Ashwaq Al-Abboodi Shakir, Al-Qubaisi Mothanna Sadiq, Rasedee Abdullah, Abdul Ahmad Bustamam, Taufiq-Yap Yun Hin, Yeap Swee Keong
MAKNA-UPM, Cancer Research Laboratory, Institute of Bioscience, University Putra Malaysia, 43400 Selangor, Malaysia.
Department of Veterinary Laboratory Diagnosis, Faculty of Veterinary Medicine, University Putra Malaysia, 43400 Selangor, Malaysia.
Int J Mol Sci. 2016 Oct 18;17(10):1653. doi: 10.3390/ijms17101653.
Dentatin (DEN), purified from the roots of Burm f., has poor aqueous solubility that reduces its therapeutic application. The aim of this study was to assess the effects of DEN-HPβCD (hydroxypropyl-β-cyclodextrin) complex as an anticancer agent in HT29 cancer cell line and compare with a crystal DEN in dimethyl sulfoxide (DMSO). The exposure of the cancer cells to DEN or DEN-HPβCD complex leads to cell growth inhibition as determined by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. To analyze the mechanism, in which DEN or DEN-HPβCD complex causes the death in human colon HT29 cancer cells, was evaluated by the enzyme-linked immunosorbent assay (ELIZA)-based assays for caspase-3, 8, 9, and reactive oxygen species (ROS). The findings showed that an anti-proliferative effect of DEN or DEN-HPβCD complex were via cell cycle arrest at the G2/M phase and eventually induced apoptosis through both mitochondrial and extrinsic pathways. The down-regulation of poly(ADP-ribose) polymerase (PARP) which leaded to apoptosis upon treatment, was investigated by Western-blotting. Hence, complexation between DEN and HPβCD did not diminish or eliminate the effective properties of DEN as anticancer agent. Therefore, it would be possible to resolve the conventional and current issues associated with the development and commercialization of antineoplastic agents in the future.
从缅甸紫铆根中纯化得到的齿孔酸（DEN），其水溶性较差，这限制了它的治疗应用。本研究的目的是评估齿孔酸 - 羟丙基 -β-环糊精（HPβCD）复合物作为抗癌剂对HT29癌细胞系的作用，并与二甲基亚砜（DMSO）中的结晶齿孔酸进行比较。通过MTT（3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐）试验确定，癌细胞暴露于齿孔酸或齿孔酸 - HPβCD复合物会导致细胞生长抑制。为了分析齿孔酸或齿孔酸 - HPβCD复合物导致人结肠HT29癌细胞死亡的机制，通过基于酶联免疫吸附测定（ELISA）的方法检测了半胱天冬酶 - 3、8、9和活性氧（ROS）。研究结果表明，齿孔酸或齿孔酸 - HPβCD复合物的抗增殖作用是通过使细胞周期停滞在G2/M期，并最终通过线粒体和外源性途径诱导细胞凋亡。通过蛋白质免疫印迹法研究了聚（ADP - 核糖）聚合酶（PARP）的下调情况，该酶在治疗后会导致细胞凋亡。因此，齿孔酸与HPβCD的络合并没有削弱或消除齿孔酸作为抗癌剂的有效特性。所以，未来有可能解决与抗肿瘤药物开发和商业化相关的传统和当前问题。
