The RXFP2-PLC/PKC signaling pathway mediates INSL3-induced regulation of the proliferation, migration and apoptosis of mouse gubernacular cells
Abstract Background Testicular hypoplasia can affect the sexual and reproductive ability in adulthood, and even increase the risk of cancer. Abnormal development of the gubernaculum is one of the important factors of testicular hypoplasia. Therefore, a study of the structure and function of the gube...
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| Format: | Article |
| Language: | English |
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BMC
2023-02-01
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| Series: | Cellular & Molecular Biology Letters |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s11658-023-00433-0 |
| _version_ | 1827983337661136896 |
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| author | Shouxing Duan Xuewu Jiang Jianhong Li Maxian Fu Zhuo Li Yiyi Cheng Yangmu Zhuang Ming Yang Wenfeng Xiao Hongyan Ping Yao Xie Xiaojun Xie Xuan Zhang |
| author_facet | Shouxing Duan Xuewu Jiang Jianhong Li Maxian Fu Zhuo Li Yiyi Cheng Yangmu Zhuang Ming Yang Wenfeng Xiao Hongyan Ping Yao Xie Xiaojun Xie Xuan Zhang |
| author_sort | Shouxing Duan |
| collection | DOAJ |
| description | Abstract Background Testicular hypoplasia can affect the sexual and reproductive ability in adulthood, and even increase the risk of cancer. Abnormal development of the gubernaculum is one of the important factors of testicular hypoplasia. Therefore, a study of the structure and function of the gubernaculum is an important but neglected new breakthrough point for investigating the normal/abnormal development of the testis. Previous findings showed that Insulin like factor 3 (INSL3) is a key factor regulating the growth of gubernaculum, however, the mechanism by which INSL3 acts on the gubernaculum remains unknown. Therefore, we probed the mechanism associated with INSL3-induced the proliferation, migration, and apoptosis of gubernacular cells in mice. Methods A culture cell model of neonatal mice gubernaculum is established by INSL3 intervention. We blocked PLC/PKC signaling pathway with U73122 pretreat to investigate the role of the PLC/PKC signaling pathway. The changes of cell proliferation, migration, and apoptosis were detected by molecular biological methods. In addition, the levels of PCNA and F-action were detected by immunofluorescence and western blotting. Results We found that INSL3 can promote the proliferation and migration of gubernacular cells and inhibit their apoptosis, meanwhile, INSL3 significantly up-regulated PLC/PKC protein phosphorylation. However, treatment with the PLC/PKC signaling pathway inhibitor U73122 significantly inhibited these effects of INSL3. Besides, we found that INSL3 could up-regulate the protein expression level of PCNA and F-actin, while the PCNA and F-actin expression was significantly weakened after U73122 pretreatment. Conclusions This research revealed that INSL3 binding to RXFP2 may up-regulate the expression levels of PCNA and F-actin by activating the PLC/PKC signaling pathway to promote the proliferation and migration of gubernacular cells. It suggests that the RXFP2-PLC/PKC axis may serve as a novel molecular mechanism by which INSL3 regulates growth of the gubernaculum. |
| first_indexed | 2024-04-09T22:46:00Z |
| format | Article |
| id | doaj.art-7fc8dddc4f6744c398e18baf91818dcd |
| institution | Directory Open Access Journal |
| issn | 1689-1392 |
| language | English |
| last_indexed | 2024-04-09T22:46:00Z |
| publishDate | 2023-02-01 |
| publisher | BMC |
| record_format | Article |
| series | Cellular & Molecular Biology Letters |
| spelling | doaj.art-7fc8dddc4f6744c398e18baf91818dcd2023-03-22T11:49:57ZengBMCCellular & Molecular Biology Letters1689-13922023-02-0128111410.1186/s11658-023-00433-0The RXFP2-PLC/PKC signaling pathway mediates INSL3-induced regulation of the proliferation, migration and apoptosis of mouse gubernacular cellsShouxing Duan0Xuewu Jiang1Jianhong Li2Maxian Fu3Zhuo Li4Yiyi Cheng5Yangmu Zhuang6Ming Yang7Wenfeng Xiao8Hongyan Ping9Yao Xie10Xiaojun Xie11Xuan Zhang12Department of Pediatric Surgery, Huazhong University of Science and Technology Union Shenzhen Hospital (Nanshan Hospital)Department of Pediatric Surgery, Pingshan District Maternal and Child Healthcare Hospital of Shenzhen, Pingshan General Hospital of Southern Medical UniversityDepartment of Pediatric Surgery, The Second Affiliated Hospital of Shantou University Medical CollegeDepartment of Pediatric Surgery, Huazhong University of Science and Technology Union Shenzhen Hospital (Nanshan Hospital)Department of