Sh flvcr1 as1 1 2

SKBR3 and MCF7 cells were transfected with specific shRNAs against FLVCR1 (sh-FLVCR1-AS1#1#2), MYC (sh-MYC) and negative control (shCtrl), as well as pcDNA3.1/CTNNB1, pcDNA3.1/MYC and the empty pcDNA3.1 vector (all purchased from GenePharma, Shanghai, China), separately. The miR-381-3p mimics, miR-381-3p inhibitor, NC mimics and NC inhibitor were simultaneously constructed by GenePharma. Transfection was conducted for 48 h in light of the protocol of Lipofectamine2000 (Invitrogen)

Human healthy cervical cells (HUCEC) and human CC cells (HeLa, Caski, C-33A, AV3) were applied (all American Type Culture Library, Rockville, MD, USA). Culture of cells was in Roswell Park Memorial Institute (RPMI) 1640 (Gibco, NY, USA) medium covering 10% fetal bovine serum (FBS) (Gibco, NY, USA) and 1% penicillin/streptomycin (Gibco, NY, USA). Selection of Hela cells is as the auxiliary research object owing to its extremely elevated difference [2 ].
Division of HeLa cells was into nine groups: The miR/sh-negative control (NC), the sh-FLVCR1-AS1#1/#2, the miR-23a-5p, the sh-FLVCR1-AS1#1 + in-NC/in-miR-23a-5p and the miR-23a-5p + Oe-NC/- Solute carrier family 7 member 11 (SLC7A11). The sh-FLVCR1-AS1#1#2 and its sh-NC with SLC7A11 and Oe-NC were employed (all Shanghai GenePharma, China). Construction of miR-23a-5p mimic/inhibitor and NC mimic/inhibitor was performed from GenePharma. Transfection of HeLa cells was via all oligonucleotides (50 nM) and vectors (4.0 μg) adopting Lipofectamine 3000 (Invitrogen) [14 (link)].