Nucleus of Multidisciplinary Research in Biology (Numpex-Bio), Universidade Federal Do Rio de Janeiro (UFRJ), Rio de Janeiro, Duque de Caxias, Brazil.
Mol Biol Rep. 2021 Jun;48(6):5199-5207. doi: 10.1007/s11033-021-06524-5. Epub 2021 Jul 1.
The 2,3,5-triiodobenzoic acid (TIBA) is an iodine contrast agent used for visualization of tissue in X-ray techniques. However, TIBA induces physiological complications like increase in oxygen reactive species (ROS), and consequently, contrast-induced nephropathies. TIBA's antitumor activity was demonstrated in lung cancer, but the subcellular mechanisms involving its activity in tumor cells are still unknown. Thus, the objective of this work was evaluate whether the anti-tumor activity of TIBA involves ROS increase, in tumor lines of non-small cell lung cancer (H460), chronic myeloid leukemia (K562), and its cytotoxicity in normal renal epithelial (VERO). The MTT (3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide) assay was used for evaluation of cell viability, the HDCFDA (cell-permeant 2',7'-dichlorodihydrofluorescein diacetate) fluorescent probe to evaluate ROS induction, cell cycle analysis was performed using flow cytometry to measure cell death, and immunofluorescence with annexin/7-AAD (7-amino-actinomycin D), to assess the association of cell death with the ROS generation. TIBA decreases cell viability in a dose-dependent manner for the H460 and K562. However, VERO cells showed less response to the drug, with 70% viable cells after 72 h of treatment in the highest concentration of the drug. While the tumor cells with only 20% viable cells. Besides, tumor cells exhibited higher DNA fragmentation, compared to the renal line (VERO with 5% of fragmented DNA, H460 with 26%, and 56% in K562). Finally, TIBA-induced ROS increase and apoptosis in all lines, which is significantly decreased after treatment with the antioxidant N-acetyl-cysteine (NAC). These data demonstrate the relationship between the increased cellular oxidative stress and the anti-tumor action of the TIBA.
2,3,5-三碘苯甲酸（TIBA）是一种碘造影剂，用于 X 射线技术中组织的可视化。然而，TIBA 会引起生理并发症，如氧活性物质（ROS）增加，进而导致对比剂肾病。TIBA 在肺癌中表现出抗肿瘤活性，但涉及其在肿瘤细胞中活性的亚细胞机制尚不清楚。因此，本工作的目的是评估 TIBA 的抗肿瘤活性是否涉及 ROS 增加，在非小细胞肺癌（H460）、慢性髓性白血病（K562）肿瘤细胞系及其在正常肾上皮（VERO）中的细胞毒性。MTT（3-（4,5-二甲基噻唑-2-基）-2,5-二苯基四氮唑溴盐）测定法用于评估细胞活力，HDCFDA（细胞通透 2',7'-二氯二氢荧光素二乙酸酯）荧光探针用于评估 ROS 诱导，用流式细胞术进行细胞周期分析以测量细胞死亡，并用 Annexin/7-AAD（7-氨基放线菌素 D）进行免疫荧光染色，以评估细胞死亡与 ROS 生成的关联。TIBA 以剂量依赖的方式降低 H460 和 K562 细胞的活力。然而，VERO 细胞对药物的反应较小，在药物的最高浓度下，72 小时后仍有 70%的活细胞。而肿瘤细胞仅有 20%的活细胞。此外，与肾系（VERO 有 5%的 DNA 片段化，H460 有 26%，K562 有 56%）相比，肿瘤细胞表现出更高的 DNA 片段化。最后，TIBA 诱导所有细胞系中的 ROS 增加和细胞凋亡，在用抗氧化剂 N-乙酰半胱氨酸（NAC）处理后，这种增加显著减少。这些数据表明，细胞氧化应激增加与 TIBA 的抗肿瘤作用之间存在关系。
