Si tnf α

Cells were incubated in seven groups: Negative control (NC), si-MMP14, miR-195-5p-mimics, miR-195-5p-inhibitor, si-TNF-α and si-MMP14 + miR-195-5p-inhibitor. NC, miR-195 mimics, miR-195 inhibitor, si-TNF-α and si-MMP14 were compounded by Shanghai GenePharma Co., Ltd. (Shanghai, China). Cells in the exponential phase of growth were seeded in a 6-well plate and maintained in DMEM without antibiotics. The transfection was carried out using Lipofectamine™ 2000 (Invitrogen; Thermo Fisher Scientific, Inc.) following the manufacturer's directions when the cell density reached 50-60%.
qRT-PCR TRIzol reagent (Invitrogen; Thermo Fisher Scientific, Inc.) was employed for total RNA (tRNA) isolation. PrimeScript™ RT Reagent kit (Takara Biotechnology Co., Ltd., Dalian China) was used for cDNA synthesis using 1 μg tRNA. QRT-PCR was carried out using SYBR Premix Ex Taq Master mix (Takara Biotechnology Co, Ltd.) in an Applied Biosystems 7500 Real-Time PCR system (Thermo Fisher Scientific, Inc.) in accordance with the manufacturer's directions. U6 small nuclear RNA (U6) and GAPDH were used as internal controls for miR-195-5p and MMP14 mRNA expression, respectively. The involved primers were present in Table 1. The data were analyzed using 2 -∆∆Ct method. All experiments were performed for three times.