Project description:We report the application of H3K36me3 ChIP sequencing in SETD2 genotyped samples Examination of H3K36me3 in SETD2 wild-type, mutant renal cell carcinoma and SETD2 isogenic cell lines
Project description:Set domain-containing 2 (SETD2) is the most frequently mutated gene among all the histone methyltransferases (HMTs) in Clear cell renal cell carcinoma (ccRCC). Loss of function of SETD2 is significantly associated with poor prognosis in patients with ccRCC. A better understanding of the roles of SETD2 played in ccRCC can greatly improve the prognosis and quality of life of patients with kidney cancer. Clear cell renal carcinoma cell A498 were treated with si-SETD2 and si-NC, and the exosomes were extracted.
Project description:Identification of transcripts harbouring premature termination codons by NMD inhibition (GINI method) in a panel of clear cell renal cell carcinoma cell lines. Sporadic clear cell renal cell carcinoma (cRCC) is genetically characterized by the recurrent loss of chromosome 3p, with a hotspot for copy number loss in the 3p21 region. In this study, we applied a method called Gene Identification by Nonsense Mediated mRNA decay Inhibition (GINI) on a panel of 10 cRCC cell lines with 3p21 copy number loss to identify biallelic inactivated genes located at 3p21. This analysis revealed inactivation of the histone methyltransferase gene SETD2, located on 3p21.31, as a common event in cRCC cells. SETD2 is nonredundantly responsible for trimethylation of the histone mark H3K36. Consistent with this function, we observed loss or decrease of H3K36me3 in 7 out of 10 cRCC cell lines. Identification of missense mutations in 2 of 10 primary cRCC tumor samples further supported the involvement of loss of SETD2 function in the development of cRCC tumors.
Project description:In this study, SETD2 null isogenic 38E/38F clones derived from 786-O cells were generated by zinc finger nucleases, and the cellular metabolic changes of 786-O (WT) and 38E/38F isogenic cell lines (n=3 per group) were analyzed by GC-MS-based targeted metabolomics.
