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To explore the molecular mechanism underlying gastric carcinogenesis and progression by using gene expression profiling array together with bioinformatics. Lentivirus short hairpin RNA targeting STIL(ShSTIL)and scrambled sequence RNA(ShCon)were transduced into the gastric cancer cell line SGC-7901.RNA extraction,complementary DNA synthesis,construction of biotin-labelled amplified RNA probes,and hybridization with gene expression profile were consecutively performed.We collected corresponding data and analyzed differentially expressing genes(DEGs),followed by the analysis of gene ontology(GO)and Kyoto encyclopedia of genes and genomes(KEGG)enrichment,transcription factor regulating network,and protein-protein interacting networks. Compared with ShCon,a total of 417 and 87 genes were respectively down-regulated and up-regulated,respectively,in the ShSTIL group(1 or <-1).GO and KEGG enrichment analysis indicated that genes regulated by STIL were localized in cytoplasm,extracellular exosome,Golgi apparatus and various biomembranes,and were implicated in the ubiquitin-mediated proteolysis,P53 signaling pathway,and pathways regulating pluripotency of stem cells.Evaluation on genes enriched in KEGG pathways,regulation of transcription factors,and protein-protein interacting network demonstrated that IGF1R,STUB1,SKP2,and FOXO1 were localized at the centre of the network and played a key role in the development and progression of gastric cancer. Through the protein-protein interactions,STIL may activate E3 ubiquitin ligase STUB1 or SKP2,promote the proteolysis of FOXO1-a transcription factor,regulate the expression of IGF1R,and thus promote gastric carcinogenesis and progression.
Assuntos
Humanos , Biologia Computacional , Perfilação da Expressão Gênica , Ontologia Genética , Neoplasias Gástricas , Genética , TranscriptomaRESUMO
The hyperamplification in centrosomes give rise to a series of biochemical events including formation of multipolar spindle, abnormal segregation of chromosome and aneuploid, promoting the performance of chromosome instability, which is the main pathogenesis of multiple human malignancies.STIL participates in the formation of centrioles, activates CDK1/CyclinB1 complex and promotes mitotic entry.Moreover, it regulated expression of related downstream genes through Sonic Hedgehog signal pathway.Thus, STIL correlates with gastric and pancreatic carcinogenesis and progression.We aim to review the structure and function of STIL, association with malignancies and its potential mechanism.
RESUMO
Objective To investigate the expression pattern of STIL in gastric cancer and effect of silencing STIL on biological behaviors of SGC-7901 cell line.Methods The expression of STIL at mRNA and protein level were detected by qPCR and Western Blot,respectively.SiRNA against STIL was constructed and transfected into SGC-7901 cell line.The effect of Si-STIL on cell proliferation,cloning ability,cycle distribution and apoptosis was detected by MTT,colony-forming assay and flow cytometry,respectively.Results STIL was aberrantly expressed in gastric cancer compared with that in paratumor tissues.Downregulation of STIL level,inhibition of proliferation and cloning ability,increasing apoptosis and blocked cell cycle at S phase were observed after SGC-7901 cell line was treated with Si-STIL.Conclusion STIL was overexpressed in gastric cancer.STIL gene silencing effectively inhibited cell growth,transition of metaphase and promoted apoptosis of SGC-7901 cell line.