Laboratory of Biochemistry and Immunology, College of Veterinary Medicine, Chungbuk National University, Cheongju, Chungbuk, Republic of Korea.
College of Pharmacy, Institute of Pharmaceutical Science and Technology, Hanyang University, Ansan, Republic of Korea.
Toxicol In Vitro. 2018 Oct;52:161-169. doi: 10.1016/j.tiv.2018.06.007. Epub 2018 Jun 9.
Cigarette smoke contains thousands of harmful components, many of which exert toxic effects on reproductive organs, including the ovaries, and their development. In this study, we investigated the effects of cigarette smoke extract (CSE) on cell proliferation, apoptosis and oxidative stress in the SKOV3 and OVCAR3 ovarian cancer cell lines. A water soluble tetrazolium (WST) assay revealed that the cell proliferation of both ovarian cells was significantly inhibited by three types of CSE in a concentration-dependent manner. Western blot analysis showed that the inhibition of cell proliferation was related to regulation of cell cycle-related protein expression. To determine the effects of cigarette smoke on the death of ovarian cells, we conducted a TUNEL assay, which revealed a time-dependent increase in TUNEL-stained cells in response to CSE exposure. Moreover, the protein expression of the Bcl-2 family signaling pathway observed upon Western blot analysis were consistent with the results of the TUNEL assay. We also examined the reactive oxygen species (ROS) generation in two ovarian cell lines using DCF-DA (5(6)-Carboxy-2',7'-dichlorofluorescein diacetate) assay and ROS-Glo™ HO assay to investigate the oxidative stress caused by CSE. Both ROS detection assays demonstrated an increase in the amount of ROS by CSE in ovarian cells, which was consistent with the results observed for the Keap1-Nrf2 pathway upon Western blot analysis. Taken together, these results suggest that CSE may induce cell proliferation inhibition and oxidative stress in ovarian cells.
香烟烟雾中含有数千种有害成分，其中许多成分对生殖器官（包括卵巢）及其发育具有毒性作用。在这项研究中，我们研究了香烟烟雾提取物（CSE）对 SKOV3 和 OVCAR3 卵巢癌细胞系细胞增殖、凋亡和氧化应激的影响。水溶性四唑盐（WST）检测结果显示，三种 CSE 以浓度依赖的方式显著抑制了两种卵巢细胞的增殖。Western blot 分析表明，细胞增殖的抑制与细胞周期相关蛋白表达的调节有关。为了确定香烟烟雾对卵巢细胞死亡的影响，我们进行了 TUNEL 检测，结果显示随着 CSE 暴露，TUNEL 染色细胞的数量呈时间依赖性增加。此外，Western blot 分析观察到的 Bcl-2 家族信号通路蛋白表达与 TUNEL 检测结果一致。我们还使用 DCF-DA（5(6)-羧基-2',7'-二氯荧光素二乙酸酯）检测和 ROS-Glo™ HO 检测两种卵巢细胞系中的活性氧（ROS）生成，以研究 CSE 引起的氧化应激。两种 ROS 检测均表明 CSE 增加了卵巢细胞中的 ROS 含量，这与 Western blot 分析中观察到的 Keap1-Nrf2 通路结果一致。综上所述，这些结果表明 CSE 可能诱导卵巢细胞增殖抑制和氧化应激。
