Page 319 - Read Online

Shen et al. Evaluation of microRNAs normalization approaches

Figure 2: Comparisons of different microRNAs (miRNAs) panels identified with varied normalization strategies in distinguishing
hepatocellular carcinoma (HCC) tumor from non-tumor tissues. (A) A panel of 46 miRNAs identified using 3 endogenous controls as the
normalizer can distinguish HCC tumor from non-tumor tissues with 2 misclassifications. The other 3 panels of miRNAs identified by using
as normalizers of the mean of all miRNAs (B), miR-30c/miR-30b (C) and miR-30c/miR-126 (D) also classified HCC tumor from non-tumor
tissues with only 1 or 2 misclassifications

486, miR-199a-3p, miR-511, miR-424 and miR- 126 (selected as the most stable miRNAs by using
125b) were consistently dysregulated by using the NormFinder tool) as normalizers, including functionally
mean of miRNAs or 2 stable miRNAs as normalizers important oncogenic miR-21, miR-18a, miR-106a and
[Figure 3A], suggesting a high consistency for these miR-183 [Supplementary Figure 3A]. Using either the
normalization strategies. Using the mean of miRNAs mean of global miRNAs or miR-30c/miR-30b (selected
as the normalizer, we found 8 additional dysregulated as the most stable miRNAs by using geNorm tool) as
miRNAs (miR-221, miR-222, miR-324-5p, miR-550, normalizers, the same oncogenic miR-183, miR-18a
miR-362, miR-148b, miR-93# and miR-598) in HCC and miR-106a were identified [Supplementary Figure
tissue that were not significant when using 2 stable 3B]. These miRNAs would not have been identified
miRNAs as the normalizer [Supplementary Table 7]. if using 3 endogenous ncRNAs as the normalizer.
Seven dysregulated miRNAs (miR-183, miR-1180, These data suggest that the combination of using the
miR-18a, miR-130b#, miR-339-3p, miR-21 and miR- mean of miRNAs and 2 stable miRNAs identified by
106a) were identified in HCC tumor tissues using NormFinder as normalizers may be a good option to
either the mean of global miRNAs or miR-30c/miR- pinpoint biologically important miRNA.

310 Hepatoma Research ¦ Volume 2 ¦ November 18, 2016