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19 July 2019 : Laboratory Research  

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

Fang Zhang1ABCDEF, Liang Ren1ABCD, Shanshan Zhou1ABCD, Peng Duan1BC, Junchao Xue2BC, Haiqin Chen1BC, Yufeng Feng1BC, Xiaoxuan Yue1BC, Piaofan Yuan1BC, Qizhan Liu2E, Ping Yang1ABCDEFG*, Yixiong Lei1ABCDEFG

DOI: 10.12659/MSM.913706

Med Sci Monit 2019; 25:5356-5368

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.

Keywords: bcl-Associated Death Protein, Cadmium Chloride, Comet Assay, Bronchi, Cell Death, DNA Damage, Proto-Oncogene Proteins c-bcl-2, RNA, Messenger

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Medical Science Monitor eISSN: 1643-3750
Medical Science Monitor eISSN: 1643-3750