Fig 1: Hypermethylation (A) and expression (B) of P16 in chronic pancreatitis(A) Representative image of PCR for exon 1 and exon 2 of P16 in indicated pancreatic cancer tissues.(B) DNA was extracted from chronic pancreatitis (CP) tissues and the methylation (M) of P16 promoter was analyzed. Two (CP4 and CP5) of seven patients with chronic pancreatitis harbored methylation of the P16 promoter; CP4 was completely hypermethylated and CP5 was partially hypermethylated. (U was abbreviation of un-methylation).(C and D) Real-time PCR was used to assess the mRNA level of P16 in chronic pancreatitis (CP) and normal pancreas (NP) tissues. Results represent the mean ± SD. We found a decreased expression of P16 in chronic pancreatitis tissues. ∗, p <0.05 by Student's t test.
Fig 2: Expression of Kras, P16, P53, and Gli1 in chronic pancreatitis and PanINs(A) IHC staining for P16, Kras, and P53 in normal pancreas and pancreas harboring ADM or ADM with PanINs (scale bar, 25 μm). Blue and black arrows show the positive staining normal acinar cells and normal ductal cells, respectively. The right histogram shows the percentage of tissues cores displaying P16, Kras, and P53 staining in normal acinar cells, ADM lesion, or ADM with PanINs lesions. ∗, p <0.05 by one-way ANOVA test.(B) IHC staining for P16, Kras, and P53 in pancreas harboring low-grade or high-grade PanINs. The arrow shows the positive staining cells (scale bar, 25 μm). The right histogram shows the percentage of tissues cores displaying P16, Kras, and P53 staining in duct cells or PanINs lesions. ∗, p < 0.05 by one-way ANOVA test.(C) IHC staining for Gli1 in pancreas from chronic pancreatitis to high-grade PanINs. The image shows chronic pancreatitis, ADM lesion, low-grade PanINs, and high-grade PanINs (scale bar, 100 μm). Gli1 expression is increased in precancerous lesions compared with chronic pancreatitis. Both of the low-grade PanINs and high-grade PanINs have higher expression levels of Gli1 compared with ADM. Blank group means the group receiving control vehicle treatment. ∗, p <0.05 compared with CP group by one-way ANOVA test; #, p <0.05 compared with ADM with PanINs group by one-way ANOVA test. All results shown by bar graphs are represented as mean ± SD.
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