Transcriptome analysis of primary human oral keratinocytes stimulated with Prevotella melaninogenica
Objective To investigate the transcriptomic changes in primary human oral keratinocytes (pHOKs) after coculture with Prevotella melaninogenica (P.m) and to verify the changes in human oral keratinocyte (HOK) cell lines. Methods pHOK was isolated and cocultured with P.m for 0, 4 and 24 h. Total RNA w...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | zho |
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Editorial Department of Journal of Prevention and Treatment for Stomatological Diseases
2022-09-01
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| Series: | 口腔疾病防治 |
| Subjects: | |
| Online Access: | http://www.kqjbfz.com/CN/10.12016/j.issn.2096-1456.2022.09.002 |
| _version_ | 1828803756721963008 |
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| author | GUO Yiting HAN Wenhao XU Pan SHAO Ruru HE Yuan |
| author_facet | GUO Yiting HAN Wenhao XU Pan SHAO Ruru HE Yuan |
| author_sort | GUO Yiting |
| collection | DOAJ |
| description | Objective To investigate the transcriptomic changes in primary human oral keratinocytes (pHOKs) after coculture with Prevotella melaninogenica (P.m) and to verify the changes in human oral keratinocyte (HOK) cell lines. Methods pHOK was isolated and cocultured with P.m for 0, 4 and 24 h. Total RNA was extracted, a gene library was constructed, transcriptional sequencing was performed, differentially expressed genes (DEGs) were analyzed, gene ontology (GO) pathway analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were performed, and the validation of DEGs was performed by qRT-PCR and Western Blot in the HOK and P.m coculture cell model. Results 1 788 DEGs were detected between the 4 h group and control group, including upregulated DEGs such as lymphocyte cytosolic protein 1(LCP1), keratin 7 (KRT7) and Cilia and flagella associated protein 251(CFAP251) and downregulated DEGs such as FERM, ARH/RhoGEF and Pleckstrin domain protein 1 (FARP1), WW domain containing transcription regulator 1(WWTR1) and Discoidin, CUB and LCCL domain-containing protein 2 (DCBLD2). 1 832 DEGs were detected between the 24 h group and control group, including upregulated DEGs such as LCP1, complement C1s(C1S), kynureninase (KYNU) and downregulated DEGs such as phosphoserine aminotransferase 1 (PSAT1), FARP1 and FKBP prolyl isomerase 10 (FKBP10). There were 1 090 common differentially expressed genes (cDEGs) in the 4 h and 24 h groups, including LCP1, KYNU and long intergenic nonprotein coding RNA 958 (LINC00958). The GO pathways were mainly enriched in response to lipopolysaccharide and the molecules of bacterial origin and apical part of the cell. KEGG pathway analysis revealed enrichment in the interleukin-17 (IL-17) signaling pathway, tumor necrosis factor (TNF) signaling pathway, Toll-like receptor (TLR) pathway, etc. We verified the expression of a cDEG, Myosin1B (MYO1B), and qRT-PCR and Western Blot analysis showed that MYO1B expression was significantly upregulated between the control group and the P.m cocultured group (P<0.001), and its expression followed a time-dependent and concentration-dependent manner. Conclusion P.m played an important role in the transcriptome of oral keratinocytes. |
| first_indexed | 2024-12-12T07:28:55Z |
| format | Article |
| id | doaj.art-694534236a8b4514830f56c73a746b83 |
| institution | Directory Open Access Journal |
| issn | 2096-1456 |
| language | zho |
| last_indexed | 2024-12-12T07:28:55Z |
| publishDate | 2022-09-01 |
| publisher | Editorial Department of Journal of Prevention and Treatment for Stomatological Diseases |
| record_format | Article |
| series | 口腔疾病防治 |
