Sh p53

Human VSMCs (ATCC CRL-1999) were obtained from the American Type Culture Collection. VSMCs were cultured in DMEM (HyClone; GE Healthcare Life Sciences), supplemented with 1% penicillin (100 U/ml)/streptomycin (100 mg/ml) (Beyotime Institute of Biotechnology) and 10% FBS (Gibco; Thermo Fisher Scientific, Inc.) and maintained in a 37°C constant humidified incubator with 5% CO₂.
For simvastatin treatment, a total of 5×10⁵ VSMCs/ml were incubated with 2 µM simvastatin (Xingqiong Co., Ltd.) for 24 h following transduction for 24 h with the lentiviral vectors. H19 and p53 expression levels were reduced by transfecting cells with 2 mg shRNAs (lentiviral vectors) targeting H19 (sh-H19) and p53 (sh-p53), respectively, and control cells were transduced with 2 µg negative control vector (sh-NC); all shRNAs were purchased from Shanghai GenePharma Co., Ltd. VSMCs were transduced with sh-p53, sh-H19 or sh-NC in a pGLVH1/GFP+ Puro lentiviral vector (Shanghai GenePharma Co., Ltd.) using Polybrene (4 µg/ml; Merck KGaA).

Human CRC cell lines, LoVo, SW620, SW480, HCT-116, and HT-29, normal colorectal epithelial cell line, CCD-18Co and HEK293T cells were procured from ATCC (Manassas, VA). These cells were incubated in DMEM (25 mM D-glucose, 1 mM sodium pyruvate and 4 mM L-glutamine (Gibco, Carlsbad, CA) replenished with 10% FBS (Biological Industries USA, Inc., CT) and 1% penicillin–streptomycin (Gibco).
Lentiviral packaging LV5-GFP and pSIH1-H1-copGFP were used to transmit overexpressing and shRNA sequences, respectively. sh-SIRT1, sh-p53, sh-KPNA3, negative control (NC) shRNA (sh-NC), miR-101 mimic, and mimic NC were from GenePharma Co. Ltd. (Shanghai, China). The packaging lentivirus and target vectors were co-transfected into HEK293T cells employing Lipofectamine 2000. Following incubation for 48 h, supernatants were collected to detect viral titer, followed by infection of CRC cells.