RESUMEN
Matrine has been shown to play a role in the suppression of gastric cancer (GC) tumorigenesis. However, whether long non-coding RNA NUT family member 2A-antisense RNA 1 (NUTM2A-AS1) is involved in matrine-induced inhibition of GC remains unknown. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, cell colony formation, and terminal deoxynucleotidyl transferase dUTP nick-end labeling assays were employed to determine the proliferation, viability, and apoptosis of GC cells, respectively. The Cancer Genome Atlas database suggested an association between NUTM2A-AS1 and GC. The reverse transcription-quantitative polymerase chain reaction was used to quantify relative levels of NUTM2A-AS1, miR-613, and vascular endothelial growth factor A (VEGFA). Reactive oxygen species generation, glutathione content, and superoxide dismutase activity were determined by corresponding reagents or assay kits. NUTM2A-AS1 knockdown led to attenuated cell viability and proliferation, as well as to enhanced apoptosis of N87 and AGS cells treated with matrine. These changes were prevented by an inhibitor of microRNA (miR)-613. Importantly, NUTM2A-AS1 expression was positively associated with tumor progression in patients with GC. NUTM2A-AS1 and miR-613 regulated the generation of reactive oxygen species, the content of glutathione, and the activity of superoxide dismutase. VEGFA served as an important effector for the NUTM2A-AS1/miR-613-regulated resistance of GC cells to matrine. These results reveal a novel mechanism of matrine resistance in GC.
Asunto(s)
MicroARNs , ARN Largo no Codificante , Neoplasias Gástricas , Alcaloides , Línea Celular Tumoral , Proliferación Celular/genética , Familia , Regulación Neoplásica de la Expresión Génica , Glutatión/metabolismo , Humanos , MicroARNs/genética , MicroARNs/metabolismo , Estrés Oxidativo , Quinolizinas , ARN sin Sentido/genética , ARN Largo no Codificante/genética , ARN Largo no Codificante/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Neoplasias Gástricas/tratamiento farmacológico , Neoplasias Gástricas/genética , Neoplasias Gástricas/metabolismo , Superóxido Dismutasa/metabolismo , Factor A de Crecimiento Endotelial Vascular/genética , Factor A de Crecimiento Endotelial Vascular/metabolismo , MatrinasRESUMEN
PURPOSE: Human TIP30 was initially identified as a candidate metastasis suppressor gene whose expression was down-regulated in human liver, lung, breast, and prostate cancers, and recently the role of this gene was examined in colorectal cancer. The aim of this study was to determine the level of TIP30 expression in colorectal carcinoma (CRC). RESULTS: TIP30 protein levels were lower in colorectal carcinomas compared to normal tissue from the control group (P < 0.001). The frequencies of hypermethylation of TIP30 in tumor were 36%, while there was no aberrant methylation in paired adjacent non-tumor tissue. A statistically significant inverse association was found between TIP30 methylation status and expression of the TIP30 protein in tumor tissues (P = 0.006). Somatic missense mutations in the TIP30 gene were identified in human CRC tissue specimens. CONCLUSIONS: Our results demonstrate that promoter methylation is involved in the decreased expression of TIP30 tumor suppressor gene in human colorectal carcinoma.