1.Screening and biologically functional analysis of differentially expressed genes for neuropathic pain
Peng SHEN ; Xinping CHAI ; Yutong LI
Chinese Journal of Rehabilitation Medicine 2024;39(10):1430-1435,1442
Objective:To investigate differentially expressed genes(DEGs)for neuropathic pain between sciatica and healthy controls. Method:The microarray dataset GSE150408 were downloaded from gene expression database(gene expression omnibus database,GEO).The DEGs were screened using the limma V3.42.0(linear models for microarray da-ta)package of the R software program(version 3.5.0).GEO terms and Kyoto encyclopedia of genes and ge-nomes(KEGG)pathway enrichment analysis of DEGs were automatically completed and visualized by the clus-terProiler V3.14.0.STRING was searched to identify and predict interactions between genes or proteins,to con-struct the protein to protein interaction. Result:A total of 424 DEGs were screened,including 233 up-regulated genes and 191 down-regulated genes.The result of GO enrichment indicated that for biological process,DEGs were significantly enriched in de-fense response,regulation of immune system process,cell activation,leukocyte activation.Regarding cell com-ponents,DEGs were significantly enriched in vesicles,secretory granules,tertiary granule.For molecular func-tion,DEGs were significantly enriched in immune receptor activation,complement receptor activity and MAP kinase phosphatase activity.The result of KEGG pathway-enrichment indicated that DEGs were mainly enriched in transcriptional misregulation in cancer.Moreover,the GSEA analysis indicated that the most significant en-riched gene sets included endocytosis,Epstein-Barr virus infection.PPI analysis were performed to explore the potential function of the DEGs.It showed that the degree of FCGR1A was the highest in up-regulated proteins and the highest in down-regulated proteins.These two proteins were the key nodes. Conclusion:It showed an inflammatory and immune characteristic in peripheral blood for neuropathic pain.AZU1,BPI,FCGR1A and SPTBN2 were the key genes in neuropathic pain.
2.Circ_0068655 Promotes Cardiomyocyte Apoptosis via miR-498/ PAWR Axis
Qiaoying CHAI ; Mingqi ZHENG ; Zheng WANG ; Mei WE ; Yajuan YIN ; Fangfang MA ; Xinping LI ; Haijun ZHANG ; Gang LIU
Tissue Engineering and Regenerative Medicine 2020;17(5):659-670
BACKGROUND:
The cardiomyocyte apoptosis is considered as one of major contributions to cardiac remodeling after myocardial infarction (MI). Numerous studies find that circular RNAs (circRNAs) play pivotal roles in a variety of biological functions. However, the role of circ_0068655 in MI and human induced pluripotent stem-derived cardiomyocytes (HCMs) remains unknown.
METHODS:
The expression of circ_0068655, miR-498, and PRKC apoptosis WT1 regulator (PAWR) in human MI heart tissues and hypoxia subjected HCMs was evaluated with qRT-PCR and Western blot. The effects of circ_0068655 on hypoxia-induced apoptotic death and cell migration in HCMs were evaluated with qRT-PCR, cell viability, cell death ELISA (POD), and Caspase-3 activity assay, and Trans-well assay, respectively. Furthermore, luciferase assay, qRT-PCR, biotin-labeled miRNA pulldown assay, and Western blot were employed in the functional studies.
RESULTS:
We found that the expression of circ_0068655 and PAWR was enhanced in MI patients and hypoxia subjected HCMs; by contrast, the expression of miR-498 decreased. Inhibited expression of circ_0068655 in HMCs counteracted hypoxia-induced apoptotic death and impaired cell migration, in sharp contrast to circ_0068655 knockdown. We identified that circ_0068655 sponged an endogenous miR-498 to sequester and inhibit its activity, leading to the increased PAWR expression.
CONCLUSIONS
Our findings reveal that the expression of circ_0068655 can promote cardiomyocyte apoptosis through the modulation of miR-498-PAWR axis in vitro, which highlights the diagnostic and therapeutic value of circ_0068655 in patients with MI.
3.Circ_0068655 Promotes Cardiomyocyte Apoptosis via miR-498/ PAWR Axis
Qiaoying CHAI ; Mingqi ZHENG ; Zheng WANG ; Mei WE ; Yajuan YIN ; Fangfang MA ; Xinping LI ; Haijun ZHANG ; Gang LIU
Tissue Engineering and Regenerative Medicine 2020;17(5):659-670
BACKGROUND:
The cardiomyocyte apoptosis is considered as one of major contributions to cardiac remodeling after myocardial infarction (MI). Numerous studies find that circular RNAs (circRNAs) play pivotal roles in a variety of biological functions. However, the role of circ_0068655 in MI and human induced pluripotent stem-derived cardiomyocytes (HCMs) remains unknown.
METHODS:
The expression of circ_0068655, miR-498, and PRKC apoptosis WT1 regulator (PAWR) in human MI heart tissues and hypoxia subjected HCMs was evaluated with qRT-PCR and Western blot. The effects of circ_0068655 on hypoxia-induced apoptotic death and cell migration in HCMs were evaluated with qRT-PCR, cell viability, cell death ELISA (POD), and Caspase-3 activity assay, and Trans-well assay, respectively. Furthermore, luciferase assay, qRT-PCR, biotin-labeled miRNA pulldown assay, and Western blot were employed in the functional studies.
RESULTS:
We found that the expression of circ_0068655 and PAWR was enhanced in MI patients and hypoxia subjected HCMs; by contrast, the expression of miR-498 decreased. Inhibited expression of circ_0068655 in HMCs counteracted hypoxia-induced apoptotic death and impaired cell migration, in sharp contrast to circ_0068655 knockdown. We identified that circ_0068655 sponged an endogenous miR-498 to sequester and inhibit its activity, leading to the increased PAWR expression.
CONCLUSIONS
Our findings reveal that the expression of circ_0068655 can promote cardiomyocyte apoptosis through the modulation of miR-498-PAWR axis in vitro, which highlights the diagnostic and therapeutic value of circ_0068655 in patients with MI.