Skipper Anthony, Sims Jennifer N, Yedjou Clement G, Tchounwou Paul B
Molecular Toxicology Research Laboratory, NIH-Center for Environmental Health, College of Science, Engineering and Technology, Jackson State University, 1400 Lynch Street, Box 18540, Jackson, MS 39217, USA.
Department of Pathology, Beth Israel Deaconess Medical Center, Harvard Medical School, 330 Bookline Avenue, Boston, MA 02215, USA.
Int J Environ Res Public Health. 2016 Jan 2;13(1):88. doi: 10.3390/ijerph13010088.
Cadmium is a heavy metal that has been shown to cause its toxicity in humans and animals. Many documented studies have shown that cadmium produces various genotoxic effects such as DNA damage and chromosomal aberrations. Ailments such as bone disease, renal damage, and several forms of cancer are attributed to overexposure to cadmium. Although there have been numerous studies examining the effects of cadmium in animal models and a few case studies involving communities where cadmium contamination has occurred, its molecular mechanisms of action are not fully elucidated. In this research, we hypothesized that oxidative stress plays a key role in cadmium chloride-induced toxicity, DNA damage, and apoptosis of human liver carcinoma (HepG₂) cells. To test our hypothesis, cell viability was determined by MTT assay. Lipid hydroperoxide content stress was estimated by lipid peroxidation assay. Genotoxic damage was tested by the means of alkaline single cell gel electrophoresis (Comet) assay. Cell apoptosis was measured by flow cytometry assessment (Annexin-V/PI assay). The result of MTT assay indicated that cadmium chloride induces toxicity to HepG₂ cells in a concentration-dependent manner, showing a 48 hr-LD50 of 3.6 µg/mL. Data generated from lipid peroxidation assay resulted in a significant (p < 0.05) increase of hydroperoxide production, specifically at the highest concentration tested. Data obtained from the Comet assay indicated that cadmium chloride causes DNA damage in HepG₂ cells in a concentration-dependent manner. A strong concentration-response relationship (p < 0.05) was recorded between annexin V positive cells and cadmium chloride exposure. In summary, these in vitro studies provide clear evidence that cadmium chloride induces oxidative stress, DNA damage, and programmed cell death in human liver carcinoma (HepG₂) cells.
镉是一种重金属,已被证明会对人类和动物产生毒性。许多有文献记载的研究表明,镉会产生各种遗传毒性效应,如DNA损伤和染色体畸变。诸如骨病、肾损伤和几种癌症等疾病都归因于镉的过度暴露。尽管已经有大量研究考察了镉在动物模型中的作用,以及一些涉及镉污染社区的案例研究,但其分子作用机制尚未完全阐明。在本研究中,我们假设氧化应激在氯化镉诱导的人肝癌(HepG₂)细胞毒性、DNA损伤和细胞凋亡中起关键作用。为了验证我们的假设,通过MTT法测定细胞活力。通过脂质过氧化测定法评估脂质过氧化氢含量应激。通过碱性单细胞凝胶电泳(彗星)测定法检测遗传毒性损伤。通过流式细胞术评估(膜联蛋白V/碘化丙啶测定法)测量细胞凋亡。MTT法的结果表明,氯化镉以浓度依赖性方式诱导对HepG₂细胞的毒性,显示48小时半数致死剂量为3.6μg/mL。脂质过氧化测定法产生的数据导致过氧化氢产生显著增加(p<0.05),特别是在测试的最高浓度下。从彗星试验获得的数据表明,氯化镉以浓度依赖性方式导致HepG₂细胞中的DNA损伤。在膜联蛋白V阳性细胞与氯化镉暴露之间记录到强烈的浓度-反应关系(p<0.05)。总之,这些体外研究提供了明确的证据,表明氯化镉在人肝癌(HepG₂)细胞中诱导氧化应激、DNA损伤和程序性细胞死亡。