The literature search revealed a potential association between miRNAs (miR-21, -155, -196a, -196b, and -210) and pancreatic cancer or high-grade PanIN lesions , , , , ,  and ; thus, these miRNAs were evaluated. Although all five miRNAs could be
detected in the serum of the analyzed KPC mice, miR-21, -155, and -210 did not discriminate between controls, PanINs, and PC (data not shown). miR-21 levels were already increased in mice with low-grade PanIN1 and there was no greater than a two-fold increase in expression levels of miR-155 and miR-210 in the KPC mice with PC as compared to controls (data not shown). Thus, these miRNAs were excluded from further analysis. Using miR-24 as a reference and wild-type mice (n = 10) as control, we were able to consistently measure significantly increased levels of miR-196a and -196b in the serum of mice with multifocal Omipalisib molecular weight Dabrafenib concentration PanIN2/3 lesions (n = 10) and mice with invasive PC (n = 8) ( Figure 1 and Table 1). The levels of miR-196a were similar between control mice (n = 10) and KPC mice with PanIN1 lesions (n = 10) or endocrine tumors (n = 4). In contrast, mice with PanIN2/3 lesions had a median fold change of 2.7 above control/PanIN1 and mice with PC revealed a median fold change of 3.0 compared to controls and mice with PanIN1 lesions, which were both statistically significant (P = .03
and P < .01, Table 1). miR-196a had a sensitivity and a specificity of 0.9 and 0.78 for the discrimination between normal and PanIN2/3 and 0.9 and 1 for the discrimination between normal and PC, respectively. The levels of miR-196b were also similar between control mice (n = 10) and KPC mice with PanIN1 lesions (n = 10) or endocrine Palbociclib tumors (n = 4). The mice with multifocal PanIN2/3 lesions (n = 10) and invasive carcinoma (n = 8) had a median fold change in the serum levels of miR-196b of 4.2-fold and 3.6-fold compared to normal controls and mice with PanIN1 lesions ( Figure 1 and Table 1). The calculated sensitivity and specificity
for miR-196b was 0.86 and 1 for the discrimination between control and PanIN2/3 lesions and 0.86 and 0.86 for the discrimination between control and invasive cancer. The combination of both miR-196a and miR-196b attained a perfect discrimination between control and PanIN2/3 with a sensitivity and a specificity of 1. Two of the 15 samples with PanIN2/3 lesions did not have elevated miR-196a levels (cycle threshold difference values: 0.022, 1.2), but both samples revealed raised miR-196b levels (cycle threshold difference values: − 2.02, − 1.2; Figure 1, D and E). For the discrimination between normal control and invasive PC, a sensitivity of 0.86 and a specificity of 1 were calculated. Since the levels of miR-196a and miR-196b are potential diagnostic serum markers for high-grade PanIN lesions and invasive PC, we next evaluated the presence of miR-196a and -196b in human blood samples.