Abstract
Aim: Neurosyphilis patients exhibited significant expression of long noncoding RNA (lncRNA) in peripheral blood T lymphocytes. In this study, we further clarified the role of lncRNA-ENST00000421645 in the pathogenic mechanism of neurosyphilis. Methods: lncRNA-ENST00000421645 was transfected into Jurkat-E6-1 cells, namely lentivirus (Lv)-1645 cells. RNA pull-down assay, flow cytometry, RT-qPCR, ELISA (Neobioscience Technology Co Ltd, Shenzhen, China) and RNA immunoprecipitation chip assay were used to analyze the function of lncRNA-ENST00000421645. Results: The expression of IFN-γ in Lv-1645 cells was significantly increased compared to that in Jurkat-E6-1 cells stimulated by phorbol-12-myristate-13-acetate (PMA). Then, it was suggested that lncRNA-ENST00000421645 interacts with PCM1 protein. Silencing PCM1 significantly increased the level of IFN-γ in Lv-1645 cells stimulated by PMA. Conclusion: This study revealed that lncRNA-ENST00000421645 mediates the production of IFN-γ by sponging PCM1 protein after PMA stimulation.
Lay abstract
The mechanisms underlying Treponema pallidum (a type of bacterium that causes syphilis) invasion into the CNS have not yet been established. In this study, we further clarified the role of long noncoding RNA (lncRNA) in the pathogenic process causing nerve damage. The results suggested that lncRNA-ENST00000421645 interacts with an important protein named PCM1. Suppressing the expression of PCM1 significantly increased the level of IFN-γ cytokines (substances secreted by immune cells that effect other cells) with an increased level of lncRNA-ENST00000421645 when immune cells were stimulated by phorbol-12-myristate-13-acetate a specific activator of the PKC signaling enzyme involved in gene transcription pathways. This study revealed that lncRNA-ENST00000421645 mediates the production of IFN-γ by interacting with PCM1 protein.
Keywords::
W-N Liu and L-L Liu conceived and designed the experiments. Wen-Na Liu performed the experiments. W-N Liu, K-X Wu and X-T Wang analyzed the data. L-R Lin and M-L Tong contributed reagents, materials and analysis tools. W-N Liu and L-L Liu wrote the paper. All authors contributed to the article and approved the submitted version.
Financial & competing interests disclosure
This work was supported by the National Natural Science Foundation (grant numbers: 81971147, 81771312, 81471231, 81672094, 81972028, 81871729), and Xiamen Science and Technology Planning Project (grant number: 3502Z20209032). The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.
No writing assistance was utilized in the production of this manuscript.
Ethical conduct of research
This study was approved by the Ethics Committee of Zhongshan Hospital (NO. 2021-147) and complied with the Declaration of Helsinki. In addition, for investigations involving human subjects, informed consent has been obtained from the participants involved.
Availability of data & materials
The original contributions presented in the study are included in the article. Further inquiries can be directed to the corresponding author.