Project description:<p>Comprehensive genomic profiling of colon adenocarcinomas has revealed multiple recurrent alterations that may inform new treatment strategies. Clinical trials of these agents are ongoing, although the mechanisms of response and resistance to these agents are not well characterized. The goal of this study is to perform comprehensive profiling of pre-treatment and post-resistance tumor and germline samples obtained from patients with colon cancer who receive these agents by carrying out whole exome sequencing and RNA sequencing, and to use these data to identify mechanisms of response and resistance.</p>
Project description:Using the highly sensitive miRCURY LNA™ microRNA array, we screened 3100 microRNAs abundant in the human colon cancer and Adjacent normal gastric mucosa tissues, and the function of differentially expressed microRNAs were analyzed by bioinformatics. The enrichment results indicated that these microRNAs perhaps participated in the occurrence and development process of colon cancer. In this study, three cases of colon cancer were used to acquire the microRNA expression profiling, and qPCR was employed to confirm the results of microRNA chip. The function of the differentially expressed microRNA were analyzed by bioinformatic methods.Finally,compared with the adjacent normal mucosa tissues,the expression of 2 microRNAs were up regulated in colon cancer. qPCR results showed an expression pattern consistent with that of the chip analysis.
Project description:N6-methyl adenosine (m6A) is one of the most important RNA modifications involved in several biological and pathological processes, including cancer. Dysregulation of m6A has been linked with tumor initiation, progression, and metastasis of several cancer types, including colon cancer. A transcriptome of colon cancer describes the dysregulated coding and non-coding RNAs but does not reveal the mechanisms like m6A modifications that determine the post-transcriptional and pre-translational regulations. Epi-transcriptome profiling of m6A in colon cancer cell lines was performed using Methylated RNA Immunoprecipitation (MeRIP) sequencing. Overall, the study illustrates the distribution of m6A across the transcriptome of various colon cancer cell lines.
Project description:In this study, paired tumors and distant normal tissues (DNTs) collected from 78 treatment-naïve colon cancer patients were subjected to tandem mass tag (TMT)-based proteomics. We found that the expression of GSK3α, but not GSK3β, was significantly correlated with the overall survival (OS) of colon cancer patients in 3 independent cohorts. To decipher the roles of GSK3α in colon cancer, we profiled the phosphorylation substrates of GSK3α. Initial in vitro and in vivo phosphoproteomics analyses uncovered 156 phosphosites from 130 proteins specifically regulated by GSK3α but not GSK3β. Notably, the levels of some phosphosites, including HSF1S303p, CANXS583p, MCM2S41p, POGZS425p, SRRM2T983p and PRPF4BS431p, were significantly correlated with the OS of colon cancer patients. Further pull-down assays identified 23 proteins, such as THRAP3, BCLAF1 and STAU1, that showed strong binding affinity to GSK3α. The tight interaction between THRAP3 and GSK3α was verified by biochemical experiments. Notably, among the 18 phosphosites of THRAP3, phosphorylation at S248, S253 and S682 is specifically mediated by GSK3α in vitro and in cell lines. Mutation of S248 to D (S248D), which mimics the effect of phosphorylation, obviously increased cancer cell migration and the binding affinity to proteins related to DNA damage repair. Collectively, this work not only discloses the specific function of GSK3α as a kinase but also suggests GSK3α as a promising therapeutic target for colon cancer.