2022, Vol. 26 ›› Issue (32): 5112-5118

Activation of chondrocyte degeneration related signaling pathways and expression of differential factors in ATDC5 mouse cells after silent information regulator 1 knockdown

Xie Xiaochen1, 2, Yang Qi3, Weng Jian1, 2, Zeng Hui1, 2, Kang Bin1, 2, Liu Pei1, 2, Zhong Huage4, Yu Fei1, 2   

Abstract: BACKGROUND: Chondrocytes degeneration often leads to common orthopedic diseases such as osteoarthritis, intervertebral disc degeneration and meniscus degeneration. SIRT1 gene is closely related to chondrocytes degeneration. When SIRT1 gene is knocked down in ATDC5 mouse chondrocytes, it can affect cell function via signaling pathways related to chondrocyte degeneration, and then affect the process of orthopedic diseases.
OBJECTIVE: To investigate the significantly activated signaling pathways related to chondrocyte degeneration and the expression of differential factors in ATDC5 mouse chondrocytes after SIRT1 gene knockdown.
METHODS: The lentivirus carrying SIRT1 gene or not was transfected into ATDC cells. The transfected cells were divided into control (transfected with negative control lentivirus) and experiment group (transfected with lentivirus carrying SIRT1 gene). Total RNA was extracted for quality testing. Gene chip was used to detect the expression of coding RNA in the cells. Combined with bioinformatics analysis, we identified significantly activated signaling pathways related to chondrocyte degeneration, and elaborated differentially expressed factors in some signaling pathways.
RESULTS AND CONCLUSION: Forty-two signaling pathways were significantly activated in ATDC5 cells after SIRT1 gene knockdown. Among them, we selected five signaling pathways related to chondrocyte degeneration and reported less, including hepatocyte growth factor signaling pathway (ETS1, PIK3CA, NRAS, PIK3C2A, FGFR2, ELF1, FGFR3, CDKN1A, AKT3, FRS2, PRKD3, MAP3K2; Ratio=0.139 0, Z-score=2.673), neuregulin signaling pathway   (RPS6KB1, STAT5A, MTOR, BTC, HBEGF, RNF41; Ratio=0.136 0, Z-score=2.309), insulin receptor signaling pathway (TSC1, EIF4EBP1, PRKAR2B, PPP1R12A, TSC2; Ratio=0.106 0, Z-score=2.138), chemokine receptor 4 signaling pathway (PAK4, EGR1, RHOJ, RHOB, PLCB1, GNG5; Ratio=0.097 0, Z-score=2.500), actin cytoskeleton signaling pathway (ABI2, BRK1, PIP4K2B, MYLK, IQGAP1, ARPC1A, ACTA2, EZR, SSH2, ACTG1, IQGAP3; Ratio=0.092 1, Z-score=2.236), so as to provide novel targets for treating chondrocyte degeneration related diseases.
Key words: SIRT1, ATDC5 cell, chondrocyte degeneration, gene chip, cell experiment