Laboratory of Biochemistry and Immunology, College of Veterinary Medicine, Chungbuk National University, Korea.
J Toxicol Sci. 2020;45(11):713-724. doi: 10.2131/jts.45.713.
Acrylonitrile (AN), which is widely utilized in the manufacture of plastics, acrylamide, acrylic fibers, and resins, is also one of main components of cigarette smoke (CS). In this study, we examined the effects of AN on the cell viability and apoptosis of JEG-3 and BeWo human choriocarcinoma cancer cell lines. A cell viability assay confirmed that AN decreased the cell proliferation of JEG-3 and BeWo cells in a dose-dependent manner. Additionally, Western blot assay revealed that protein expression of cyclin D and cyclin E decreased, while protein expression of p21 and p27 increased in response to AN treatment for 48 hr. The changes in reactive oxygen species (ROS) levels in JEG-3 and BeWo cells exposed to AN were also measured by a dichlorofluorescein diacetate (DCFH-DA) assay, which revealed that ROS levels increased in response to AN treatment for 48 hr. Moreover, western blot assay confirmed that AN treatment of JEG-3 and BeWo cells for 4 hr promoted the expression of phosphorylated eukaryotic initiation factor 2 alpha protein (p-eIF2α), C/EBP homologous protein (CHOP) and caspase 12, which are known to be involved in ROS-mediated endoplasmic reticulum stress (ER-stress)-related apoptosis. Overall, the protein expression of p53 and Bax (a pro-apoptosis marker) increased, while the expression of Bcl-xl (an anti-apoptotic marker) decreased and the number of apoptotic cells increased in response to AN treatment for 48 hr. Taken together, these results suggest that AN has the potential to induce apoptosis of JEG-3 and BeWo human choriocarcinoma cancer cells by activating ROS.
丙烯腈（AN）广泛用于塑料、丙烯酰胺、丙烯酸纤维和树脂的制造，也是香烟烟雾（CS）的主要成分之一。在这项研究中，我们研究了丙烯腈对 JEG-3 和 BeWo 人绒毛膜癌细胞系细胞活力和细胞凋亡的影响。细胞活力测定证实，丙烯腈以剂量依赖的方式降低了 JEG-3 和 BeWo 细胞的增殖。此外，Western blot 分析显示，cyclin D 和 cyclin E 的蛋白表达减少，而 p21 和 p27 的蛋白表达增加，这是对丙烯腈处理 48 小时的反应。通过二氯荧光素二乙酸酯（DCFH-DA）测定还测量了 JEG-3 和 BeWo 细胞中丙烯腈暴露后活性氧（ROS）水平的变化，结果表明 ROS 水平随着丙烯腈处理 48 小时而增加。此外，Western blot 分析证实，JEG-3 和 BeWo 细胞用丙烯腈处理 4 小时后，促进了磷酸化真核起始因子 2 阿尔法蛋白（p-eIF2α）、C/EBP 同源蛋白（CHOP）和半胱天冬酶 12 的表达，这些蛋白已知参与 ROS 介导的内质网应激（ER-stress）相关凋亡。总的来说，p53 和 Bax（促凋亡标志物）的蛋白表达增加，而 Bcl-xl（抗凋亡标志物）的表达减少，凋亡细胞的数量在丙烯腈处理 48 小时后增加。综上所述，这些结果表明丙烯腈通过激活 ROS 有可能诱导 JEG-3 和 BeWo 人绒毛膜癌细胞的凋亡。
