Sun Weimin, Wang Wei, Kim Jung, Keng Peter, Yang Shanmin, Zhang Hengshan, Liu Chaomei, Okunieff Paul, Zhang Lurong
Department of Radiation Oncology, University of Rochester Medical Center, Rochester, NY 14642, USA.
Adv Exp Med Biol. 2008;614:179-86. doi: 10.1007/978-0-387-74911-2_21.
Resveratrol, a phytoalexin found in the skin of grapes, is believed to have multiple bioactivities including anti-cancer, anti-carcinogenesis and antiinflammatory. The mechanisms by which resveratrol might produce these effects are not well understood. In this study, malignant human pancreatic cancer cells were treated without or with resveratrol in combination with ionizing radiation (IR), and then the mitochondrial function of treated cells was evaluated using several standardized assays. They include the Calcein AM method for mitochondria transition pore; the JC-1 staining method for mitochondria membrane potential; the CM-H2DCFDA method for reactive oxygen species; and the Annexin V/propidium iodide (PI) method for apoptosis/cell death. Our results indicated that (1) pore function was partially intact after resveratrol, but resveratrol probably interfered with the accumulation of intracellular Calcein AM; (2) depolarization of the mitochondria membrane was increased in the resveratrol treated cells, consistent with mitochondrial dysfunction; (3) ROS was slightly increased with resveratrol, a phenomenon that was greatly increased when this agent was combined with IR; and (4) in parallel with the above changes in mitochondrial and drug transport, cells treated with resveratrol showed increased apoptosis as measured by Annexin V/PI staining. In summary, the anti-cancer effect of resveratrol is associated with the damage of mitochondrial function that leads to increased ROS, apoptosis, and possibly intracellular drug accumulation via inhibition of proteins involved in multi-drug resistance (MDR).
白藜芦醇是一种存在于葡萄皮中的植物抗毒素，被认为具有多种生物活性，包括抗癌、抗致癌作用和抗炎作用。白藜芦醇产生这些效应的机制尚未完全明确。在本研究中，对恶性人胰腺癌细胞进行处理，一组不使用白藜芦醇，另一组使用白藜芦醇并联合电离辐射（IR），然后使用几种标准化检测方法评估处理后细胞的线粒体功能。这些检测方法包括用于线粒体通透性转换孔的钙黄绿素乙酰甲酯（Calcein AM）法；用于线粒体膜电位的JC-1染色法；用于活性氧的CM-H2DCFDA法；以及用于凋亡/细胞死亡的膜联蛋白V/碘化丙啶（PI）法。我们的结果表明：（1）白藜芦醇处理后孔功能部分完整，但白藜芦醇可能干扰了细胞内钙黄绿素乙酰甲酯的积累；（2）白藜芦醇处理的细胞中线粒体膜去极化增加，这与线粒体功能障碍一致；（3）白藜芦醇使活性氧略有增加，当该药物与电离辐射联合使用时，这种现象显著增加；（4）与线粒体和药物转运的上述变化同时，通过膜联蛋白V/PI染色检测，白藜芦醇处理的细胞凋亡增加。总之，白藜芦醇的抗癌作用与线粒体功能损伤有关，线粒体功能损伤导致活性氧增加、细胞凋亡，并可能通过抑制参与多药耐药（MDR）的蛋白质导致细胞内药物积累。
