Hassan Reem N, Luo Hualei, Jiang Weiying
Department of Medical Genetics, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou, Guangdong 510080, People's Republic of China.
Cancer Manag Res. 2020 Feb 12;12:1089-1100. doi: 10.2147/CMAR.S229395. eCollection 2020.
The present study aimed to examine the effects of nicotinamide (NAM) on cervical cancer-associated fibroblasts (CAF) for its in vitro efficacy, gross inhibition, and mechanism of inhibition.
The fibroblasts were treated with pre-specified concentrations of NAM followed by measurement of the cell proliferation using CCK-8 assay. The production of reactive oxygen species (ROS) was measured by 2',7'-Dichlorofluorescin diacetate. We further investigated the apoptosis by flow cytometry using Annexin-V. We employed JC-1 assay to detect changes in the potential of the mitochondrial membrane. We further determined the expression of apoptotic genes was measured using qRT-PCR. And lastly, cell cycle experiments were conducted to determine the influence of NAM on arresting the growth of CAF in a cell cycle.
Our study showed that NAM was able to reduce fibroblasts viability. We specifically observed a significantly increased intracellular ROS with resultant exhaustion of cellular antioxidant defense machinery, including reduced glutathione (GSH). We further observed the involvement of mitochondrial pathway in the NAM induced apoptosis of fibroblasts.
Our study supports the therapeutic potential of NAM for the treatment of cervical cancer and necessitates a further investigation of the reported findings.
本研究旨在探讨烟酰胺（NAM）对宫颈癌相关成纤维细胞（CAF）的体外疗效、总体抑制作用及抑制机制。
用预先设定浓度的NAM处理成纤维细胞，然后使用CCK-8法测量细胞增殖。用二氯二氢荧光素二乙酸酯测定活性氧（ROS）的产生。我们进一步使用膜联蛋白-V通过流式细胞术研究细胞凋亡。我们采用JC-1法检测线粒体膜电位的变化。我们进一步使用qRT-PCR测定凋亡基因的表达。最后，进行细胞周期实验以确定NAM对细胞周期中CAF生长停滞的影响。
我们的研究表明，NAM能够降低成纤维细胞的活力。我们特别观察到细胞内ROS显著增加，导致细胞抗氧化防御机制耗尽，包括谷胱甘肽（GSH）减少。我们进一步观察到线粒体途径参与了NAM诱导的成纤维细胞凋亡。
我们的研究支持NAM治疗宫颈癌的潜在疗效，需要对报道的结果进行进一步研究。
