AIM:To assess diagnostic accuracy of Ras association domain family 1A(RASSF1A)promoter methylation in body fluids(serum,plasma and whole blood)for hepatocellular carcinoma(HCC).METHODS:Relative information about study...AIM:To assess diagnostic accuracy of Ras association domain family 1A(RASSF1A)promoter methylation in body fluids(serum,plasma and whole blood)for hepatocellular carcinoma(HCC).METHODS:Relative information about study characteristics and incidence of RASSF1A methylation was collected.Quality of all included studies was evaluated by Quality Assessment of Diagnostic Accuracy Studies-2.Sensitivity and specificity were pooled using a randomeffect model,and a summary receiver operating characteristic curve was used to demonstrate the overall diagnostic performance.Positive likelihood ratio(PLR),negative likelihood ratio(NLR),and diagnostic odds ratio(DOR)with 95%CI were also calculated.Meta-regression was applied to analyze observed heterogeneity,and Deeks’test was performed to detect publication bias.RESULTS:After a systematic literature review,seven studies with a total of 302 cases of HCC and 250 cases of chronic liver diseases were included in the analysis.The pooled sensitivity and specificity were 0.70(95%CI:0.49-0.85)and 0.72(95%CI:0.54-0.85),respectively.The PLR was 2.51(95%CI:1.64-3.86),NLR was 0.41(95%CI:0.25-0.68),and DOR was 6.13(95%CI:3.17-11.84).Theχ2values of sensitivity,specificity,PLR,NLR and DOR were 59.41(P<0.001),50.50(P<0.001),17.40(P=0.010),31.24(P<0.001)and80.51(P<0.001),respectively.The area under the curve was 0.77(95%CI:0.73-0.81).Three factors were analyzed by univariate meta-regression and none was significant to interpret the observed heterogeneity(P>0.05).No significant publication bias was detected by Deeks’test(P=0.346).CONCLUSION:We showed the potential diagnostic value of RASSF1A methylation in body fluids in HCC patients and it may improve diagnostic accuracy combined with theα-fetoprotein test.展开更多
AIM: To investigate the effect of 5-Aza-2’-deoxycytidine(5-Aza-CdR),a DNA methyltransferase(DNMT) inhibitor,on the growth and survival of the Chinese retinoblastoma(RB) cell line HXO-RB44. ·METHODS: The DNA meth...AIM: To investigate the effect of 5-Aza-2’-deoxycytidine(5-Aza-CdR),a DNA methyltransferase(DNMT) inhibitor,on the growth and survival of the Chinese retinoblastoma(RB) cell line HXO-RB44. ·METHODS: The DNA methylation status of the Ras association domain family(RASSF1A) promoter in the presence of 5-Aza-CdR at different concentrations was analyzed by methylation-specific polymerase chain reaction(MSP). RASSF1A mRNA and protein levels were measured by semiquantitative RT-PCR and immunohistochemistry staining,respectively,when cells were treated with 5.0μmol/L of 5-Aza-CdR. The effect of 5.0μmol/L 5-Aza-CdR on the proliferation and viability of HXO-RB44 cells was examined using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay and flow cytometry. ·RESULTS: 5-Aza-CdR efficiently induced cell cycle arrest at G0 /G1 and apoptotic death in HXO-RB44 cells. MSP analysis showed that unmethylated RASSF1A DNA increased and methylated RASSF1A decreased in a dose-dependent manner in a range of 0.5-5.0μmol/L 5-Aza-CdR. Accordingly,RASSF1A expression was reactivated at both mRNA and protein levels. Incubation time of 5-Aza-CdR treatment also functioned as a factor for the demethylation status of RASSF1A promoter DNA,with a plateau on day four. 5-Aza-CdR at 5.0μmol/L completely demethylated the RASSF1A promoter in HXORB44 cells on day four,and as a result,RASSF1A expression increased significantly from day 4 to day 7.·CONCLUSION: 5-Aza-CdR inhibits the growth of the HXO-RB44 RB cell line and induces apoptosis by demethylating the RASSF1A gene.展开更多