| spelling | doaj.art-694534236a8b4514830f56c73a746b832022-12-22T00:33:04ZzhoEditorial Department of Journal of Prevention and Treatment for Stomatological Diseases口腔疾病防治2096-14562022-09-0130962062910.12016/j.issn.2096-1456.2022.09.002Transcriptome analysis of primary human oral keratinocytes stimulated with Prevotella melaninogenicaGUO Yiting0 HAN Wenhao1XU Pan2SHAO Ruru3 HE Yuan4School of Stomatology, Tongji University Department of Gastroenterology, Shanghai 10th People’s Hospital & School of Life Sciences and Technology, Tongji UniversitySchool of Stomatology, Tongji UniversitySchool of Stomatology, Tongji UniversitySchool of Stomatology, Tongji UniversityObjective To investigate the transcriptomic changes in primary human oral keratinocytes (pHOKs) after coculture with Prevotella melaninogenica (P.m) and to verify the changes in human oral keratinocyte (HOK) cell lines. Methods pHOK was isolated and cocultured with P.m for 0, 4 and 24 h. Total RNA was extracted, a gene library was constructed, transcriptional sequencing was performed, differentially expressed genes (DEGs) were analyzed, gene ontology (GO) pathway analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were performed, and the validation of DEGs was performed by qRT-PCR and Western Blot in the HOK and P.m coculture cell model. Results 1 788 DEGs were detected between the 4 h group and control group, including upregulated DEGs such as lymphocyte cytosolic protein 1(LCP1), keratin 7 (KRT7) and Cilia and flagella associated protein 251(CFAP251) and downregulated DEGs such as FERM, ARH/RhoGEF and Pleckstrin domain protein 1 (FARP1), WW domain containing transcription regulator 1(WWTR1) and Discoidin, CUB and LCCL domain-containing protein 2 (DCBLD2). 1 832 DEGs were detected between the 24 h group and control group, including upregulated DEGs such as LCP1, complement C1s(C1S), kynureninase (KYNU) and downregulated DEGs such as phosphoserine aminotransferase 1 (PSAT1), FARP1 and FKBP prolyl isomerase 10 (FKBP10). There were 1 090 common differentially expressed genes (cDEGs) in the 4 h and 24 h groups, including LCP1, KYNU and long intergenic nonprotein coding RNA 958 (LINC00958). The GO pathways were mainly enriched in response to lipopolysaccharide and the molecules of bacterial origin and apical part of the cell. KEGG pathway analysis revealed enrichment in the interleukin-17 (IL-17) signaling pathway, tumor necrosis factor (TNF) signaling pathway, Toll-like receptor (TLR) pathway, etc. We verified the expression of a cDEG, Myosin1B (MYO1B), and qRT-PCR and Western Blot analysis showed that MYO1B expression was significantly upregulated between the control group and the P.m cocultured group (P<0.001), and its expression followed a time-dependent and concentration-dependent manner. Conclusion P.m played an important role in the transcriptome of oral keratinocytes.http://www.kqjbfz.com/CN/10.12016/j.issn.2096-1456.2022.09.002oral lichen planusprevotella melaninogenicarna-seqmyosin1bepithelial barrier functioninterleukin-17 signaling pathwaytumor necrosis factor signaling pathwaytoll-like receptor pathwayapical part of cell |
| spellingShingle | GUO Yiting HAN Wenhao XU Pan SHAO Ruru HE Yuan Transcriptome analysis of primary human oral keratinocytes stimulated with Prevotella melaninogenica 口腔疾病防治 oral lichen planus prevotella melaninogenica rna-seq myosin1b epithelial barrier function interleukin-17 signaling pathway tumor necrosis factor signaling pathway toll-like receptor pathway apical part of cell |
| title | Transcriptome analysis of primary human oral keratinocytes stimulated with Prevotella melaninogenica |
| title_full | Transcriptome analysis of primary human oral keratinocytes stimulated with Prevotella melaninogenica |
| title_fullStr | Transcriptome analysis of primary human oral keratinocytes stimulated with Prevotella melaninogenica |
| title_full_unstemmed | Transcriptome analysis of primary human oral keratinocytes stimulated with Prevotella melaninogenica |
| title_short | Transcriptome analysis of primary human oral keratinocytes stimulated with Prevotella melaninogenica |
| title_sort | transcriptome analysis of primary human oral keratinocytes stimulated with prevotella melaninogenica |
| topic | oral lichen planus prevotella melaninogenica rna-seq myosin1b epithelial barrier function interleukin-17 signaling pathway tumor necrosis factor signaling pathway toll-like receptor pathway apical part of cell |
| url | http://www.kqjbfz.com/CN/10.12016/j.issn.2096-1456.2022.09.002 |
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