Aberrant methylation of SPARC in human hepatocellular carcinoma and its clinical implication

Figure 1 Secreted protein acidic and rich in cysteine methylation and expression in SMMC-7721 cell line. SPARC: Secreted protein acidic and rich in cysteine; Aza: 5-aza-2’-deoxycytidine; GAPDH: Glyceraldehyde 3-phosphate dehydrogenase; M: Methylation; U: Unmethylation; L: 50 bp ladder; Scale bar: 50 μm.

Figure 2 Bisulfite sequencing of secreted protein acidic and rich in cysteine in SMMC-7721 cell line. SPARC: Secreted protein acidic and rich in cysteine; TSS: Transcription start site; BS: Bisulfite sequencing; M: Methylation; U: Unmethylation; 5-Aza-CdR: 5-aza-2’-deoxycytidine; 1-18: CpG sites; - 119 to - 95, + 260 to + 282, + 52 to + 71, + 142 to + 163, + 36 to + 58, + 143 to + 167: Polymerase chain reaction primers position; Black dots: Methylation; Blank rings: Unmethylation.

Figure 3 Representative results of methylation specific polymerase chain reaction analysis and sequencing in tissues. L: 50 bp ladder; PC: Positive control; NC: Negative control; BC: Blank control; M: Methylation; U: Unmethylation; T: Hepatocellular carcinoma tissue; NT: Nontumorous tissue.

Figure 4 Expression of secreted protein acidic and rich in cysteine mRNA in hepatocellular carcinoma. Horizontal lines represent the median, and range indicates a 25%-75% quartile. SPARC: Secreted protein acidic and rich in cysteine.

Figure 5 Immunohistochemical analysis of secreted protein acidic and rich in cysteine expression. A: A tumor with positive staining; B: A tumor with negative result, but stromal tissues with positive signal (arrows); C: Nontumorous tissues with positive staining; D: Nontumorous tissues with negative staining; Scale bar: 50 μm.

Figure 6 Disease free (A) and overall (B) survival analysis of patients with different secreted protein acidic and rich in cysteine methylation status.