近几十年来,许多研究报道,在几种癌细胞系中,通过适当的照射,竹红菌素A(HA)可以消除癌细胞。然而,其抗癌作用的确切分子机制尚未完全确定。在光动力疗法(PDT)后评估人肺腺癌A549细胞中HA介导的细胞毒性和凋亡。引入了一种通过等量异位标签进行相对和绝对定量(iTRAQ)的时间定量蛋白质组学方法,该方法具有串联质谱(LC-MS/MS)的2D液相色谱,以帮助阐明分子细胞毒性机制并确定HA诱导的凋亡性细胞死亡的候选靶标。特异性半胱天冬酶抑制剂用于进一步阐明PDT处理的A549细胞中细胞凋亡的分子途径。最后,评估下游细胞凋亡相关蛋白。HA诱导的细胞凋亡与细胞收缩有关,细胞膜磷脂酰丝氨酸的外化,DNA片段化,和线粒体破坏,在此之前,细胞内活性氧(ROS)世代增加。进一步的研究表明,用0.08μmol/LHA进行PDT处理会导致线粒体破坏,细胞色素C的明显释放,和半胱天冬酶-3、-9和-7的激活。一起,HA可能是针对线粒体的可能治疗剂,也是有前途的光动力抗癌候选物,可用于进一步评估。
Over recent decades, many studies have reported that hypocrellin A (HA) can eliminate cancer cells with proper irradiation in several cancer cell lines. However, the precise molecular mechanism underlying its anticancer effect has not been fully defined. HA-mediated cytotoxicity and apoptosis in human lung adenocarcinoma A549 cells were evaluated after photodynamic therapy (PDT). A temporal quantitative proteomics approach by isobaric tag for relative and absolute quantitation (iTRAQ) 2D liquid chromatography with tandem mass spectrometric (LC-MS/MS) was introduced to help clarify molecular cytotoxic mechanisms and identify candidate targets of HA-induced apoptotic cell death. Specific caspase inhibitors were used to further elucidate the molecular pathway underlying apoptosis in PDT-treated A549 cells. Finally, down-stream apoptosis-related protein was evaluated. Apoptosis induced by HA was associated with cell shrinkage, externalization of cell membrane phosphatidylserine, DNA fragmentation, and mitochondrial disruption, which were preceded by increased intracellular reactive oxygen species (ROS) generations. Further studies showed that PDT treatment with 0.08 µmol/L HA resulted in mitochondrial disruption, pronounced release of cytochrome c, and activation of caspase-3, -9, and -7. Together, HA may be a possible therapeutic agent directed toward mitochondria and a promising photodynamic anticancer candidate for further evaluation.