Pediatric Surgery, Huazhong University of Science and Technology Union Shenzhen Hospital (Nanshan Hospital)Department of Pediatric Surgery, The First Affiliated Hospital of Shantou University Medical CollegeDepartment of Pediatric Surgery, The First Affiliated Hospital of Shantou University Medical CollegeDepartment of Pediatric Surgery, The First Affiliated Hospital of Shantou University Medical CollegeDepartment of Pediatric Surgery, The Second Affiliated Hospital of Shantou University Medical CollegeDepartment of Pediatric Surgery, Pingshan District Maternal and Child Healthcare Hospital of Shenzhen, Pingshan General Hospital of Southern Medical UniversityDepartment of Radiology, Cancer Hospital of Shantou University Medical CollegeDepartment of General Surgery, The First Affiliated Hospital of Shantou University Medical CollegeDepartment of Pediatric Surgery, Pingshan District Maternal and Child Healthcare Hospital of Shenzhen, Pingshan General Hospital of Southern Medical UniversityAbstract Background Testicular hypoplasia can affect the sexual and reproductive ability in adulthood, and even increase the risk of cancer. Abnormal development of the gubernaculum is one of the important factors of testicular hypoplasia. Therefore, a study of the structure and function of the gubernaculum is an important but neglected new breakthrough point for investigating the normal/abnormal development of the testis. Previous findings showed that Insulin like factor 3 (INSL3) is a key factor regulating the growth of gubernaculum, however, the mechanism by which INSL3 acts on the gubernaculum remains unknown. Therefore, we probed the mechanism associated with INSL3-induced the proliferation, migration, and apoptosis of gubernacular cells in mice. Methods A culture cell model of neonatal mice gubernaculum is established by INSL3 intervention. We blocked PLC/PKC signaling pathway with U73122 pretreat to investigate the role of the PLC/PKC signaling pathway. The changes of cell proliferation, migration, and apoptosis were detected by molecular biological methods. In addition, the levels of PCNA and F-action were detected by immunofluorescence and western blotting. Results We found that INSL3 can promote the proliferation and migration of gubernacular cells and inhibit their apoptosis, meanwhile, INSL3 significantly up-regulated PLC/PKC protein phosphorylation. However, treatment with the PLC/PKC signaling pathway inhibitor U73122 significantly inhibited these effects of INSL3. Besides, we found that INSL3 could up-regulate the protein expression level of PCNA and F-actin, while the PCNA and F-actin expression was significantly weakened after U73122 pretreatment. Conclusions This research revealed that INSL3 binding to RXFP2 may up-regulate the expression levels of PCNA and F-actin by activating the PLC/PKC signaling pathway to promote the proliferation and migration of gubernacular cells. It suggests that the RXFP2-PLC/PKC axis may serve as a novel molecular mechanism by which INSL3 regulates growth of the gubernaculum.https://doi.org/10.1186/s11658-023-00433-0INSL3RXFP2PLC/PKCGubernaculumSignaling pathway |
| spellingShingle | Shouxing Duan Xuewu Jiang Jianhong Li Maxian Fu Zhuo Li Yiyi Cheng Yangmu Zhuang Ming Yang Wenfeng Xiao Hongyan Ping Yao Xie Xiaojun Xie Xuan Zhang The RXFP2-PLC/PKC signaling pathway mediates INSL3-induced regulation of the proliferation, migration and apoptosis of mouse gubernacular cells Cellular & Molecular Biology Letters INSL3 RXFP2 PLC/PKC Gubernaculum Signaling pathway |
| title | The RXFP2-PLC/PKC signaling pathway mediates INSL3-induced regulation of the proliferation, migration and apoptosis of mouse gubernacular cells |
| title_full | The RXFP2-PLC/PKC signaling pathway mediates INSL3-induced regulation of the proliferation, migration and apoptosis of mouse gubernacular cells |
| title_fullStr | The RXFP2-PLC/PKC signaling pathway mediates INSL3-induced regulation of the proliferation, migration and apoptosis of mouse gubernacular cells |
| title_full_unstemmed | The RXFP2-PLC/PKC signaling pathway mediates INSL3-induced regulation of the proliferation, migration and apoptosis of mouse gubernacular cells |
| title_short | The RXFP2-PLC/PKC signaling pathway mediates INSL3-induced regulation of the proliferation, migration and apoptosis of mouse gubernacular cells |
| title_sort | rxfp2 plc pkc signaling pathway mediates insl3 induced regulation of the proliferation migration and apoptosis of mouse gubernacular cells |
| topic | INSL3 RXFP2 PLC/PKC Gubernaculum Signaling pathway |
| url | https://doi.org/10.1186/s11658-023-00433-0 |
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