Down-regulation of poly (ADP-ribose) polymerase 1 leads to change of hydroquinone cytotoxicity in TK6 cells.
Abstract
Hydroquinone (HQ), one of the most important metabolites derived from benzene, is known to be associated with acute myelogenous leukemia risk; however, its carcinogenic mechanism remains unclear. In a previous study, we found that low-level of benzene exposure down-regulated the expression of poly(ADP-ribose)polymerase 1 (PARP1). Here, we employed RNA interference to knock down PARP1 expression in TK6 cells and explored the potential role of PARP1 in HQ-induced cytotoxicity. The results showed that stable PARP1-knockdown cells were successfully constructed and more than 80% inhibition of PARP1 expression was confirmed. We found that HQ treatment of TK6 cells decreased cell viability, increased cell apoptosis, and caspase3/7 activity. Knockdown of PARP1 in HQ-treated TK6 cells prevented caspase3 activation, and increased apoptosis than that in the wild-type TK6 cells, with fully functional PARP1. The results also showed that down-regulation of PARP1 led to a decrease in cell proliferation and an enhanced susceptibility to HQ-induced cytotoxicity with concentration less than 40 μM than that in normal TK6 cells. Moreover, PARP1-knockdown TK6 cells treated with HQ displayed an increased level of DNA double-strand breaks as measured by olive tail moment. No evidence was obtained of an effect of PARP1 depletion on H2AX phosphorylation induction. Under the experimental conditions, PARP1 has a role in HQ-induced DNA damage repair rather than in long-term chromatin modifications signaled by phosphorylated H2AX.