Supplementary Material for: MicroRNAs for the Prediction of Early Response to Sorafenib Treatment in Human Hepatocellular Carcinoma

Background: Several studies suggest the role of
circulating microRNAs (miRNAs) as biomarkers of hepatocellular carcinoma
(HCC). However, the serum miRNA profile associated with the response to
sorafenib remains to be elucidated. The aim of this study was to
clarify the specific miRNAs in serum that could predict the early
response of HCC to sorafenib treatment. Summary: Analyzing
the sera from 16 HCC patients, we selected five miRNAs that showed
differences in serum levels between patients with and without tumor
responses among 179 known secretory miRNAs by using locked nucleic acid
probe-based quantitative PCR. Through further analysis using a
validation cohort that included 53 HCC patients who underwent sorafenib
treatment and 8 healthy control subjects, we found that miR-181a-5p and
miR-339-5p showed significant differences in serum levels among patients
with partial response (PR), stable disease (SD), and progressive
disease (PD), where PR patients showed the highest and PD the lowest
levels. We also analyzed the factors associated with disease control
(DC; PR or SD) 3 months after the initiation of sorafenib treatment;
patients with DC showed a significantly higher level of serum
miR-181a-5p than non-DC patients or healthy control subjects (p = 0.0349
and 0.0180 for DC vs. non-DC and control vs. non-DC by Tukey-Kramer
test, respectively). We further conducted multivariate analysis among
HCC patients with Barcelona Clinic Liver Cancer stage C using
extrahepatic metastasis, serum decarboxyprothrombin, and miR-181a-5p
levels as covariables; serum miR-181a-5p was the only independent factor
for achieving DC (p = 0.0092, odds ratio 0.139, and 95% confidence
interval 0.011-0.658). In addition, miR-181a-5p level was also the only
independent factor affecting overall survival (p = 0.0194, hazard ratio
0.267, and 95% confidence interval 0.070-0.818). Key Messages: A high serum level of miR-181a-5p before treatment is associated with DC after the initiation of sorafenib.