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B 细胞淋巴瘤 2 卵巢杀伤因子（BOK）在氯化镉引起的人肺上皮细胞急性毒性中的作用。

Role of B-Cell Lymphoma 2 Ovarian Killer (BOK) in Acute Toxicity of Human Lung Epithelial Cells Caused by Cadmium Chloride.

机构信息

Institute for Chemical Carcinogenesis, Guangzhou Medical University, Guangzhou, Guangdong, China (mainland).

Institute of Toxicology, School of Public Health, Nanjing Medical University, Nanjing, Jiangsu, China (mainland).

出版信息

Med Sci Monit. 2019 Jul 19;25:5356-5368. doi: 10.12659/MSM.913706.

DOI:10.12659/MSM.913706
PMID:31323016
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6660808/
Abstract

BACKGROUND B-cell lymphoma 2 (BCL-2) ovarian killer (BOK) is a Bcl-2 family member with sequence homology to pro-apoptotic BAX and BAK, but its physiological and pathological roles remain largely unclear. Exposure of cells to cadmium may cause DNA damage, decrease DNA repair capacity, and increase genomic instability. MATERIAL AND METHODS The present study investigated the effects of BOK on the toxicity of cadmium chloride (CdCl₂) to human bronchial epithelial (16HBE) cells. We constructed BOK over-expressing (16HBE-BOK) cells and BOK knockdown (16HBE-shBOK) cells using the BOK-ORF plasmid and BOK-siRNA. qRT-PCR for BOK mRNA expression. We used Trypan blue exclusion assay for cell growth, MTT colorimetric assays for cells inhibition rate, and Comet assays for detecting damaged DNA. RESULTS CdCl₂, at various concentrations and exposure times, increased BOK mRNA. 16HBE-BOK cells (BOK over-expressing) proliferated more than 16HBE cells after 72 h; 16HBE-shBOK (BOK knockdown) cells proliferated less. In addition, BOK deficiency enhanced cell death induced by CdCl₂. Similarly, CdCl₂- and H₂O₂-induced DNA damage was greater in BOK-deficient cells. CONCLUSIONS These findings support a role for BOK in CdCl₂-induced DNA damage and cell death.

摘要

背景 B 细胞淋巴瘤 2（BCL-2）卵巢杀手（BOK）是 Bcl-2 家族成员，与促凋亡 BAX 和 BAK 具有序列同源性，但它的生理和病理作用在很大程度上仍不清楚。细胞暴露于镉可能导致 DNA 损伤、降低 DNA 修复能力并增加基因组不稳定性。

材料和方法 本研究探讨了 BOK 对氯化镉（CdCl₂）对人支气管上皮（16HBE）细胞毒性的影响。我们使用 BOK-ORF 质粒和 BOK-siRNA 构建了 BOK 过表达（16HBE-BOK）细胞和 BOK 敲低（16HBE-shBOK）细胞。qRT-PCR 检测 BOK mRNA 表达。我们使用台盼蓝排除试验检测细胞生长，MTT 比色法检测细胞抑制率，彗星试验检测受损 DNA。

结果 不同浓度和暴露时间的 CdCl₂均能增加 BOK mRNA。16HBE-BOK 细胞（BOK 过表达）在 72 小时后比 16HBE 细胞增殖更多；16HBE-shBOK（BOK 敲低）细胞增殖较少。此外，BOK 缺乏增强了 CdCl₂诱导的细胞死亡。同样，BOK 缺陷细胞中 CdCl₂和 H₂O₂诱导的 DNA 损伤更大。

结论 这些发现支持 BOK 在 CdCl₂诱导的 DNA 损伤和细胞死亡中的作用。

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