采用基于γ-H2AX焦点分析的流式细胞术检测纳米金颗粒的遗传毒性

doi:10.3969/j.issn.1004-616x.2024.05.012

Abstract

Abstract: OBJECTIVE： To use a flow cytometry genotoxicity assay based on γ-H2AX biomarker for improvement of genotoxicity evaluation of gold nanoparticles. METHODS： Final concentrations of 7.5，3.75，and 1.875 μg/mL of Cyclophosphamide were used as the positive controls for establishment of the γ-H2AX assay in +S₉ group (4 hours of short treatment) and final concentrations of 6.4，1.6，and 0.4 μg/mL of Etoposide were used as the positive controls for the establishment of the γ-H2AX assay in -S₉ group (24 hours of long treatment) to establish a flow cytometry genotoxicity assay based on γ-H2AX biomarker. Two kinds of modified gold nanoparticles (sample 1 modified with cetyltrimethylammonium bromide and sample 2 modified with polyethylene glycol) were selected as experimental materials，which was diluted into three concentrations based on cytotoxicity test. The treated TK6 cells were then harvested and labeled with anti-H2AX fluorescent antibody，and Flow cytometry was used to analyze the percentage of γ-H2AX，which has been shown to be positively related to the extent of the genotoxicity. RESULTS： Both groups with and without S₉ showed significant increase in the percentage of γ-H2AX cells compared with the solvent controls (P<0.05) and these increasements were dose-related (CP：R²=0.837，P=0.01；ETO：R²=0.903，P<0.01)，thus validating the reliability of the method. On the other hand，the two gold nanoparticles products did not show significant differences in the ratio of γ-H2AX cells (P>0.05). CONCLUSION： Flow cytometry was useful in accurately detecting changes of γ-H2AX in cultured TK6 cells. This assay can be an approach for indicating the genotoxicity of toxic substances.

Key words: γ-H2AX, gold nanoparticles, flow cytometry, TK6 cell

CLC Number:

R394.6

JIANG Ruiqi, DENG Sisi, WANG Ping, CHEN Runhua, CHEN Jiaqi, ZHENG Lin, XIAN Jingwen, HUANG Zeyu, GONG Xinchao, LIU Chuheng, WANG Xiaowei, QIN Meirong. Genetic toxicity of gold nanoparticles using flow cytometry based on γ-H2AX focus analysis[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2024, 36(5): 405-411.

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Genetic toxicity of gold nanoparticles using flow cytometry based on γ-H2AX focus analysis

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