Anti mir 26a

Synthetic, chemically modified short single- or double-stranded RNA oligonucleotides: miR-26a mimics, miR-26a control (Ctrl), anti-miR-26a, anti-miR-26a Ctrl, p68 siRNA (sip68), p53 siRNA (sip53), and EZH2 siRNA (siEZH2) were synthesized from Shanghai GenePharma Co., Ltd. Commercial miR-26a expression Lentivirus and counterpart control vector were purchased from Shanghai GenePharma Co., Ltd. Commercial p68 expression plasmid and sh.p53 plasmid carrying small hairpin RNA targeting p53 were purchased from Shanghai Gene-Chem Co., Ltd. Commercial mutp53 expression plasmids and wild-type p53 expression plasmid were constructed and provided by GuangZhou Biosicen Biotechnology CO., Ltd. All oligonucleotide sequences are listed in supplementary table 1.

Antibody against ERα was from Santa Cruz (Santa Cruz, CA, USA), antibody against α-tubulin from Sigma (St Louis, MO, USA). All other reagents were from Sigma. Anti-miR-26a and nonsense of anti-miR were from GenePharma (Shanghai, PR China). Cel-miR-39 mimics were from IBS (Shanghai, PR China).

Cal27, SCC4 and SCC9 cells as well as 293T cells were obtained from ATCC (Manassas, VA, USA). UM1 cells were purchased from Japanese Collection of Research Bioresources (JCRB) cell bank (Osaka, Japan), and normal human oral keratinocyte (NHOK) was supplied by ScienCell Research Laboratories (San Diego, CA, USA). Above cells were cultured in Dulbecco’s Modified Eagle Medium (DMEM, PAN Biotech, Aidenbach, Germany) added with fetal bovine serum (FBS; 10%, HyClone, Logan, UT, USA) at 37 °C with 5% CO₂.
Mimic of miR-26a (miR-26a) or miR-26b (miR-26b) and the mimic negative control (miR-NC), inhibitors of miR-26a (anti-miR-26a) or miR-26b (anti-miR-26b) and the inhibitor negative control (anti-miR-NC) and pcDNA-PAK1 (PAK1) were acquired from GenePharma Co., ltd. (Shanghai, China). And the pcDNA empty vector (pcDNA) was purchased from Thermo Fisher Scientific (Waltham, MA, USA). SCC4 and Cal27 cells were sowed into 6-well plates (about 2 × 10⁶ cells/well) and transfected with aforementioned nucleotides or plasmids when confluence reached 70–80% using Lipofectamine 2000 (Life Technologies Corporation, Carlsbad, CA, USA) referring to manufacturer’s recommendations.