HCC cells were cultured to 80% confluence, washed with ice-cold PBS and subjected to RNA extraction using 1 ml of Trizol (Invitrogen, NY, USA). cDNA synthesis was carried out with the High Capacity cDNA Reverse Transcription Kit from Applied Biosystems (Carlsbad, CA, USA) and ABI TaqMan Gene Expression assays were carried out for qRT-PCR using the 7300 Real-Time PCR System (Applied Biosystems). 18S ribosomal RNA (rRNA) was measured as an internal control and the mRNA levels of target genes were normalized to the 18S rRNA level in the same sample.
