RESUMEN
OBJECTIVE: The aim of this study was to elucidate the role of FOXC2-AS1 in promoting the proliferative ability and inhibiting apoptosis of melanoma by silencing p15, thereafter regulating the progression of melanoma. PATIENTS AND METHODS: FOXC2-AS1 levels in melanoma patients with or without metastasis and those with the tumor in different stages were detected by quantitative real-time polymerase chain reaction (qRT-PCR). Regulatory effects of FOXC2-AS1 on viability and apoptosis in melanoma cells were assessed, and subcellular distribution of FOXC2-AS1 was analyzed. Subsequently, the interactions of FOXC2-AS1 with EZH2 and SUZ12 were explored by RNA-Binding Protein Immunoprecipitation (RNA-RIP) assay. Through chromatin immunoprecipitation (ChIP) assay, the role of FOXC2-AS1 to regulate p15 transcription by recruiting EZH2 was verified. At last, regulatory effects of FOXC2-AS1/p15 axis on viability and apoptosis in melanoma cells were investigated. RESULTS: It was found that FOXC2-AS1 was upregulated in melanoma tissues, especially those with metastasis or stage II-IV. Melanoma patients expressing high level of FOXC2-AS1 showed worse survival than those with low level. Knockdown of FOXC2-AS1 inhibited viability, and stimulated apoptosis in A375 and sk-mel-110 cells. Besides, P15 level was upregulated in melanoma cells transfected with si-FOXC2-AS1, and FOXC2-AS1 was mainly distributed in cytoplasm. RNA-RIP assay confirmed that FOXC2-AS1 was mainly enriched in anti-EZH2 and aniti-SUZ12. Knockdown of EZH2 could markedly upregulate protein level of p15 in melanoma cells. Furthermore, it was verified that FOXC2-AS1 inhibited p15 transcription via recruiting EZH2, and the knockdown of p15 could partially reverse the regulatory effects of FOXC2-AS1 on viability and apoptosis in melanoma. CONCLUSIONS: FOXC2-AS1 stimulates proliferative ability in melanoma via silencing p15.
Asunto(s)
Inhibidor p15 de las Quinasas Dependientes de la Ciclina/metabolismo , Proteína Potenciadora del Homólogo Zeste 2/metabolismo , Melanoma/metabolismo , ARN Largo no Codificante/metabolismo , Neoplasias Cutáneas/metabolismo , Proliferación Celular , Inhibidor p15 de las Quinasas Dependientes de la Ciclina/genética , Proteína Potenciadora del Homólogo Zeste 2/genética , Humanos , Melanoma/patología , ARN Largo no Codificante/genética , Neoplasias Cutáneas/patología , Células Tumorales CultivadasRESUMEN
OBJECTIVE: The aim of this study was to evaluate the diagnostic utility of glucose transporter 1 (GLUT1) mRNA expression in bronchial brushing specimens from patients with lung cancer. METHODS: GLUT1 mRNA levels were detected by reverse transcription-polymerase chain reaction (RT-PCR) in SurePath(TM) liquid-based cytology bronchial brushing specimens from patients with lung cancer (n=76) and benign lung disease (n=154). RESULTS: Compared with patients with benign disease and compared with cytology, GLUT1 mRNA was found significantly more frequently in patients with all carcinomas, squamous cell carcinomas, adenocarcinomas and small cell carcinomas, as well as central, peripheral and diffuse carcinomas (P<0.01). Minor differences were noted in GLUT1 mRNA and cytology results between histological types and tumour location but were not statistically significant. The diagnostic performance of RT-PCR analysis of GLUT1 mRNA was significantly higher than cytology in terms of sensitivity (97.4 ± 3.6% versus 65.8 ± 10.7) and negative predictive value (98.6 ± 1.9%, versus 85.6 ± 5.1%) but specificity (90.9 ± 4.5%) and positive predictive value (84.1 ± 7.6%) were lower than cytology (100%). CONCLUSIONS: Using liquid-based cytology, RT-PCR can be performed on bronchial brushing specimens to detect GLUT1 mRNA expression, and may be a useful adjunct to cytology diagnosis. It was more sensitive than cytology but its lower specificity should be taken into account.