FIGURE 5.

Verification of bioinformatics prediction of miRNAs in HCC and correlation of reduction of miR‐378a‐3p with poor prognosis in HCC patients. (A) RT‐qPCR analysis of miRNAs in HCC clinical tissue specimens. The expression levels of miR‐25‐3p, miR‐101‐3p, miR‐221‐3p, miR‐378a‐3p, miR‐381‐3p, and miR‐490‐3p in 28 pairs of human HCC tumor tissues (Tumor) and adjacent nontumor liver tissues (Normal) were normalized to U6 internal control and expressed as ∆CT = (mean CT _U6 – mean CT _miRNA). *p < 0.05 or **p < 0.01 compared with Normal, Student's t test, n = 6 replicates per sample. (B) RT‐qPCR analysis of miRNAs in HCC cell lines. The expression levels of miR‐25‐3p, miR‐101‐3p, miR‐221‐3p, miR‐378a‐3p, miR‐381‐3p, and miR‐490‐3p in a panel of human HCC cell lines including HepG2, HuH7, SMMC‐7721, Li‐7, PLC/PRF5, SK‐Hep‐1, MHCC97L, and MHCC97H were normalized to U6 internal control and calculated by a 2^{−∆∆ CT} method, with 1 being the value for L‐02 normal live cell control. *p < 0.05 or **p < 0.01 compared with L‐02, Student's t test, n = 6 replicates per sample. (C) ISH analysis of miR‐378a‐3p in HCC TMA. The representative pictures of ISH (Case 55 T, –; and Case 82 T, ++) were shown. (D) Summary of miR‐378a‐3p ISH scoring. **p < 0.01 compared with Normal, Mann–Whitney U test, n = 90 cases. (E) The percentage of high and low miR‐378a‐3p expression in Tumor or Normal in HCC TMA. **p < 0.01 compared with Normal, Pearson's χ ² test, n = 90 cases. (F) The relationship between expression level of miR‐378a‐3p and prognosis of HCC patients. *p < 0.05 compared with high miR‐378a‐3p expression, Log‐rank test