基金Supported by Key Project of Chinese Ministry of Science and Technology,No.2012ZX10002007 and No.2013ZX10002001National Natural Science Foundation of China,No.81171579and No.81201287Natural Science Foundation of Shandong Province,China,No.ZR2010HM070 and No.ZR2010HQ040
文摘AIM:To assess diagnostic accuracy of Ras association domain family 1A(RASSF1A)promoter methylation in body fluids(serum,plasma and whole blood)for hepatocellular carcinoma(HCC).METHODS:Relative information about study characteristics and incidence of RASSF1A methylation was collected.Quality of all included studies was evaluated by Quality Assessment of Diagnostic Accuracy Studies-2.Sensitivity and specificity were pooled using a randomeffect model,and a summary receiver operating characteristic curve was used to demonstrate the overall diagnostic performance.Positive likelihood ratio(PLR),negative likelihood ratio(NLR),and diagnostic odds ratio(DOR)with 95%CI were also calculated.Meta-regression was applied to analyze observed heterogeneity,and Deeks’test was performed to detect publication bias.RESULTS:After a systematic literature review,seven studies with a total of 302 cases of HCC and 250 cases of chronic liver diseases were included in the analysis.The pooled sensitivity and specificity were 0.70(95%CI:0.49-0.85)and 0.72(95%CI:0.54-0.85),respectively.The PLR was 2.51(95%CI:1.64-3.86),NLR was 0.41(95%CI:0.25-0.68),and DOR was 6.13(95%CI:3.17-11.84).Theχ2values of sensitivity,specificity,PLR,NLR and DOR were 59.41(P<0.001),50.50(P<0.001),17.40(P=0.010),31.24(P<0.001)and80.51(P<0.001),respectively.The area under the curve was 0.77(95%CI:0.73-0.81).Three factors were analyzed by univariate meta-regression and none was significant to interpret the observed heterogeneity(P>0.05).No significant publication bias was detected by Deeks’test(P=0.346).CONCLUSION:We showed the potential diagnostic value of RASSF1A methylation in body fluids in HCC patients and it may improve diagnostic accuracy combined with theα-fetoprotein test.
基金Supported by the National Natural Science Foundation of China(No.3087282No.81072221)
文摘AIM: To investigate the effect of 5-Aza-2’-deoxycytidine(5-Aza-CdR),a DNA methyltransferase(DNMT) inhibitor,on the growth and survival of the Chinese retinoblastoma(RB) cell line HXO-RB44. ·METHODS: The DNA methylation status of the Ras association domain family(RASSF1A) promoter in the presence of 5-Aza-CdR at different concentrations was analyzed by methylation-specific polymerase chain reaction(MSP). RASSF1A mRNA and protein levels were measured by semiquantitative RT-PCR and immunohistochemistry staining,respectively,when cells were treated with 5.0μmol/L of 5-Aza-CdR. The effect of 5.0μmol/L 5-Aza-CdR on the proliferation and viability of HXO-RB44 cells was examined using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay and flow cytometry. ·RESULTS: 5-Aza-CdR efficiently induced cell cycle arrest at G0 /G1 and apoptotic death in HXO-RB44 cells. MSP analysis showed that unmethylated RASSF1A DNA increased and methylated RASSF1A decreased in a dose-dependent manner in a range of 0.5-5.0μmol/L 5-Aza-CdR. Accordingly,RASSF1A expression was reactivated at both mRNA and protein levels. Incubation time of 5-Aza-CdR treatment also functioned as a factor for the demethylation status of RASSF1A promoter DNA,with a plateau on day four. 5-Aza-CdR at 5.0μmol/L completely demethylated the RASSF1A promoter in HXORB44 cells on day four,and as a result,RASSF1A expression increased significantly from day 4 to day 7.·CONCLUSION: 5-Aza-CdR inhibits the growth of the HXO-RB44 RB cell line and induces apoptosis by demethylating the RASSF1A gene.
