RESUMEN
Purpose The mechanism of methylation of HPV CpG sites in the occurrence and prognosis of cervical carcinogenesis remains unclear. We investigated the effects of demethylation of the CpG sites of E2 and E6, essential genes of HPV16 integration, on cervical cancer cell expression, integration, and proliferation. Materials and Methods HPV16-positive (Caski) cells were treated with different concentrations of the demethylation compound 5-aza-dc (0, 5, 10, 20 μmol/l) in vitro. After the intervention, the methylation statuses of HPV16 E2 and E6 were detected by TBS, the expression levels of E2 and E6 mRNA and protein were detected by real-time PCR and western blot, cell proliferation activity was detected by CCK8, and cell cycle and apoptosis were determined by FCM. GraphPad Prism version 8.4.2 and R version 4.2.3 were used for relevant data analyses. Results The methylation levels of HPV16 E2 and E6 CpG sites decreased gradually with increasing 5-aza-dc intervention concentrations. With decreasing E2 and E6 methylation rates, E2 expression increased, the E2/E6 ratio increased, E6 expression decreased, and the growth inhibition rate of Caski cells increased. E2 and E6 expression were negatively and positively correlated with their degrees of methylation respectively, while the E2/E6 mRNA to protein ratio was negatively correlated with the methylation degrees of E2 and E6. Conclusion Demethylation can be used as a prospective treatment to affect HPV expression and persistent infection, providing a new theoretical basis for the clinical treatment of viral infections (AU)
Asunto(s)
Humanos , Femenino , Papillomavirus Humano 16 , Proteínas Oncogénicas Virales/genética , Proteínas Oncogénicas Virales/metabolismo , Infecciones por Papillomavirus , Neoplasias del Cuello Uterino/metabolismo , Neoplasias del Cuello Uterino/virología , Proliferación Celular , Metilación de ADN , Genes Esenciales , ARN Mensajero/genética , ARN Mensajero/metabolismoRESUMEN
PURPOSE: The mechanism of methylation of HPV CpG sites in the occurrence and prognosis of cervical carcinogenesis remains unclear. We investigated the effects of demethylation of the CpG sites of E2 and E6, essential genes of HPV16 integration, on cervical cancer cell expression, integration, and proliferation. MATERIALS AND METHODS: HPV16-positive (Caski) cells were treated with different concentrations of the demethylation compound 5-aza-dc (0, 5, 10, 20 µmol/l) in vitro. After the intervention, the methylation statuses of HPV16 E2 and E6 were detected by TBS, the expression levels of E2 and E6 mRNA and protein were detected by real-time PCR and western blot, cell proliferation activity was detected by CCK8, and cell cycle and apoptosis were determined by FCM. GraphPad Prism version 8.4.2 and R version 4.2.3 were used for relevant data analyses. RESULTS: The methylation levels of HPV16 E2 and E6 CpG sites decreased gradually with increasing 5-aza-dc intervention concentrations. With decreasing E2 and E6 methylation rates, E2 expression increased, the E2/E6 ratio increased, E6 expression decreased, and the growth inhibition rate of Caski cells increased. E2 and E6 expression were negatively and positively correlated with their degrees of methylation respectively, while the E2/E6 mRNA to protein ratio was negatively correlated with the methylation degrees of E2 and E6. CONCLUSION: Demethylation can be used as a prospective treatment to affect HPV expression and persistent infection, providing a new theoretical basis for the clinical treatment of viral infections.
Asunto(s)
Proteínas Oncogénicas Virales , Infecciones por Papillomavirus , Neoplasias del Cuello Uterino , Femenino , Humanos , Neoplasias del Cuello Uterino/metabolismo , Papillomavirus Humano 16/genética , Papillomavirus Humano 16/metabolismo , Infecciones por Papillomavirus/genética , Genes Esenciales , Metilación de ADN , Proteínas Oncogénicas Virales/genética , Proteínas Oncogénicas Virales/metabolismo , Proliferación Celular , ARN Mensajero/genética , ARN Mensajero/metabolismoRESUMEN
Over the past decade, electronic markets based on the Internet, particularly online auctions, have become popular venues for conducting business. Previous studies often focused on the construction of the best bidding model, while few studies have tried to integrate multiple pricing strategies to predict the probability of closing an auction and the price premium. This study constructs a mediated model to examine the relationship among pricing strategies, the strength of bidding intentions, and online auction performance. The sample consists of 1,055 auctions of iPod MP3 players from eBay Web sites in Hong Kong, Singapore, Belgium, and France. Empirical results show that the pricing strategies directly influence both the probability of closing an auction and the level of price premium. The pricing strategies also indirectly influence the price premium through the mediating effect of the strength of bidding intentions.