Quantitative analysis of transcriptome and proteome of knee joint synovial cells with osteopontin knockdown

doi:10.12307/2022.351

Chinese Journal of Tissue Engineering Research ›› 2022, Vol. 26 ›› Issue (11): 1680-1685.doi: 10.12307/2022.351

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Quantitative analysis of transcriptome and proteome of knee joint synovial cells with osteopontin knockdown

Xu Renjie1, 2, Zhong Qiao1, Liu Yubo1, Yu Xiao1, Yan Yongqing3, Saijilafu2, Yang Huilin2, Chen Guangxiang1

1Department of Orthopedics, Suzhou Municipal Hospital/The Affiliated Hospital of Nanjing Medical University, Suzhou 215000, Jiangsu Province, China; 2Orthopedic Institute, Soochow University, Suzhou 215000, Jiangsu Province, China; 3Ningbo Huamei Hospital, University of Chinese Academy of Sciences, Ningbo 315000, Zhejiang Province, China

Received:2021-01-05 Revised:2021-01-30 Accepted:2021-08-03 Online:2022-04-18 Published:2021-12-11
Contact: Chen Guangxiang, MD, Master’s supervisor, Department of Orthopedics, Suzhou Municipal Hospital/The Affiliated Hospital of Nanjing Medical University, Suzhou 215000, Jiangsu Province, China Yang Huilin, MD, Doctoral supervisor, Orthopedic Institute, Soochow University, Suzhou 215000, Jiangsu Province, China
About author:Xu Renjie, MD, Chief physician, Department of Orthopedics, Suzhou Municipal Hospital/The Affiliated Hospital of Nanjing Medical University, Suzhou 215000, Jiangsu Province, China Zhong Qiao, MD, Associate chief technician, Department of Orthopedics, Suzhou Municipal Hospital/The Affiliated Hospital of Nanjing Medical University, Suzhou 215000, Jiangsu Province, China Xu Renjie and Zhong Qiao contributed equally to this work.
Supported by:
the National Natural Science Foundation of China (General Project), No. 81971995 (to ZQ); Jiangsu Basic Research Program-General Project, No. BK20191179 (to ZQ); Scientific Research Project of Jiangsu Provincial Health and Family Planning Commission, No. LGY2017007 (to XRJ); Suzhou Medical and Health Application Basic Research Project, No. SYSD2016110 (to XRJ); Zhejiang Provincial Medicine and Health Scientific Project, No. 2019319060 (to YYQ)

Abstract

Abstract: BACKGROUND: The mRNA and protein levels of osteopontin in articular cartilage and synovial fluid of patients with osteoarthritis are significantly higher than those of normal controls. However, the role and specific mechanism of osteopontin in the development of knee osteoarthritis have not been clarified.
OBJECTIVE: To investigate the level of osteopontin in synovial cells of knee joint osteoarthritis patients and its relationship with the severity of osteoarthritis, and to further elucidate the mechanism that affects the progression of osteoarthritis through transcriptome and proteome sequencing.
METHODS: A total of 42 patients with primary knee osteoarthritis diagnosed and treated in 2019 were divided into unicompartment knee osteoarthritis group and bicompartment knee osteoarthritis group based on the involvement of the tibiofemoral joint. Synovial and joint fluid specimens were harvested. PCR method was used to measure the abundance of osteopontin gene expression in synovial specimens, and ELISA method was used to measure osteopontin protein level in synovial fluid specimens. Enzymatic digestion method was used to obtain and culture human knee osteoarthritis synovial fibroblast-like cells. After osteopontin-siRNA intervention for 24 hours, the samples were tested for transcriptome and proteome.
RESULTS AND CONCLUSION: Compared with the unicompartment knee osteoarthritis group, the osteopontin levels in the synovium and synovial fluid of the bicompartment knee osteoarthritis group increased significantly (P < 0.05). In in vitro cultured human knee osteoarthritis synovial fibroblast-like cells, after osteopontin knockdown by siRNA, quantitative studies of the transcriptome and proteome identified 14 662 transcripts and 3 608 proteins, respectively. Among them, the transcriptome and proteome analysis results of five proteins showed a simultaneous decrease, and the transcriptome and proteome analysis results of two proteins showed a simultaneous increase. To conclude, osteopontin has an adverse effect on chondrocytes. The mechanism may be through the pro-inflammatory effect and the negative effect of insulin-like growth factor-binding protein 3 on cartilage.

Key words: knee joint, osteoarthritis, osteopontin, synovial cells, transcriptome, proteome, mechanism

CLC Number:

Xu Renjie, Zhong Qiao, Liu Yubo, Yu Xiao, Yan Yongqing, Saijilafu, Yang Huilin, Chen Guangxiang. Quantitative analysis of transcriptome and proteome of knee joint synovial cells with osteopontin knockdown[J]. Chinese Journal of Tissue Engineering Research, 2022, 26(11): 1680-1685.

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References

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Quantitative analysis of transcriptome and proteome of knee joint synovial cells with osteopontin knockdown

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