Effect of RICTOR expression on cell viability in rheumatoid arthritis fibroblast-like synoviocytes |
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Authors: | GUO Xin HU Ai-ling FANG Lin-kai LIU Yan PAN Yun-feng |
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Affiliation: | 1.Department of Rheumatology, 2Department of Nursing, the Third Affiliated Hospital, Sun Yat-sen University, Guangzhou 510630, China. |
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Abstract: | AIM:To investigate the effects of RICTOR expression on the viability of rheumatoid arthritis fibroblast-like synoviocytes (RA-FLS). METHODS:RA-FLS were obtained by tissue culture. Chemically synthesized double-stranded siRNAs targeting RICTOR gene were transfected into RA-FLS by cationic liposome. The nonspecific siRNA was also transfected into the negative control cells. The mRNA expression of RICTOR was detected by RT-qPCR after transfection for 24 h. Western blotting was used to evaluate the inhibitory effects of siRNAs on RICTOR expression after transfection for 48 h and 72 h. The cell viability was examined by MTT assay. RESULTS:Compared with control group, the mRNA expression of RICTOR significantly decreased by 78.36%±3.71% after the transfection of RICTOR siRNA for 24 h. The protein level of RICTOR was also obviously lower in RICTOR siRNA transfection group than that in control group after transfection for 48 h (decreased by 92.48%±6.14%) and 72 h (decreased by 98.57%±1.40%). Knock-down of RICTOR in RA-FLS for 72 h markedly decreased the cell viability. CONCLUSION:Transfection of RICTOR siRNA reduces the viability of RA-FLS, indicating that mTORC2 may be required for the survival of RA-FLS. |
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Keywords: | RICTOR protein Arthritis rheumatoid Fibroblast-like synoviocyte RNA interfering |
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