ER resident protein 44 promotes malignant phenotype in nasopharyngeal carcinoma through the interaction with ATP citrate lyase
Abstract Background Nasopharyngeal carcinoma (NPC) is one of the most common malignancy in head and neck. With the development of treatments, the prognosis has improved these years, but metastasis is still the main cause of treatment failure. The endoplasmic reticulum (ER) resident protein 44 is a U...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2021-02-01
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| Series: | Journal of Translational Medicine |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12967-020-02694-1 |
| _version_ | 1818415752099659776 |
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| author | Hui Tian Si Shi Bo You Qicheng Zhang Miao Gu Yiwen You |
| author_facet | Hui Tian Si Shi Bo You Qicheng Zhang Miao Gu Yiwen You |
| author_sort | Hui Tian |
| collection | DOAJ |
| description | Abstract Background Nasopharyngeal carcinoma (NPC) is one of the most common malignancy in head and neck. With the development of treatments, the prognosis has improved these years, but metastasis is still the main cause of treatment failure. The endoplasmic reticulum (ER) resident protein 44 is a UPR-induced ER protein of the protein disulphide isomerase (PDI) family. This study investigated the role of ERp44 in NPC progression. Methods Firstly, immunohistochemistry, western blot and qRT-PCR were used to investigate the expression of ERp44 in NPC samples and cell lines. We analyzed 44 NPC samples for ERp44 expression and investigated the association between its expression level with clinicopathologic parameters. Then we took CCK8, Transwell migration assay and used the zebrafish model to access the role of ERp44 on the malignant phenotype in NPC cells. Secondly, we used co-IP to gain the proteins that interact with ERp44 and took proteomic analysis. Furthermore, we successfully constructed the mutant variants of ERp44 and found the interaction domain with ATP citrate lyase(ACLY). Lastly, we subcutaneously injected NPC cells into nude mice and took immunohistochemistry to exam the expression of ACLY and ERp44. Then we used western blot to detect the expression level of epithelial-mesenchymal transition (EMT) markers. Results In the present study, we found ERp44 was elevated in NPC tissues and correlated with clinical stages and survive state of the patients. In vitro, the downregulation of ERp44 in NPC cells (CNE2, 5-8F) could suppress cells proliferation and migration. After that, we recognized that ACLY might be a potential target that could interact with ERp44. We further constructed the mutant variants of ERp44 and found the interaction domain with ACLY. The promotion of ERp44 on cell migration could be inhibited when ACLY was knocked down. More importantly, we also observed that the interaction of ERp44 with ACLY, especially the thioredoxin region in ERp44 play a vital role in regulating EMT. Lastly, we found ERp44 was positively correlated with the expression of ACLY and could promote NPC cells growth in nude mice. Conclusion Our data indicated that ERp44 participates in promoting NPC progression through the interaction with ACLY and regulation of EMT. |
| first_indexed | 2024-12-14T11:39:59Z |
| format | Article |
| id | doaj.art-94f694c1b86d4d07a4828f3addfce81a |
| institution | Directory Open Access Journal |
| issn | 1479-5876 |
| language | English |
| last_indexed | 2024-12-14T11:39:59Z |
| publishDate | 2021-02-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Translational Medicine |
