Ali Norlaily Mohd, Yeap Swee Keong, Abu Nadiah, Lim Kian Lam, Ky Huynh, Pauzi Ahmad Zaim Mat, Ho Wan Yong, Tan Sheau Wei, Alan-Ong Han Kiat, Zareen Seema, Alitheen Noorjahan Banu, Akhtar M Nadeem
Faculty of Medicine and Health Sciences, Universiti Tunku Abdul Rahman, Lot PT 21144, Jalan Sungai Long, Bandar Sungai Long, Cheras, 43000 Kajang, Selangor Malaysia.
China-ASEAN College of Marine Sciences, Xiamen University Malaysia, Jalan Sunsuria, Bandar Sunsuria, 43900 Sepang, Selangor Malaysia.
Cancer Cell Int. 2017 Feb 21;17:30. doi: 10.1186/s12935-017-0400-3. eCollection 2017.
Curcumin is a lead compound of the rhizomes of and possess a broad range of pharmacological activities. Chemically, curcumin is 1,3-dicarbonyl class of compound, which exhibits keto-enol tautomerism. Despite of its strong biological properties, curcumin has yet been recommended as a therapeutic agent because of its poor bioavailability.
A curcumin derivative ()-3-hydroxy-1-(2-hydroxyphenyl)-3-phenylprop-2-en-1-one (DK1) was synthesized and its cytotoxicity was tested on breast cancer cell MCF-7 and normal cell MCF-10A using MTT assay. Meanwhile, cell cycle regulation and apoptosis on MCF-7 cell were evaluated using flow cytometry. Regulation of cell cycle and apoptosis related genes expression was investigated by quantitative real time polymerase chain reaction (qRT-PCR), western blot and caspases activity analyses. Activation of oxidative stress on MCF-7 were evaluated by measuring ROS and GSH levels.
DK1 was found to possess selective cytotoxicity on breast cancer MCF-7 cell than normal MCF-10A cell. Flow cytometry cell cycle and AnnexinV/PI analyses reported that DK1 effectively arrested MCF-7 at G2/M phase and induced apoptosis after 72 h of incubation than curcumin. Upregulation of p53, p21 and downregulation of PLK-1 subsequently promote phosphorylation of CDC2 which were found contributed to the arrest of G2/M phase. Moreover, increased of reactive oxygen species and reduced of antioxidant glutathione level correlate with apoptosis observed with raised of cytochrome c and active caspase 9.
DK1 was found to be more effective in inducing cell cycle arrest and apoptosis against MCF-7 cell with much higher selectivity index of MCF-10A/MCF-7 than curcumin, which might be contributed by the overexpression of p53 protein.
姜黄素是姜黄根茎的主要成分，具有广泛的药理活性。从化学结构上看，姜黄素属于1,3 - 二羰基类化合物，存在酮 - 烯醇互变异构现象。尽管姜黄素具有强大的生物学特性，但由于其生物利用度差，尚未被推荐作为治疗药物。
合成了一种姜黄素衍生物（）-3 - 羟基 - 1 -（2 - 羟基苯基）-3 - 苯基丙 - 2 - 烯 - 1 - 酮（DK1），并使用MTT法检测其对乳腺癌细胞MCF - 7和正常细胞MCF - 10A的细胞毒性。同时，采用流式细胞术评估MCF - 7细胞的细胞周期调控和凋亡情况。通过定量实时聚合酶链反应（qRT - PCR）、蛋白质印迹法和半胱天冬酶活性分析研究细胞周期和凋亡相关基因表达的调控。通过测量活性氧（ROS）和谷胱甘肽（GSH）水平评估MCF - 7细胞氧化应激的激活情况。
发现DK1对乳腺癌MCF - 7细胞具有比正常MCF - 10A细胞更高的选择性细胞毒性。流式细胞术细胞周期和膜联蛋白V/碘化丙啶（AnnexinV/PI）分析表明，与姜黄素相比，DK1在孵育72小时后能有效将MCF - 7细胞阻滞在G2/M期并诱导凋亡。p53、p21的上调以及PLK - 1的下调随后促进了细胞周期蛋白依赖性激酶2（CDC2）的磷酸化，这被认为与G2/M期的阻滞有关。此外，活性氧的增加和抗氧化剂谷胱甘肽水平的降低与凋亡相关，同时观察到细胞色素c和活性半胱天冬酶9的增加。
发现DK1在诱导MCF - 7细胞周期阻滞和凋亡方面比姜黄素更有效，其MCF - 10A/MCF - 7选择性指数更高，这可能是由于p53蛋白的过表达所致。
