NPRL2 down‐regulation facilitates the growth of hepatocellular carcinoma via the mTOR pathway and autophagy suppression
Abstract Hepatocellular carcinoma (HCC) is a highly invasive malignancy. Recently, GATOR1 (Gap Activity TOward Rags 1) complexes have been shown to play an important role in regulating tumor growth. NPRL2 is a critical component of the GATOR1 complex. Therefore, this study used NPRL2 knockdown to in...
| Main Authors: | , , , , , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Wolters Kluwer Health/LWW
2022-12-01
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| Series: | Hepatology Communications |
| Online Access: | https://doi.org/10.1002/hep4.2019 |
| _version_ | 1797759657563914240 |
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| author | Ya‐Chin Wang Ming‐Chao Tsai Yaw‐Sen Chen Pei‐Min Hsieh Chao‐Ming Hung Hung‐Yu Lin Yao‐Chun Hsu Jen‐Hao Yeh Pojen Hsiao Yu‐Cheih Su Ching‐Hou Ma Chih‐Yuan Lee Chih‐Che Lin Chih‐Wen Shu Yu‐Chan Li Mei‐Hsing Tsai James Yu Lin Wei‐Hao Peng Ming‐Lung Yu Chih‐Wen Lin |
| author_facet | Ya‐Chin Wang Ming‐Chao Tsai Yaw‐Sen Chen Pei‐Min Hsieh Chao‐Ming Hung Hung‐Yu Lin Yao‐Chun Hsu Jen‐Hao Yeh Pojen Hsiao Yu‐Cheih Su Ching‐Hou Ma Chih‐Yuan Lee Chih‐Che Lin Chih‐Wen Shu Yu‐Chan Li Mei‐Hsing Tsai James Yu Lin Wei‐Hao Peng Ming‐Lung Yu Chih‐Wen Lin |
| author_sort | Ya‐Chin Wang |
| collection | DOAJ |
| description | Abstract Hepatocellular carcinoma (HCC) is a highly invasive malignancy. Recently, GATOR1 (Gap Activity TOward Rags 1) complexes have been shown to play an important role in regulating tumor growth. NPRL2 is a critical component of the GATOR1 complex. Therefore, this study used NPRL2 knockdown to investigate how GATORC1 regulates the prognosis and development of HCC via the mammalian target of rapamycin (mTOR) and autophagy signaling pathways. We established HepG2 cells with NPRL2 knockdown using small interfering RNA (siRNA) and short hairpin RNA (shRNA) systems. The siRNA‐mediated and shRNA‐mediated NPRL2 down‐regulation significantly reduced the expression of NPRL2 and two other GATPOR1 complex components, NPRL3 and DEPDC5, in HepG2 cells; furthermore, the efficient down‐regulation of NPRL2 protein expression by both the shRNA and siRNA systems enhanced the proliferation, migration, and colony formation in vitro. Additionally, the NPRL2 down‐regulation significantly increased HCC growth in the subcutaneous and orthotopic xenograft mouse models. The NPRL2 down‐regulation increased the Rag GTPases and mTOR activation and inhibited autophagy in vitro and in vivo. Moreover, the NPRL2 level in the tumors was significantly associated with mortality, recurrence, the serum alpha fetoprotein level, the tumor size, the American Joint Committee on Cancer stage, and the Barcelona Clinic Liver Cancer stage. Low NPRL2, NPRL3, DEPDC5, and LC3, and high p62 and mTOR protein expression in the tumors was significantly associated with disease‐free survival and overall survival in 300 patients with HCC after surgical resection. Conclusion: The efficient down‐regulation of NPRL2 significantly increased HCC proliferation, migration, and colony formation in vitro, and increased HCC growth in vivo. Low NPRL2 protein expression in the tumors was closely correlated with poorer clinical outcomes in patients with HCC. These results provide a mechanistic understanding of HCC and aid the development of treatments for HCC. |
| first_indexed | 2024-03-12T18:47:34Z |
| format | Article |
| id | doaj.art-570acea9db984957a14221e692b0d8b3 |
| institution | Directory Open Access Journal |
| issn | 2471-254X |
| language | English |
| last_indexed | 2024-03-12T18:47:34Z |
| publishDate | 2022-12-01 |
| publisher | Wolters Kluwer Health/LWW |
| record_format | Article |
| series | Hepatology Communications |
| spelling | doaj.art-570acea9db984957a14221e692b0d8b32023-08-02T07:31:43ZengWolters Kluwer Health/LWWHepatology Communications2471-254X2022-12-016123563357710.1002/hep4.2019NPRL2 down‐regulation facilitates the growth of hepatocellular carcinoma via the mTOR pathway and autophagy suppressionYa‐Chin Wang0Ming‐Chao Tsai1Yaw‐Sen Chen2Pei‐Min Hsieh3Chao‐Ming Hung4Hung‐Yu Lin5Yao‐Chun Hsu6Jen‐Hao Yeh7Pojen Hsiao8Yu‐Cheih Su9Ching‐Hou Ma10Chih‐Yuan Lee11Chih‐Che Lin12Chih‐Wen Shu13Yu‐Chan Li14Mei‐Hsing Tsai15James Yu Lin16Wei‐Hao Peng17Ming‐Lung Yu18Chih‐Wen Lin19Division of Gastroenterology and Hepatology E‐Da Dachang Hospital I‐Shou University Kaohsiung TaiwanDivision of Hepato‐Gastroenterology Department of Medicine Kaohsiung Chang Gung Memorial Hospital Chang Gung University College of Medicine Kaohsiung TaiwanSchool of Medicine College of Medicine I‐Shou University Kaohsiung TaiwanSchool of Medicine College of Medicine I‐Shou University Kaohsiung TaiwanSchool of Medicine College