Quantitative Real-Time PCR (qRT-PCR)

Total RNA of Hep3B and Huh-7 cells was extracted by RNA extraction Kit (TransGen Biotech, Beijing, China). TransScript miRNA First-Strand cDNA Synthesis Kit (TransGen Biotech) was utilized for the synthesis of qRT-PCR templates. TransStart Top Green qPCR SuperMix (TransGen) was used to perform qRT-PCR. The primers for miR-21-5p and internal control were purchased from Sangon Biotech (Sangon, Shanghai, China). All data were calculated by 2^−ΔΔCt method. The sequences of primers for miR-21-5p and U6 small RNA were listed as follows: U6: Forward: 5ʹ-GCGCGTCGTGAAGCGTTC-3ʹ, Reverse: 5ʹ-GTGCAGGGTCCGAGGT-3ʹ; miR-21-5p: Forward: 5ʹ-TGCGCTAGCTTATCAGACTGAT-3ʹ, Reverse: 5ʹ-CCAGTGCAGGGTCCGAGGTATT-3ʹ.