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Original Research

Hypermethylation of secreted frizzled-related proteins predicts poor prognosis in non-M3 acute myeloid leukemia

, , , , , , , , & show all
Pages 3635-3644 | Published online: 20 Jul 2017

Figures & data

Table 1 Correlations of hypermethylation of SFRPs gene promoter with clinical features in non-M3 AML patients

Table 2 Primer sequences of SFRPs for RQ-PCR and RQ-MSP

Figure 1 Electrophoresis results of RQ-MSP products in leukemic cell lines.

Abbreviations: RQ-MSP, real-time quantitative methylation-specific polymerase chain reaction; M, methylation; U, unmethylation.
Figure 1 Electrophoresis results of RQ-MSP products in leukemic cell lines.

Figure 2 Electrophoresis results of RQ-MSP products of SFRPs in AML patients.

Notes: 1: GeneRuler™ 100-bp DNA ladder; 2 and 3: two controls; 4–11: samples of non-M3 AML patients (four patients with high SFRP1/SFRP2 expression and the other four with low SFRP1/SFRP2 expression); 12: positive control; 13: negative control.
Abbreviations: RQ-MSP, real-time quantitative methylation-specific polymerase chain reaction; SFRPs, secreted frizzled-related proteins; AML, acute myeloid leukemia; M, methylation; U, unmethylation.
Figure 2 Electrophoresis results of RQ-MSP products of SFRPs in AML patients.

Figure 3 Overall survival of non-M3 AML patients: (A) all patients; (B) cytogenetically normal patients.

Abbreviations: AML, acute myeloid leukemia; SFRPs, secreted frizzled-related proteins.
Figure 3 Overall survival of non-M3 AML patients: (A) all patients; (B) cytogenetically normal patients.

Figure 4 HL60 with 5-aza-dC treatment.

Notes: Relative changes in (A) SFRP1 and (B) SFRP2 expression levels and (C) SFRP1 and (D) SFRP2 methylation levels before and after treatment of HL60 cell line with the demethylating agent 5-aza-dC for 72 h at 0 μM, 0.1 μM, 1 μM, 10 μM, and 50 μM concentrations. Expression levels of SFRP1 and SFRP2 were determined by RQ-PCR and normalized to the housekeeping gene ABL. Methylation levels of SFRP1 and SFRP2 were determined by RQ-MSP and normalized to the housekeeping gene ALU. Error bars indicate standard deviation.
Abbreviations: 5-aza-dC, 5-aza-2′-deoxycytidine; RQ-PCR, real-time quantitative polymerase chain reaction; RQ-MSP, real-time quantitative methylation-specific polymerase chain reaction.
Figure 4 HL60 with 5-aza-dC treatment.

Figure 5 Correlation between SFRP1 hypermethylation and expression level in non-M3 AML patients and HL60 cell line (since the value was lg function conversion, methylation and expression level at zero was excluded).

Abbreviation: AML, acute myeloid leukemia.
Figure 5 Correlation between SFRP1 hypermethylation and expression level in non-M3 AML patients and HL60 cell line (since the value was lg function conversion, methylation and expression level at zero was excluded).