Cell grouping and transfection

PC cells were trypsinized and inoculated into 24-well plates. After removing culture medium, the cells were classified into the following groups: i. miR-373 negative control (NC) group (cells were treated with NC for miR-373 mimics), ii. miR-373 mimics group (cells were treated with miR-373 mimics to elevate expression of miR-373), iii. shNC group (cells were treated with NC for shRNA as negative control), iv. shSIRT1 group (cells were treated with shRNA-SIRT1 to knockdown SIRT1), v. miR-373 mimics+pcDNA3.1-NC (cells were treated with miR-373 mimics followed by the pcDNA3.1 empty vector), and vi. miR-373 mimics+pcDNA3.1-SIRT1 (cells were treated with miR-373 mimics followed by pcDNA3.1- SIRT1 vector). The cells were transfected for 48 hours as above indicated, according to the guidelines for the Lipofectamine™ 2000 (Invitrogen, USA). miR-373 NC, miR-373 mimics, shNC, shSIRT1, pcDNA3.1-NC and pcDNA3.1-SIRT1 were all purchased from GenePharma (Shanghai, China).