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RNA-Seq-based transcriptomics analysis during the photodynamic therapy of primary cells in secondary hyperparathyroidism

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Abstract

Background

The aim of this study was to identify changes in gene expression before and after 5-aminolevulinic acid-mediated photodynamic therapy (5-ALA-PDT) and to investigate the potential mechanism of 5-ALA-PDT based on ribonucleic acid sequencing (RNA-Seq) analysis.

Methods

Secondary hyperparathyroidism (SHPT) primary cells were isolated from surgically excised specimens and exposed to laser light. The transcription profiles of SHPT primary cells were identified through RNA-Seq. Differentially expressed genes (DEGs) were identified. Enrichment of functions and signaling pathway analysis were performed based on Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses. Quantitative real-time polymerase chain reaction (RT-qPCR) and western blot analysis were used to validate genes based on RNA-Seq results.

Results

In total, 1320 DEGs were identified, of which 1019 genes were upregulated and 301 genes were downregulated. GO and KEGG pathway analyses identified significantly enriched pathways in DEGs, including TGF beta in extracellular matrix (ECM), negative regulation of triglyceride biosynthetic process, protein heterodimerization activity, systemic lupus erythematosus, ECM–receptor interaction, focal adhesion and protein digestion and absorption. Protein–protein interaction (PPI) network analyses identified potential heat shock protein (HSP) interactions among the DEGs. Eight HSP genes were also identified that were most likely involved in 5-ALA-PDT, which were further validated by RT-qPCR and western blotting.

Conclusions

The findings of this descriptive study reveal changes in the transcriptome profile during 5-ALA-PDT, suggesting that gene expression and mutation, signaling pathways, and the molecular network are altered in SHPT primary cells. The above findings provide new insight for further studies on the mechanisms underlying 5-ALA-PDT in SHPT.

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Data availability

The data that support the findings of this study are available from the corresponding author upon reasonable request. The data are not publicly available due to privacy.

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Acknowledgements

This research was supported by the National Natural Science Foundation of China (Grants 82270834, 81873640 and 82000756) and the Fundamental Research Funds for the Central Universities of Central South University (Grant 202ZTS0950).

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  1. Ying Wen and Liyun Zeng have contributed equally to this work.

Authors and Affiliations

  1. Department of General Surgery, The Second Xiangya Hospital, Central South University, No. 139, Renmin Central Road, Changsha, 410011, People’s Republic of China

    Ying Wen, Liyun Zeng, Qitong Chen, Yitong Li, Mengdie Fu, Zixin Wang, Qiongyan Zou & Wenjun Yi

  2. Department of Nephrology, The Second Xiangya Hospital, Central South University, Changsha, People’s Republic of China

    Hong Liu & Xiejia Li

  3. Department of General Surgery, Xiangya Hospital, Central South University, Changsha, People’s Republic of China

    Peng Huang

  4. Department of General Surgery, The Third Xiangya Hospital, Central South University, Changsha, People’s Republic of China

    Wei Wu

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Ying Wen, Liyun Zeng, Qitong Chen, Yitong Li, Mengdie Fu, Zixin Wang, Hong Liu, Xiejia Li, Peng Huang, Wei Wu, Qiongyan Zou, Wenjun Yi has been approved by all authors.

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Wen, Y., Zeng, L., Chen, Q. et al. RNA-Seq-based transcriptomics analysis during the photodynamic therapy of primary cells in secondary hyperparathyroidism. Photochem Photobiol Sci 22, 905–917 (2023). https://doi.org/10.1007/s43630-023-00361-0

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