Mir 195 mimic

For overexpression or knockdown of miR-195, miR-195 mimic (5′-UAG CAG CAC AGA AAU AUU GGC-3′), miR-195 mimic NC (5′-UUC UCC GAA CGU GUC ACG UTT-3′), miR-195 inhibitor (5′-GCC AAU AUU UCU GUG CUG CUA-3′) and inhibitor-NC (5′-CAA UAU UUC UGU GCU GCU AUU-3′ all were from Shanghai GenePharma Co., Ltd. Transfection into OVCAR-3 cells were performed using Lipofectamine^® 2000 Transfection reagent (Invitrogen; Thermo Fisher Scientific, Inc.), according to the manufacturer's instructions.
To generate OVCAR-3 cells overexpressing CDC42 or CCND1, pcDNA 3.1-CDC42 and pcDNA 3.1-CCND1 (Generay Biotech Co., Ltd.) were cloned into a pcDNA 3.1 vector (Promega Corporation) and transfected into OVCAR-3 cells at 10 nM concentration. Lipofectamine^® 2000 Transfection reagent (Invitrogen; Thermo Fisher Scientific, Inc.) was used to mediate the transfection. The empty vector was used as the control. To knockdown CDC42 or CCND1 expression in OVCAR-3 cells, HS_CDC42 small interfering (si)RNA (5′-CAT CAG ATT TTG AAA ATA TTT AA-3′), HS_CCND1 siRNA (5′-GCC ACA GAT GTG AAG TTC A-3′) and NC siRNA (5′-AAT TCT CCG AAC GTG TCA CGT-3′), provided by GeneCopoeia, Inc., were transfected into OVCAR-3 cells using Lipofectamine RNAiMAX (Invitrogen; Thermo Fisher Scientific, Inc.) at 50 nM, according to the manufacturer's instructions.

miR-195 mimic (5′-UAG CAG CAC AGA AAU GGC-3′), antagomir (5′-GCC AAU AUU UCU GUG CUG CUA-3′) and scrambled negative control (NC) miRNAs (NC mimic, 5′-UUC UCC GAA CGU GUC ACG UTT-3′ and NC antagomir, 5′-CAG UAC UUU UGU GUA GUA CAA-3′) were purchased from GenePharma Co., Ltd. Cells were seeded into 96-well plates and cultured to 70-80% confluence. The cells were then transfected with miRNA mimic (100 nM) or antagomir (200 nM) using Lipofectamine^® 2000 (Invitrogen; Thermo Fisher Scientific, Inc.), according to the manufacturer's protocol. A total of 48 h after transfection, the cells were harvested.