ABSTRACT
MYC overexpression is a common phenomenon in gastric carcinogenesis. In this study, we identified genes differentially expressed with a downregulated profile in gastric cancer (GC) cell lines with silenced MYC. The TTLL12, CDKN3, CDC16, PTPRA, MZT2B, UBE2T genes were validated using qRT-PCR, western blot and immunohistochemistry in tissues of 213 patients with diffuse and intestinal GC. We identified high levels of TTLL12, MZT2B, CDC16, UBE2T, associated with early and advanced stages, lymph nodes, distant metastases and risk factors such as H. pylori. Our results show that in the diffuse GC the overexpression of CDC16 and UBE2T indicate markers of poor prognosis higher than TTLL12. That is, patients with overexpression of these two genes live less than patients with overexpression of TTLL12. In the intestinal GC, patients who overexpressed CDC16 had a significantly lower survival rate than patients who overexpressed MZT2B and UBE2T, indicating in our data a worse prognostic value of CDC16 compared to the other two genes. PTPRA and CDKN3 proved to be important for assessing tumor progression in the early and advanced stages. In summary, in this study, we identified diagnostic and prognostic biomarkers of GC under the control of MYC, related to the cell cycle and the neoplastic process.
Subject(s)
Adenocarcinoma/genetics , Proto-Oncogene Proteins c-myc/genetics , Stomach Neoplasms/genetics , Adenocarcinoma/metabolism , Adenocarcinoma/mortality , Apc6 Subunit, Anaphase-Promoting Complex-Cyclosome/genetics , Apc6 Subunit, Anaphase-Promoting Complex-Cyclosome/metabolism , Cell Line, Tumor , Cyclin-Dependent Kinase Inhibitor Proteins/genetics , Cyclin-Dependent Kinase Inhibitor Proteins/metabolism , Down-Regulation , Dual-Specificity Phosphatases/genetics , Dual-Specificity Phosphatases/metabolism , Female , Gene Silencing , Humans , Male , Microtubule-Associated Proteins/genetics , Microtubule-Associated Proteins/metabolism , Middle Aged , Peptide Synthases/genetics , Peptide Synthases/metabolism , Prognosis , RNA, Small Interfering , RNA-Seq , Receptor-Like Protein Tyrosine Phosphatases, Class 4/genetics , Receptor-Like Protein Tyrosine Phosphatases, Class 4/metabolism , Stomach Neoplasms/metabolism , Stomach Neoplasms/mortality , Ubiquitin-Conjugating Enzymes/genetics , Ubiquitin-Conjugating Enzymes/metabolismABSTRACT
Cell division cycle16 (CDC16) is a core component among the eight protein subunits of the anaphase-promoting complex (APC). APC is a cyclin degradation system that governs the exit of cells from mitosis. Not much information is available for CDC16 in pig. In this study, a 2284-bp cDNA of porcine CDC16 was obtained by rapid amplification of cDNA ends (RACE). Porcine CDC16 was assigned to SSC11 q11-17, and was determined to be significantly linked with SW1452 by using somatic cell hybrid panel and radiation hybrid panel. One novel A/G SNP anchored in intron 7 of the gene was genotyped by restriction enzyme polymerase chain reaction (PCR)-restriction fragment length polymorphism-Csp6I. In five pig breeds, Shaziling, Taoyuan, Duroc, Landrace, and Yorkshire, the A allele frequency was dominant. Quantitative PCR revealed that porcine CDC16 was expressed in ten selected tissues of 25-day-old Shaziling and Yorkshire piglets, and that the mRNA expression of CDC16 in longissimus dorsi muscle of Shaziling was higher than that of Yorkshire. Expression levels of CDC16 were highest in longissimus dorsi muscle followed by that in pancreas. CDC16 protein was detected in longissimus dorsi muscle of 25-day-old Shaziling and Yorkshire piglets by immunohistochemistry with abundant protein expression index (P > 0.05). This study provides an insight into the role of porcine CDC16 in the formation of meat.
Subject(s)
Apc6 Subunit, Anaphase-Promoting Complex-Cyclosome/genetics , Chromosome Mapping/methods , Animals , Gene Expression Profiling , Gene Frequency/genetics , Genotype , Polymorphism, Restriction Fragment Length , SwineABSTRACT
Rab GTPases serve as major control elements in the coordination and definition of specific trafficking steps and intracellular compartments. Rab activity is modulated in part by GTPase-activating proteins (GAPs), and many RabGAPs share a Tre-2/Bub2/Cdc16 (TBC)-domain architecture, although the majority of TBC proteins are poorly characterized. We reconstruct the evolutionary history of the TBC family using ScrollSaw, a method for the phylogenetic analysis of pan-eukaryotic data sets, and find a sophisticated, ancient TBC complement of at least 10 members. Significantly, the TBC complement is nearly always smaller than the Rab cohort in any individual genome but also suggests Rab/TBC coevolution. Further, TBC-domain architecture has been well conserved in modern eukaryotes. The reconstruction also shows conservation of ancestral TBC subfamilies, continuing evolution of new TBCs, and frequent secondary losses. These patterns give additional insights into the sculpting of the endomembrane system.
