研究表明，p16INK4A基因的失活是肿瘤发生发展过程中重要的分子事件。我们前期研究发现，氢醌可通过抑制PARP-1活性导致抑癌基因p16失活；另外，在氢醌作用下由PARP-1所催化的聚ADP-核糖化与蛋白磷酸化间也存在相互调控作用，提示PARP-1可能是调控p16INK4A基因转录的重要因子，但其在氢醌诱导细胞恶性转化过程中的调控机制尚不清楚。本项目拟通过氢醌诱导细胞恶性转化模型，利用质粒转染和免疫共沉淀等技术，系统研究p16/pRb通路在氢醌诱导细胞恶性转化过程中的作用；在此基础上，通过诱导和抑制PARP-1活性，证实PARP-1对p16/pRb信号通路的影响及其在氢醌诱导细胞恶性转化过程中的作用；同时，通过建立PARP-1缺陷和高表达细胞株，采用免疫沉淀、免疫印迹及反向阻断技术等，阐明PARP-1调控p16/pRb信号通路的机制，为氢醌致癌寻找新的治疗靶点提供理论和实验依据。

Some studies suggested that p16INK4A plays a critical role in carcinogenesis. Our previous study demonstrated that inhibition of p16 could be induced by decreased PARP-1 in cells exposed to hydroquinone. In addition, the reciprocal modulation existed in ribosylation and phosphorylation. These results showed that PARP-1 plays an important role in gene transcription of p16INK4A. However, the underlying mechanism in hydroquinone carcinogenesis is still unclear. Combined with the hydroquinone-induced malignant transformation of TK6 cells, plasmid transfection and immunoprecipitation, this study is aimed to investigate the effect of p16/pRb signaling pathway in the tumor formation induced by hydroquinone. On this foundation, we will use overexpression and inhibition method to confirm the regulation of p16/pRb signaling pathway by PARP-1and its role in the tumor formation induced by hydroquinone. Meanwhile, PARP-1 deficient TK6 human lymphoblastoid cell strain was established by RNA interference technique, and the high expression of TK6 human lymphoblastoid cell line of PARP-1 would be constructed. Through chromatin immunoprecipitation, Western blot and reverse blocking technology, we will clarify the molecular mechanism of PARP-1 regulation p16/pRb signaling pathway. We hope these results could provide therapeutic target and insight of preventing HQ-induce tumor formation.