Si hotair

Small interfering RNA specific for HOTAIR (si-HOTAIR) and control siRNA (si-NC) was synthesized (Ribobio, Guangzhou, China) and transfected using Lipofectamine 2000 in CaSki and HeLa cells. The sequences of siRNA were: si-HOTAIR1, 5′-AAAUCCAGAACCCUCUGACAUUUGC-3′, si-HOTAIR2, 5′-UU AAGUCUAGGAAUCAGCACGAAGC-3′ and si-HOTAIR3, 5′-CAUAUUAUAGA GUUGCU CUGUGCUG-3′.

For overexpression of HOTAIR and FUT2, HOTAIR or FUT2 cDNA was cloned into the multiple cloning site of the pcDNA3.1 vector (Invitrogen, Carlsbad, CA, USA). MiR-17-5p mimic, negative control oligonucleotides (miR-NC), miR-17-5p inhibitor, negative control oligonucleotide (NC inhibitor), small interfering RNA of HOTAIR or FUT2 (siHOTAIR, siFUT2), scramble siRNA of HOTAIR or FUT2 (siSCR) were purchased from RiboBio (Guangzhou, China). The chondrocytes were seeded into 6-well plates and transfection was performed by using Lipofectamine 3000 (Invitrogen, Carlsbad, CA, USA). After 48 h of transfection, chondrocytes were stimulated with IL-1β (10 ng/ml) for the 24 h and used for further analysis.

HOTAIR cDNAs were cloned to mammalian expression vector pcDNA3.1 (Invitrogen, USA). Small interfering RNA (siRNA) targeting HOTAIR (ie si‐HOTAIR) and controlled siRNA (si‐NC) were purchased from Ribobio (China). Gene Pharmaceutical cor. (China) designed and synthesized the miR‐17‐5p mimics, miR‐17‐5p inhibitor, and miR‐NC. According to the manufacturer's instructions (Invitrogen, USA), Lipofectamine 2000 was adopted for the transfection process.