性被认为影响 MPM 的细胞内信号。关于癌症相关功能，在这项研究中，通过咖啡醇和卡瓦醇抑制 Sp1 蛋白水平并诱导 MSTO-211H 细胞凋亡，以确定 Sp1 作为人类 MPM 治疗的重要靶点的潜力。
方法
分别用咖啡醇和卡瓦醇的终浓度处理细胞，通过 MTS 分析、DAPI 染色、PI 染色、荧光素酶分析、RT-PCR 和免疫印迹分析结果。
结果
MSTO-211H 和 H28 细胞的活力降低，MSTO-211H 细胞的凋亡细胞死亡增加，这是由于咖啡醇和卡瓦醇的处理。咖啡醇和卡瓦醇增加了 MSTO-211H 细胞的 Sub-G1 群体和核浓缩。先前已证实 Sp1 抑制剂米托蒽醌 A 在 MSTO-211H 细胞中对细胞增殖和细胞凋亡的作用。咖啡醇和卡瓦醇显著抑制 Sp1 蛋白水平。卡瓦醇轻微降低 Sp1 mRNA，而咖啡醇在 MSTO-211H 细胞中没有影响。咖啡醇和卡瓦醇调节了 Sp1 调节基因的启动子活性和蛋白表达水平，包括间皮瘤细胞中的细胞周期蛋白 D1、Mcl-1 和 Survivin。咖啡醇和卡瓦醇通过 Bid、Caspase-3 和 PARP 的裂解和通过 Bax 的上调以及 Bcl-xl 的下调激活了凋亡信号级联。
结论
Sp1 可成为咖啡醇和卡瓦醇在人 MPM 中的新型分子靶标。
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