PKR associates with 4.1R to promote anchorage-independent growth of hepatocellular carcinoma and lead to poor prognosis
Abstract RNA-dependent protein kinase (PKR) may have a positive regulatory role in controlling tumor growth and progression in hepatocellular carcinoma (HCC). However, the downstream substrates and the molecular mechanism of PKR in the growth and progression of HCC have not been clarified. In this s...
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Nature Portfolio
2024-11-01
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| Online Access: | https://doi.org/10.1038/s41598-024-75142-5 |
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| author | Yusuke Okujima Takao Watanabe Takeshi Ito Yasumichi Inoue Yutaka Kasai Yusuke Imai Yoshiko Nakamura Mitsuhito Koizumi Osamu Yoshida Yoshio Tokumoto Masashi Hirooka Masanori Abe Ryosuke Kawakami Takashi Saitou Takeshi Imamura Yoshinori Murakami Yoichi Hiasa |
| author_facet | Yusuke Okujima Takao Watanabe Takeshi Ito Yasumichi Inoue Yutaka Kasai Yusuke Imai Yoshiko Nakamura Mitsuhito Koizumi Osamu Yoshida Yoshio Tokumoto Masashi Hirooka Masanori Abe Ryosuke Kawakami Takashi Saitou Takeshi Imamura Yoshinori Murakami Yoichi Hiasa |
| author_sort | Yusuke Okujima |
| collection | DOAJ |
| description | Abstract RNA-dependent protein kinase (PKR) may have a positive regulatory role in controlling tumor growth and progression in hepatocellular carcinoma (HCC). However, the downstream substrates and the molecular mechanism of PKR in the growth and progression of HCC have not been clarified. In this study, mass spectrometry analysis was performed with immunoprecipitated samples, and 4.1R was identified as a protein that binds to PKR. In transfected COS7 cells, an immunoprecipitation experiment showed that 4.1R binds to wild-type PKR, but not to a kinase-deficient mutant PKR, suggesting that PKR binds to 4.1R in a kinase activity-dependent manner. In HCC cell lines, HuH7 and HepG2, the expression level of 4.1R protein was shown to be regulated by protein expression and activation of PKR. Interestingly, high expression of 4.1R, as well as PKR, is associated with a worse prognosis in HCC. PKR increased HCC cell growth in both anchorage-dependent and anchorage-independent manners, whereas 4.1R was involved in HCC cell growth only in an anchorage-independent manner, not in an anchorage-dependent manner. The rescue experiment indicated that increased anchorage-independent growth of HCC cells by PKR might be caused by 4.1R. In conclusion, PKR associates with 4.1R and promotes anchorage-independent growth of HCC. The PKR-4.1R axis might be a new therapeutic target in HCC. |
| format | Article |
| id | doaj-art-dd42980cfe6a449483d6c8e426e3ad7c |
| institution | Kabale University |
| issn | 2045-2322 |
| language | English |
| publishDate | 2024-11-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj-art-dd42980cfe6a449483d6c8e426e3ad7c2024-11-17T12:25:30ZengNature PortfolioScientific Reports2045-23222024-11-0114111410.1038/s41598-024-75142-5PKR associates with 4.1R to promote anchorage-independent growth of hepatocellular carcinoma and lead to poor prognosisYusuke Okujima0Takao Watanabe1Takeshi Ito2Yasumichi Inoue3Yutaka Kasai4Yusuke Imai5Yoshiko Nakamura6Mitsuhito Koizumi7Osamu Yoshida8Yoshio Tokumoto9Masashi Hirooka10Masanori Abe11Ryosuke Kawakami12Takashi Saitou13Takeshi Imamura14Yoshinori Murakami15Yoichi Hiasa16Department of Gastroenterology and Metabology, Graduate School of Medicine, Ehime UniversityDepartment of Gastroenterology and Metabology, Graduate School of Medicine, Ehime UniversityDivision of Molecular Pathology, The Institute of Medical Science, The University of TokyoDepartment of Cell Signaling, Graduate School of Pharmaceutical Sciences, Nagoya City UniversityDivision of Molecular Pathology, The Institute of Medical Science, The University of TokyoDepartment of Gastroenterology and Metabology, Graduate School of Medicine, Ehime UniversityDepartment of Gastroenterology and