6 ± 11 8 0 709 53 6 ± 18 7 0 265 56 5 ± 11 9 0 337    Female 15 5

6 ± 11.8 0.709 53.6 ± 18.7 0.265 56.5 ± 11.9 0.337    Female 15 59.8 ± 12.1   55.5 ± 22.6   58.0 ± 13.2   Age (yrs)                  ≤ 55 19 58.0 ± 12.0 0.386 52.6 ± 19.1 0.156 55.7 ± 12.1 0.142    > 55 21 60.0 ± 11.7   56.0 ± 21.0   58.3 ± 12.6   Alcohol                  – 20 58.7 ± 12.9 0.794 46.6 ± 18.2 0.016

53.7 ± 11.2 0.154    + 20 60.0 ± 11.7   62.1 ± 19.1   60.5 ± 12.6   Smoking                  – 22 58.1 ± 13.7 0.671 47.5 ± 17.5 0.017 53.7 ± 11.9 0.067    + 18 60.2 ± 9.1   62.8 ± 19.1   61.3 ± 11.7   Tumor size (cm)                  ≤ 2 21 55.4 ± 10.5 0.087 46.1 ± 18.8 0.029 51.5 ± 10.1 0.013    > 2 19 63.1 ± 12.0   63.5 ± 17.4   63.3 ± 11.7   Differentiation learn more                  Moderate 19 59.6 ± 12.2 0.625 53.6 ± 20.4 0.799 57.1 ± 12.4 0.877    Poor 21 58.6 ± 11.6   55.0 ± 20.1   57.1 ± 12.5   Lymph node metastasis                  – 23 60.4 ± 12.4 0.307 53.7 ± 20.0 0.832 57.6 ± 12.5 0.421    + check details 17 57.2 ± 10.9   55.2 ± 20.7   56.4 ± 12.3   pTNM stage                  I+II 21 58.2 ± 12.4 0.444 51.9 ± 20.1 0.867 55.5 ± 12.6 0.543    III+IV 19 60.0 ± 11.2   57.1 ± 20.0   58.8 ± 12.0   Correlations of SPARC methylation with clinical characteristics of pancreatic cancer were determined by general linear model univariate analysis. Table 2 The standardized coefficient beta value of multiple regression

analysis Clinical characteristics Region 1 Region 2 Whole region

Gender — – — Age — – — Alcohol — 0.341 (p = 0.012) — Smoking — 0.336 (p = 0.013) — Tumor size 0.332 (p = 0.036) 0.342 (p = 0.013) 0.485 (p = 0.002) Differentiation — – — Lymph node metastasis — – — pTNM stage — – — Adjusted Pyruvate dehydrogenase R 2 0.087 0.367 0.215 Clinical characteristics of pancreatic cancer were analyzed using a stepwise multiple regression to assess their independent contribution to the methylation level, with entry and removal at the 0.05 and 0.1 significance levels, respectively. Discussion In the current study, we determined the methylation status of the SPARC gene promoter in pancreatic cancer cell lines, pancreatic cancer and corresponding adjacent normal pancreatic tissues, chronic pancreatitis tissues, and real normal pancreatic tissues. Methylation of the SPARC gene TRR gradually increased from normal, chronic pancreatitis, and the adjacent normal tissues to pancreatic cancer tissues. The methylation pattern of the SPARC gene TRR exhibited two hypermethylation wave peak regions: CpG Region 1 (CpG site 1-7) and CpG Region 2 (CpG site 8-12). CpG Region 2 was rarely methylated in real normal pancreatic tissues but CpG Region 1 was more frequently methylated. In addition, the methylation level of CpG Region 2 in the adjacent normal tissues was significantly increased compared with the real normal tissues.

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