RESUMO
Human UHRF1 (ubiquitin-like PHD and RING finger domain-containing 1) has been reported to be over-expressed in many cancers, but its role in ovarian cancer remains elusive. Here, we determined whether knockdown of UHRF1 by lentivirus-mediated shRNA could inhibit ovarian cancer cell growth. Lentivirus- mediated short hairpin RNAs (lv-shRNAs-UHRF1) were designed to trigger the gene silencing RNA interference (RNAi) pathway. The efficiency of lentivirus-mediated shRNA infection into HO-8910 and HO-8910 PM cells was determined using fluorescence microscopy to observe lentivirus-mediated GFP expressionand was confirmed to be over 80 percent. UHRF1 expression in infected HO-8910 and HO-8910 PM was evaluated by real-time PCR and Western blot analysis. The Cell Counting Kit-8 (CCK-8) assay was used to measure cell viability; flow cytometry and Hoechst 33342 assay was applied to measure cell cycle arrest and apoptosis. Cell invasion was assessed using transwell chambers. Our results demonstrated that the loss of UHRF1 promoted HO-8910 and HO-8910 PM cell apoptosis, while inhibiting cell proliferation. In addition, UHRF1 knockdown significantly inhibited the invasion of human ovarian cancer cells. In the present study, we also showed that depleting HO-8910 cells of UHRF1 caused activation of the DNA damage response pathway, with the cell cycle arrested in G2/M-phase. The DNA damage response in cells depleted of UHRF1 was illustrated by phosphorylation of CHK (checkpoint kinase) 2 on Thr68, phosphorylation of CDC25 (cell division control 25) on Ser 216 and phosphorylation of CDK1 (cyclin-dependent kinase 1) on Tyr 15.
Assuntos
Biomarcadores Tumorais/genética , Proteínas Estimuladoras de Ligação a CCAAT/antagonistas & inibidores , Movimento Celular , Proliferação de Células , Lentivirus/genética , Neoplasias Ovarianas/prevenção & controle , RNA Interferente Pequeno/genética , Apoptose , Western Blotting , Proteínas Estimuladoras de Ligação a CCAAT/genética , Proteínas Estimuladoras de Ligação a CCAAT/metabolismo , Ciclo Celular , Feminino , Citometria de Fluxo , Humanos , Neoplasias Ovarianas/genética , Neoplasias Ovarianas/patologia , RNA Mensageiro/genética , Reação em Cadeia da Polimerase em Tempo Real , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Células Tumorais Cultivadas , Ubiquitina-Proteína LigasesRESUMO
OBJECTIVE: To investigate the prevalence of Entamoeba gingivalis infection in college students in Tangshan, and analyze the relationship between the infection and human behaviors. METHODS: 551 students of grades 1-3 from six colleges in Tangshan received questionnairing, which covered the oral health state, teeth-brushing, xylitol gum-chewing, diet fondness, and smoking. Specimens were taken from the tooth surface of the lesion or fouling materials by using disinfected toothpicks and the smears were observed microscopically to examine Entamoeba gingivalis infection. RESULTS: The prevalence of Entamoeba gingivalis infection was 28.3% (156/551), 30.4% (55/181) and 24.6% in males and females (91/370) respectively (chi2=2.09, P>0.05). The prevalence in students with or without oral disorders was 41.2% (84/204) and 20.8% (72/347) respectively, with a significant statistical difference (chi2=26.41, P<0.01); it was 22.5% (53/236) and 32.7% (103/315) among students who cleaned their teeth regularly or irregularly (chi2=6.97, P<0.01); it was 18.3% (17/93) and 30.4% (139/458) among those usually with or without chewing xylitol gum (chi2=5.55, P<0.05). CONCLUSION: Entamoeba gingivalis infection is common in the college students in Tangshan and it has a close relation to the oral hygiene habits and the presence of oral disorders.