Characterization of 5-Hydroxymethylcytosine and Ten-to-Eleven Translocation Gene (TET) in Cervical Cancer

Cervical cancer remains one of leading causes of death in women worldwide. Epigenetic studies during the past decades have demonstrated that epigenetic silencing such as DNA methylation of tumor suppressor genes can serve as a mechanism of carcinogenesis including cervical cancer. In addition to the methylcytosine (5mC), 5-hydroxymethylcytosine (5hmC) was identified as a new epigenetic marker in mouse Purkinje neurons, the brain and embryonic stem cells in 2009, which is the first evidence that 5hmC exists in mammalian cells. In the same year, Ten-Eleven-translocation genes (TETs) was identified as molecules that catalyze the conversion of 5- methylcytosine (5mC) to 5hmC, a suggested intermediate form that potentially involved in demethylation. The understanding of these new epigenetic players, TETs and 5hmC, in cancer biology remain at its infancy. So far, most studies are investigated in leukemia. Up to date, there are no reports of 5hmC and TETs in gynecological cancers. The present project is to explore the role of this newly discovered 6th base, 5hmC, and TETs in cervical carcinogenesis. In the first part, we will test the involvement of 5hmc and TETs in cervical precancerous and cancer cell lines using dot blot and RT-PCR. We will test the protein expression in the full spectrum of cervical lesions using immunohistochemistry. We will also investigate the effects of TETs on the malignant phenotypes such as proliferation, migration, invasion and tumorigenicty of cervical cancer. In the second part, we will do the genome-wide mapping of 5hmC landscape using hMeDIP coupled with new generation sequencing techonolgy. We will identify the gene network of 5hmC targets and test the possibility of targeting 5hmC-loss-activated genes as a new therapeutics. In the third part, we will elucidate signaling pathways regulating TETs in cervical cancer, especially in relation to PTPRR and stemness. The present project is the first study of 5hmC/TETs in cervical cancer. We will have a great opportunity to discover new mechanistic insights and novel therapeutics. The understanding of hydroxymethyaltion and TETs will open a new window of cancer biology.