of Medicine I‐Shou University Kaohsiung TaiwanSchool of Medicine College of Medicine I‐Shou University Kaohsiung TaiwanSchool of Medicine College of Medicine I‐Shou University Kaohsiung TaiwanDivision of Gastroenterology and Hepatology E‐Da Dachang Hospital I‐Shou University Kaohsiung TaiwanDivision of Gastroenterology and Hepatology E‐Da Dachang Hospital I‐Shou University Kaohsiung TaiwanSchool of Medicine College of Medicine I‐Shou University Kaohsiung TaiwanDepartment of Orthopedic Surgery E‐Da Hospital I‐Shou University Kaohsiung TaiwanDepartment of Surgery National Taiwan University Hospital Taipei TaiwanDepartment of Surgery Kaohsiung Chang Gung Memorial Hospital and Chang Gung University College of Medicine Kaohsiung TaiwanSchool of Medicine College of Medicine I‐Shou University Kaohsiung TaiwanDivision of Gastroenterology and Hepatology E‐Da Dachang Hospital I‐Shou University Kaohsiung TaiwanSchool of Medicine College of Medicine I‐Shou University Kaohsiung TaiwanDivision of Gastroenterology and Hepatology E‐Da Dachang Hospital I‐Shou University Kaohsiung TaiwanSchool of Medicine College of Medicine I‐Shou University Kaohsiung TaiwanHepatobiliary Section Department of Internal Medicine, Hepatitis Center Kaohsiung Medical University Hospital Kaohsiung TaiwanDivision of Gastroenterology and Hepatology E‐Da Dachang Hospital I‐Shou University Kaohsiung TaiwanAbstract Hepatocellular carcinoma (HCC) is a highly invasive malignancy. Recently, GATOR1 (Gap Activity TOward Rags 1) complexes have been shown to play an important role in regulating tumor growth. NPRL2 is a critical component of the GATOR1 complex. Therefore, this study used NPRL2 knockdown to investigate how GATORC1 regulates the prognosis and development of HCC via the mammalian target of rapamycin (mTOR) and autophagy signaling pathways. We established HepG2 cells with NPRL2 knockdown using small interfering RNA (siRNA) and short hairpin RNA (shRNA) systems. The siRNA‐mediated and shRNA‐mediated NPRL2 down‐regulation significantly reduced the expression of NPRL2 and two other GATPOR1 complex components, NPRL3 and DEPDC5, in HepG2 cells; furthermore, the efficient down‐regulation of NPRL2 protein expression by both the shRNA and siRNA systems enhanced the proliferation, migration, and colony formation in vitro. Additionally, the NPRL2 down‐regulation significantly increased HCC growth in the subcutaneous and orthotopic xenograft mouse models. The NPRL2 down‐regulation increased the Rag GTPases and mTOR activation and inhibited autophagy in vitro and in vivo. Moreover, the NPRL2 level in the tumors was significantly associated with mortality, recurrence, the serum alpha fetoprotein level, the tumor size, the American Joint Committee on Cancer stage, and the Barcelona Clinic Liver Cancer stage. Low NPRL2, NPRL3, DEPDC5, and LC3, and high p62 and mTOR protein expression in the tumors was significantly associated with disease‐free survival and overall survival in 300 patients with HCC after surgical resection. Conclusion: The efficient down‐regulation of NPRL2 significantly increased HCC proliferation, migration, and colony formation in vitro, and increased HCC growth in vivo. Low NPRL2 protein expression in the tumors was closely correlated with poorer clinical outcomes in patients with HCC. These results provide a mechanistic understanding of HCC and aid the development of treatments for HCC.https://doi.org/10.1002/hep4.2019 |
| spellingShingle | Ya‐Chin Wang Ming‐Chao Tsai Yaw‐Sen Chen Pei‐Min Hsieh Chao‐Ming Hung Hung‐Yu Lin Yao‐Chun Hsu Jen‐Hao Yeh Pojen Hsiao Yu‐Cheih Su Ching‐Hou Ma Chih‐Yuan Lee Chih‐Che Lin Chih‐Wen Shu Yu‐Chan Li Mei‐Hsing Tsai James Yu Lin Wei‐Hao Peng Ming‐Lung Yu Chih‐Wen Lin NPRL2 down‐regulation facilitates the growth of hepatocellular carcinoma via the mTOR pathway and autophagy suppression Hepatology Communications |
| title | NPRL2 down‐regulation facilitates the growth of hepatocellular carcinoma via the mTOR pathway and autophagy suppression |
| title_full | NPRL2 down‐regulation facilitates the growth of hepatocellular carcinoma via the mTOR pathway and autophagy suppression |
| title_fullStr | NPRL2 down‐regulation facilitates the growth of hepatocellular carcinoma via the mTOR pathway and autophagy suppression |
| title_full_unstemmed | NPRL2 down‐regulation facilitates the growth of hepatocellular carcinoma via the mTOR pathway and autophagy suppression |
| title_short | NPRL2 down‐regulation facilitates the growth of hepatocellular carcinoma via the mTOR pathway and autophagy suppression |
| title_sort | nprl2 down regulation facilitates the growth of hepatocellular carcinoma via the mtor pathway and autophagy suppression |
| url | https://doi.org/10.1002/hep4.2019 |
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