Cmir an0001 sn

miR-345 mimic (HmiR0210-MR03; GeneCopoeia, Inc., Guangzhou, China) and control vector (CmiR0001-MR03; GeneCopoeia) were transfected into MHCC-97H cells while miR-345 inhibitor (HmiR-AN0437-AM01; GeneCopoeia) and negative control vector (CmiR-AN0001-SN; GeneCopoeia) were transfected into Hep3B cells. YAP1 overexpression plasmid and the control plasmid were transfected into Hep3B cells overexpressing miR-345 while YAP1 siRNA and scramble siRNA were transfected into MHCC-97H cells with miR-345 knockdown. Cell transfection of HCC cells were carried out in 6-well plates with Lipofectamine 2000 (Invitrogen, Waltham, MA, USA). Cells after transfection were collected for western blot analysis, qRT-PCR, would healing assay, Transwell assay and in vivo experiments.

The MEG3 sequence was synthesized based on the MEG3 sequence and then sub-cloned into the pcDNA3.1 vector (pcDNA-MEG3; Shanghai GeneChem Co., Ltd.). The empty pcDNA3.1 vector was used as a control (pcDNA-control). For MEG3 knockdown, the MEG3-shRNA plasmid (sh-MEG3 sequence, 5′-GAGAGGTTGTTTCACTGGTATCTATTGCA-3′; pGFP-C-shLenti Vector; cat. no. TL 320132C) and the scrambled shRNA plasmid (sh-control; pGFP-C-shLenti Vector; cat. no. TR30021) were purchased from OriGene. HTR-8/SVneo cells were seeded into 6-well plates (1×10⁶ cells/well) and cultured at 37°C for 24 h. The cells were subsequently transfected with 100 ng pcDNA-control, 100 ng pcDNA-MEG3, 100 ng sh-control, 100 ng sh-MEG3, 100 nM miR-345-3p inhibitor (cat. no. HmiR-AN0437-AM01; GeneCopoeia, Inc.), 100 nM inhibitor control (cat. no. CmiR-AN0001-SN; GeneCopoeia, Inc.) or 100 ng sh-MEG3 + 100 nM miR-345-3p inhibitor using Lipofectamine^® 2000 reagent (Invitrogen; Thermo Fisher Scientific, Inc.) according to the manufacturer's protocols. Transfection efficiency was assessed using reverse transcription-quantitative PCR (RT-qPCR) 48 h after transfection.