| spelling | doaj.art-94f694c1b86d4d07a4828f3addfce81a2022-12-21T23:02:53ZengBMCJournal of Translational Medicine1479-58762021-02-0119111310.1186/s12967-020-02694-1ER resident protein 44 promotes malignant phenotype in nasopharyngeal carcinoma through the interaction with ATP citrate lyaseHui Tian0Si Shi1Bo You2Qicheng Zhang3Miao Gu4Yiwen You5Department of Otorhinolaryngology Head and Neck Surgery, Affiliated Hospital of Nantong UniversityDepartment of Otorhinolaryngology Head and Neck Surgery, Affiliated Hospital of Nantong UniversityDepartment of Otorhinolaryngology Head and Neck Surgery, Affiliated Hospital of Nantong UniversityDepartment of Otorhinolaryngology Head and Neck Surgery, Affiliated Hospital of Nantong UniversityDepartment of Otorhinolaryngology Head and Neck Surgery, Affiliated Hospital of Nantong UniversityDepartment of Otorhinolaryngology Head and Neck Surgery, Affiliated Hospital of Nantong UniversityAbstract Background Nasopharyngeal carcinoma (NPC) is one of the most common malignancy in head and neck. With the development of treatments, the prognosis has improved these years, but metastasis is still the main cause of treatment failure. The endoplasmic reticulum (ER) resident protein 44 is a UPR-induced ER protein of the protein disulphide isomerase (PDI) family. This study investigated the role of ERp44 in NPC progression. Methods Firstly, immunohistochemistry, western blot and qRT-PCR were used to investigate the expression of ERp44 in NPC samples and cell lines. We analyzed 44 NPC samples for ERp44 expression and investigated the association between its expression level with clinicopathologic parameters. Then we took CCK8, Transwell migration assay and used the zebrafish model to access the role of ERp44 on the malignant phenotype in NPC cells. Secondly, we used co-IP to gain the proteins that interact with ERp44 and took proteomic analysis. Furthermore, we successfully constructed the mutant variants of ERp44 and found the interaction domain with ATP citrate lyase(ACLY). Lastly, we subcutaneously injected NPC cells into nude mice and took immunohistochemistry to exam the expression of ACLY and ERp44. Then we used western blot to detect the expression level of epithelial-mesenchymal transition (EMT) markers. Results In the present study, we found ERp44 was elevated in NPC tissues and correlated with clinical stages and survive state of the patients. In vitro, the downregulation of ERp44 in NPC cells (CNE2, 5-8F) could suppress cells proliferation and migration. After that, we recognized that ACLY might be a potential target that could interact with ERp44. We further constructed the mutant variants of ERp44 and found the interaction domain with ACLY. The promotion of ERp44 on cell migration could be inhibited when ACLY was knocked down. More importantly, we also observed that the interaction of ERp44 with ACLY, especially the thioredoxin region in ERp44 play a vital role in regulating EMT. Lastly, we found ERp44 was positively correlated with the expression of ACLY and could promote NPC cells growth in nude mice. Conclusion Our data indicated that ERp44 participates in promoting NPC progression through the interaction with ACLY and regulation of EMT.https://doi.org/10.1186/s12967-020-02694-1ERp44Nasopharyngeal carcinomaMetastasisACLYEMT |
| spellingShingle | Hui Tian Si Shi Bo You Qicheng Zhang Miao Gu Yiwen You ER resident protein 44 promotes malignant phenotype in nasopharyngeal carcinoma through the interaction with ATP citrate lyase Journal of Translational Medicine ERp44 Nasopharyngeal carcinoma Metastasis ACLY EMT |
| title | ER resident protein 44 promotes malignant phenotype in nasopharyngeal carcinoma through the interaction with ATP citrate lyase |
| title_full | ER resident protein 44 promotes malignant phenotype in nasopharyngeal carcinoma through the interaction with ATP citrate lyase |
| title_fullStr | ER resident protein 44 promotes malignant phenotype in nasopharyngeal carcinoma through the interaction with ATP citrate lyase |
| title_full_unstemmed | ER resident protein 44 promotes malignant phenotype in nasopharyngeal carcinoma through the interaction with ATP citrate lyase |
| title_short | ER resident protein 44 promotes malignant phenotype in nasopharyngeal carcinoma through the interaction with ATP citrate lyase |
| title_sort | er resident protein 44 promotes malignant phenotype in nasopharyngeal carcinoma through the interaction with atp citrate lyase |
| topic | ERp44 Nasopharyngeal carcinoma Metastasis ACLY EMT |
| url | https://doi.org/10.1186/s12967-020-02694-1 |
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