Metabology, Graduate School of Medicine, Ehime UniversityDepartment of Gastroenterology and Metabology, Graduate School of Medicine, Ehime UniversityDepartment of Gastroenterology and Metabology, Graduate School of Medicine, Ehime UniversityDepartment of Gastroenterology and Metabology, Graduate School of Medicine, Ehime UniversityDepartment of Gastroenterology and Metabology, Graduate School of Medicine, Ehime UniversityDepartment of Gastroenterology and Metabology, Graduate School of Medicine, Ehime UniversityDepartment of Molecular Medicine for Pathogenesis, Graduate School of Medicine, Ehime UniversityDepartment of Molecular Medicine for Pathogenesis, Graduate School of Medicine, Ehime UniversityDepartment of Molecular Medicine for Pathogenesis, Graduate School of Medicine, Ehime UniversityDivision of Molecular Pathology, The Institute of Medical Science, The University of TokyoDepartment of Gastroenterology and Metabology, Graduate School of Medicine, Ehime UniversityAbstract RNA-dependent protein kinase (PKR) may have a positive regulatory role in controlling tumor growth and progression in hepatocellular carcinoma (HCC). However, the downstream substrates and the molecular mechanism of PKR in the growth and progression of HCC have not been clarified. In this study, mass spectrometry analysis was performed with immunoprecipitated samples, and 4.1R was identified as a protein that binds to PKR. In transfected COS7 cells, an immunoprecipitation experiment showed that 4.1R binds to wild-type PKR, but not to a kinase-deficient mutant PKR, suggesting that PKR binds to 4.1R in a kinase activity-dependent manner. In HCC cell lines, HuH7 and HepG2, the expression level of 4.1R protein was shown to be regulated by protein expression and activation of PKR. Interestingly, high expression of 4.1R, as well as PKR, is associated with a worse prognosis in HCC. PKR increased HCC cell growth in both anchorage-dependent and anchorage-independent manners, whereas 4.1R was involved in HCC cell growth only in an anchorage-independent manner, not in an anchorage-dependent manner. The rescue experiment indicated that increased anchorage-independent growth of HCC cells by PKR might be caused by 4.1R. In conclusion, PKR associates with 4.1R and promotes anchorage-independent growth of HCC. The PKR-4.1R axis might be a new therapeutic target in HCC.https://doi.org/10.1038/s41598-024-75142-5eIF-2 kinaseErythrocyte membrane band 4.1 proteinLiver neoplasmsCell linePrognosis |
| spellingShingle | Yusuke Okujima Takao Watanabe Takeshi Ito Yasumichi Inoue Yutaka Kasai Yusuke Imai Yoshiko Nakamura Mitsuhito Koizumi Osamu Yoshida Yoshio Tokumoto Masashi Hirooka Masanori Abe Ryosuke Kawakami Takashi Saitou Takeshi Imamura Yoshinori Murakami Yoichi Hiasa PKR associates with 4.1R to promote anchorage-independent growth of hepatocellular carcinoma and lead to poor prognosis Scientific Reports eIF-2 kinase Erythrocyte membrane band 4.1 protein Liver neoplasms Cell line Prognosis |
| title | PKR associates with 4.1R to promote anchorage-independent growth of hepatocellular carcinoma and lead to poor prognosis |
| title_full | PKR associates with 4.1R to promote anchorage-independent growth of hepatocellular carcinoma and lead to poor prognosis |
| title_fullStr | PKR associates with 4.1R to promote anchorage-independent growth of hepatocellular carcinoma and lead to poor prognosis |
| title_full_unstemmed | PKR associates with 4.1R to promote anchorage-independent growth of hepatocellular carcinoma and lead to poor prognosis |
| title_short | PKR associates with 4.1R to promote anchorage-independent growth of hepatocellular carcinoma and lead to poor prognosis |
| title_sort | pkr associates with 4 1r to promote anchorage independent growth of hepatocellular carcinoma and lead to poor prognosis |
| topic | eIF-2 kinase Erythrocyte membrane band 4.1 protein Liver neoplasms Cell line Prognosis |
| url | https://doi.org/10.1038/s41598-024-75142-5 |
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