Identification of candidate miRNAs targeting lncRNA PCAT-1 in HCC.
A and B, miR-215 inhibits PCAT-1 expression. 20 nmol/liter mimics of miR-15a, miR-16, miR-22, miR-93a, miR-105, miR-106a, miR-129, miR-138, miR-181a, miR-192, miR-205, miR-215, miR-291a, miR-294, miR-295, miR-302, miR-372, miR-373, miR-383, and miR-428, siRNA duplexes (siPCAT-1-1, siPCAT-1-2) or NC RNA were transfected into SMMC7721 and Huh7 cells. lncRNA PCAT-1 expression was detected at 48 h after transfection (left, black columns). The transfection efficiency of different miRNAs was confirmed by qRT-PCR (right, gray columns). C and D, inhibition of miR-215 up-regulates PCAT-1 expression. 20 nmol/liter of miR-215 inhibitors and NC RNA were transfected into SMMC7721 and Huh7 HCC cells. All data of PCAT-1 expression were normalized to β-actin mRNA expression levels. All miRNA expression data were normalized to U6 small RNA expression. E, schematic constructions of pGL3-PCAT-1 and pGL3-Mut215. F, pGL3-PCAT-1 and pGL3-Mut215 were co-transfected into SMMC7721 and Huh7 cells with miR-215 mimics or NC RNA. Luciferase activity was detected at 48 h after transfection and normalized relative to the Renilla luciferase expression. Inhibition effects of miR-215 mimics on pGL3-PCAT-1 or pGL3-Mut215 are shown. All results of the mean of triplicate assays with standard deviation are presented. *, p < 0.05; **, p < 0.01.