Effects of miR-7-5p on TK6 cell damage induced by hydroquinone
10.11763/j.issn.2095-2619.2017.03.005
- Author:
Hui YANG
1
;
Yuting CHEN
1
;
Hairong LIANG
1
;
Hao LUO
1
;
Yuting GAO
1
;
Lu ZHAI
1
;
Huanwen TANG
1
Author Information
1. Department of Labor Hygiene and Environmental Hygiene,School of Public Health,Guangdong Medical University,Dongguan Key Laboratory of Environmental Medicine Dongguan,Guangdong 523808,China
- Publication Type:Journal Article
- Keywords:
Hydroquinone;
Human lymphoblastoid cells;
miR-7-5p;
Cellular damage
- From:
China Occupational Medicine
2017;44(03):270-280
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE: To investigate the mechanism of miR-7-5p in TK6 cell damage induced by hydroquinone( HQ) by constructing stable miR-7-5p over-expressing human lymphoblastoid TK6 cell line using lentivirus. METHODS: i) The miR-7-5p over-expression lentivirus vectors were constructed,and then infected to TK6 cells. The miR-7-5p overexpression stable TK6 cell line( TK6-miR-7-5p cells) and negative control cells( TK6-NC cells) were selected with puromycin. The infection efficiency was confirmed by real-time fluorescent quantitative polymerase chain reaction assay.ii) TK6,TK6-NC and TK6-miR-7-5p cells were treated with HQ at final concentrations of 0 and 40 μmol/L for 48 hours.Cell viability was determined by CCK-8 assay. The early apoptosis rate of cells was detected by flow cytometry. The relative expression of poly( ADP-ribose) polymerase-1( PARP-1) and breast cancer susceptibility gene 1( BRCA1) proteins in 3 kinds of cells treated with HQ at the final concentration of 40 μmol/L was detected by Western blotting. RESULTS: i) The TK6 cell line with stable expression of miR-7-5p were successfully screened. Compared with normal TK6 cells,the relative expression of miR-7-5p in TK6-miR-7-5p cells increased by about 17 times( P < 0. 01) with no significant changes in cell morphology. ii) After treatment with 40 μmol/L HQ,the survival rate of TK6-miR-7-5p cells decreased compared with normal TK6 cells and TK6-NC cells( P < 0. 01),early apoptosis rate increased( P < 0. 01),the relative expression of PARP-1 and BRCA1 protein was decreased( P < 0. 05). CONCLUSION: MiR-7-5p may lead to the increase of early apoptosis in TK6 cells induced by HQ through inhibiting the DNA damage repair capacity related to PARP-1 and BRCA1.
