Supplementary MaterialsSupplementary Information. irrespective of the duration of T2D as well as higher rates of PM of than non-DM; however, PM of was similar between both groups. PM of reduced its expression which inversely correlated with reduced patient survival. In conclusion, T2D is associated with poor prognosis of NBNC-HCC in which a high frequency of PM of may play a pivotal 4-epi-Chlortetracycline Hydrochloride role in its pathogenesis. is downregulated by PM, which may contribute to metastasis in prostate and pancreatic cancers as well as poor prognosis of gastric cancer20C22. In HCC, the reduced expression level of through PM is 4-epi-Chlortetracycline Hydrochloride inversely correlated with the expression of vascular endothelial growth factor (VEGF) and focal adhesion kinase (FAK), resulting in poor prognosis of HCC23. Notably, PM is also implicated not only in the pathogenesis of T2D but also in cancer development complicated with T2D24C26. We previously demonstrated that enhanced PM of in long-term T2D is associated with poor prognosis of pancreatic ductal cancer (PDC)26. Therefore, we hypothesised that T2D similarly enhances PM of and deteriorates the prognosis of NBNC-HCC. In this study, we evaluated the change in clinical outcome in NBNC-HCC confounded by T2D and the epigenetic modification of in T2D Methylation-specific polymerase chain reaction (MSP) was performed using primers designed for and to examine methylation (M) and unmethylation (U) in promoter regions in 10 cases followed by DNA sequencing. A positive M band indicates significant methylation of the CpG region of the promoter region (Supplemental Fig.?S1a). DNA sequencing of the MSP product confirmed methylation (Supplemental Fig.?S1b) of cytosine, which was not converted to thymine. All CpG motifs in the promoter region of were methylated. Based on previously confirmed accuracy and efficacy of MSP primers for in formalin-fixed, paraffin embedded (FFPE) specimens26, we found that the frequency of PM of in NBNC-HCC 4-epi-Chlortetracycline Hydrochloride was not significantly different between DM (43%) and non-DM controls (33%) (in the non-cancerous tissue was not significantly different between DM (26%) and non-DM (11%) (significantly increased in DM (77%) compared with that in non-DM (22%) in the cancerous tissue 4-epi-Chlortetracycline Hydrochloride (was significantly higher in DM (66%) than in non-DM (33%) (was significantly methylated in DM compared with that in non-DM irrespective of the occurrence of NBNC-HCC. Table 2 Promoter methylation analysis. was preserved in both non-cancerous and cancerous tissues in non-DM controls, whereas its expression significantly reduced in DM ((significantly reduced in non-DM, ?DPYSL3-PM (?), ?Vv (?). Survival of patients with NBNC-HCC complicated with T2DM or PM of were the significant risk factors for reduced survival (Table?3). Smoking habit and history of metabolic disorders other than T2D such as hypertension and dyslipidaemia were not correlated with DSS (Table?3). Diabetes treatment was not correlated with DSS (Supplemental Table?S1). Multivariate analysis further confirmed that hepatic vein invasion, tumour multiplicity (multiple), history of T2D and PM of remained the significant risk factors for reduced survival (Table?4). Table 3 Univariate analysis of clinicopathological factors and disease-specific survival after resection of NBNC hepatocellular carcinoma (log-rank test). promoter methylation: (?) vs (+)62.0 vs 43.50.009promoter methylation: (?) vs (+)54.0 vs 42.00.552 Open in a separate window DSS; Disease-specific survival, BMI; body mass index, T2D; type2 diabetes, wel; well-differentiated adenocarcinoma, mod; moderate-differentiated adenocarcinoma, por; poorly differentiated adenocarcinoma. Table 4 Multivariate analysis of clinicopathological factors and disease-specific survival after resection of NBNC hepatocellular carcinoma (Cox proportional hazards model). promoter methylation2.6561.210C6.4830.024 Open in a separate window 95% CI; confidence interval, T2D; type2 diabetes. Univariate analysis for overall survival (OS) showed that tumour multiplicity (multiple), serosal invasion, portal vein invasion, hepatic vein 4-epi-Chlortetracycline Hydrochloride Nrp1 invasion, T3-4 cancer [UICC stage (8th)], history of T2D and PM of were the significant risk factors for reduced survival (Supplemental Table?S2). However, smoking habit as well as a history of dyslipidaemia and hypertension was not correlated with OS (Supplemental Table?S2). Diabetes treatment was also not correlated with OS (Supplementary Table?S3). Multivariate analysis further confirmed that hepatic vein invasion, serosal invasion and history of T2D remained the significant risk factors for reduced survival (Supplemental Table?S4). The KaplanCMeier survival curve indicated a shortened DSS in DM.