Laboratory of Pharmacogenetics, Drug Research and Development Center (NPDM), Federal University of Ceará, Cel. Nunes de Melo, 1000 - Rodolfo Teófilo, Fortaleza, Brazil; Laboratory of Human Cytogenetics, Institute of Biological Sciences, Federal University of Pará, Augusto Correa Avenue, 01 - Guamá, Belém, Brazil.
Laboratory of Pharmacogenetics, Drug Research and Development Center (NPDM), Federal University of Ceará, Cel. Nunes de Melo, 1000 - Rodolfo Teófilo, Fortaleza, Brazil.
Toxicol Appl Pharmacol. 2022 Dec 1;456:116256. doi: 10.1016/j.taap.2022.116256. Epub 2022 Oct 5.
Colorectal cancer (CRC) is estimated as the third most incident cancer and second in mortality worldwide. Moreover, CRC metastasis reduces patients' survival rates. Thus, the study and identification of new compounds with anticancer activity selectively to tumor cells are encouraged in the CRC treatment. Naphtoquinones are compounds with several pharmacologic activities, including antitumoral properties. Therefore, this study aimed to investigate the anticancer mechanism of synthetic 8-Hydroxy-2-(P-Nitrothiophenol)-1,4-Naphthoquinone (CNN16) in colon cancer cell line HCT-116. CNN16 showed an IC of 5.32 μM in HCT-116, and 9.36, 10.77, and 24.57 μM in the non-cancerous cells MRC-5, MNP-01, and PMBC, respectively, evaluated by the MTT assay. CNN16 showed an anticlonogenic effect in HCT-116 and induced cell fragmentation identified by flow cytometry analysis. Furthermore, we observed that CNN16 presented genotoxicity and induces reactive oxygen species (ROS) after 3 h of treatment visualized by alkaline comet assay and DCFH-DA dye fluorescence, respectively. Furthermore, CNN16 caused cellular membrane disruption, reduction in the mitochondrial membrane polarization, and the presence of apoptotic bodies and chromatin condensation was visualized by differential stained (HO/FD/PI) in fluorescent microscopy along with PARP1, TP53, BCL-2, and BAX analyzed by RT-qPCR. Results also evidenced inhibition in the migratory process analyzed by wound healing assay. Therefore, CNN16 can be considered as a potential new leader molecule for CRC treatment, although further studies are still necessary to comprehend the effects of CNN16 in in vivo models to evaluate the anti-migratory effect, and toxicology and assure compound safety and selectively.
结直肠癌（CRC）是全球发病率第三、死亡率第二的癌症。此外，CRC 的转移降低了患者的生存率。因此，鼓励在 CRC 治疗中研究和鉴定具有抗癌活性且对肿瘤细胞具有选择性的新化合物。萘醌类化合物具有多种药理活性，包括抗肿瘤特性。因此，本研究旨在探讨合成 8-羟基-2-（对硝基硫酚）-1,4-萘醌（CNN16）在结肠癌细胞系 HCT-116 中的抗癌机制。MTT 测定法评估，CNN16 在 HCT-116 中的 IC 为 5.32μM，而在非癌细胞 MRC-5、MNP-01 和 PMBC 中的 IC 分别为 9.36、10.77 和 24.57μM。CNN16 在 HCT-116 中表现出抗集落形成作用，并通过流式细胞术分析鉴定细胞碎片。此外，我们观察到 CNN16 在 3 小时的处理后具有遗传毒性，并通过碱性彗星试验和 DCFH-DA 染料荧光分别诱导活性氧（ROS）的产生。此外，CNN16 引起细胞膜破裂、线粒体膜去极化减少，并通过荧光显微镜观察到凋亡小体和染色质浓缩，并用差异染色（HO/FD/PI），同时通过 RT-qPCR 分析 PARP1、TP53、BCL-2 和 BAX。结果还证明了划痕愈合试验中迁移过程的抑制。因此，CNN16 可以被认为是 CRC 治疗的一种有潜力的新先导分子，尽管仍需要进一步研究以了解 CNN16 在体内模型中的作用，以评估其抗迁移作用、毒理学和确保化合物的安全性和选择性。
