RESUMO
Long noncoding RNA (lncRNA) dysregulation is known to be taking part in majority of cancers, including osteosarcoma. In one of our previous studies, we showed that lncRNA MEG3 is being regulated by microRNA-664a (miR-664a) suppresses the migratory potential of osteosarcoma cells (U-2OS). We now report a novel lncRNA, namely, ERICD, which is linked to the transcription factor AT-rich interaction domain 3A (ARID3A) in U-2OS cells. We show that ARID3A binds to ERICD and indirectly interacts with each other via the E2F transcription factor 1 (E2F1). Furthermore, small interfering RNA (siRNA)-mediated knockdown of ERICD inhibited cell migration, formation of colonies, and proliferation in U-2OS cells. Overexpression of ARID3A inhibited cell migration, colony formation, and proliferation, whereas siRNA-mediated knockdown of ARID3A promoted cell migration, colony formation, and proliferation. Our findings indicate that ARID3A and lncRNA ERICD have plausible tumor suppressive and oncogenic functions, respectively, in osteosarcoma. Our data demonstrate the converse interaction between ARID3A and lncRNA ERICD that target DNA-binding proteins and dysregulation of their expression through E2F1 augments osteosarcoma progression. The cell rescue experiment also indicated E2F1 to be involved in the regulation of ARID3A and ERICD.
Assuntos
Proteínas de Ligação a DNA/metabolismo , Osteossarcoma/genética , RNA Longo não Codificante/genética , Fatores de Transcrição/metabolismo , Apoptose/genética , Neoplasias Ósseas/patologia , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células/genética , Proteínas de Ligação a DNA/genética , Fator de Transcrição E2F1/genética , Fator de Transcrição E2F1/metabolismo , Regulação Neoplásica da Expressão Gênica/genética , Humanos , Osteossarcoma/metabolismo , Osteossarcoma/patologia , RNA Longo não Codificante/metabolismo , Fatores de Transcrição/genéticaRESUMO
OBJECTIVE: To evaluate the performance of human leukocyte antigen (HLA)-B27 tag single nucleotide polymorphisms (SNPs) by polymerase chain reaction - restriction fragment length polymorphism (PCR-RFLP). METHODS: We genotyped three SNPs (rs116488202, rs13202464 and rs4349859). The primers were designed using Primer3 algorithm via primer-BLAST interface. PCR products were digested by using NlaIII, BmrI and TaqαI enzymes. Quality control was performed by DNA sequence analysis. RESULTS: In total, 207 patients with ankylosing spondylitis and 32 healthy controls were included in the study. The sensitivity and specificity of SNPs rs116488202 and rs4349859 in identifying HLA-B27 were identical and adequate at 0.946 and 1.000, respectively. On the other hand, the sensitivity and specificity for rs13202464 was 0.878 and 0.934, respectively. The presence of another SNP (rs141774149) in close proximity to rs116488202 complicated the analysis for RFLP and required that we sequence all the T allele carrying samples. CONCLUSION: The SNPs rs116488202 and rs4349859 may have a place in the identification of HLA-B27 in the Turkish population; however, methods other than PCR-RFLP should be considered.
