Effects of RhoC downregulation on the angiogenesis characteristics of myeloma vascular endothelial cells
Abstract Background Tumor angiogenesis plays an important role in disease progression, and RhoC has been previously found to be expressed in vascular endothelial cells (VECs); however, its role in tumor angiogenesis requires clarification. This study aimed to explore the effects of RhoC downregulati...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Wiley
2019-07-01
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| Series: | Cancer Medicine |
| Subjects: | |
| Online Access: | https://doi.org/10.1002/cam4.2208 |
| _version_ | 1818613191857405952 |
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| author | Zhihua Zhao Kai Liu Xiangyu Tian Miaomiao Sun Na Wei Xiaoyan Zhu Hongmei Yang Tong Wang Guozhong Jiang Kuisheng Chen |
| author_facet | Zhihua Zhao Kai Liu Xiangyu Tian Miaomiao Sun Na Wei Xiaoyan Zhu Hongmei Yang Tong Wang Guozhong Jiang Kuisheng Chen |
| author_sort | Zhihua Zhao |
| collection | DOAJ |
| description | Abstract Background Tumor angiogenesis plays an important role in disease progression, and RhoC has been previously found to be expressed in vascular endothelial cells (VECs); however, its role in tumor angiogenesis requires clarification. This study aimed to explore the effects of RhoC downregulation on the cytoskeleton, pseudopod formation, migration ability, and canalization capacity of myeloma vascular endothelial cells (MVECs) in vitro. Materials and methods The expression of RhoC in MVECs and human umbilical vein endothelial cells (HUVECs) was knocked down by shRNA, and the expression levels of RhoC mRNA were detected by quantitative reverse transcription polymerase chain reaction (qRT‐PCR). The cytoskeletal changes and pseudopods were observed by laser scanning confocal and scanning electron microscopy; VECs were incubated in two‐dimensional Matrigel and three‐dimensional microcarriers to observe tube‐like structures and budding status, respectively. The protein expression of RhoC, phosphorylation of mitogen‐activated protein kinase (p‐MAPK), and Rho‐associated coiled‐coil kinase (ROCK) was determined by Western blotting. The expression of RhoC in VECs was downregulated by RhoC shRNA, thereby decreasing the number of pseudopods, two‐dimensional tube‐like structures, and buds. Results When RhoC was downregulated, the expression levels of ROCK and phosphorylation of MAPK were both decreased (P < 0.05). Moreover, the expression levels of RhoC and phosphorylation of MAPK and three‐dimensional budding numbers were higher in MVECs than in HUVECs (P < 0.05). The downregulation of RhoC expression in MVECs and HUVECs inhibited pseudopod formation, migration, canalization ability, and angiogenesis (P < 0.05). Conclusion Our data indicated that MVECs and HUVECs were well suited for angiogenesis research, but the former cell type was shown to be more advantageous in terms of budding numbers. RhoC plays a pivotal role in MVECs angiogenesis, and the downregulation of RhoC expression could inhibit angiogenesis via the RhoC/MAPK and RhoC/ROCK signaling pathways. |
| first_indexed | 2024-12-16T15:58:12Z |
| format | Article |
| id | doaj.art-6ebd4751e178462eb15d2d59ff5933ac |
| institution | Directory Open Access Journal |
| issn | 2045-7634 |
| language | English |
| last_indexed | 2024-12-16T15:58:12Z |
| publishDate | 2019-07-01 |
| publisher | Wiley |
| record_format | Article |
| series | Cancer Medicine |
| spelling | doaj.art-6ebd4751e178462eb15d2d59ff5933ac2022-12-21T22:25:32ZengWileyCancer Medicine2045-76342019-07-01873502351010.1002/cam4.2208Effects of RhoC downregulation on the angiogenesis characteristics of myeloma vascular endothelial cellsZhihua Zhao0Kai Liu1Xiangyu Tian2Miaomiao Sun3Na Wei4Xiaoyan Zhu5Hongmei Yang6Tong Wang7Guozhong Jiang8Kuisheng Chen9Henan Province Key Laboratory of Tumor Pathology Department of Pathology of The First Affiliated Hospital of Zhengzhou University Zhengzhou People’s Republic of ChinaZhengzhou Central Hospital Affiliated to Zhengzhou University Zhengzhou Henan People’s Republic of ChinaDepartment of Pathology of The First Affiliated Hospital of Zhengzhou University Zhengzhou Henan People’s Republic of ChinaThe Affiliated Tumor Hospital of Zhengzhou University Zhengzhou Henan People’s Republic of ChinaHenan Province Key Laboratory of Tumor Pathology Department of Pathology of The First Affiliated Hospital of Zhengzhou University Zhengzhou People’s Republic of ChinaDepartment of Histology and Embryology, School of Basic Medical Sciences Zhengzhou University Zhengzhou Henan People’s Republic of ChinaHenan Province Medical College Zhengzhou Henan People’s Republic of ChinaDepartment of Pathology of The First Affiliated Hospital of Zhengzhou University Zhengzhou Henan People’s Republic of ChinaHenan Province Key Laboratory of Tumor Pathology Department of Pathology of The First Affiliated Hospital of Zhengzhou University Zhengzhou People’s Republic of ChinaHenan Province Key Laboratory of Tumor Pathology Department of Pathology of The First Affiliated Hospital of Zhengzhou University Zhengzhou People’s Republic of ChinaAbstract Background Tumor angiogenesis plays an important role in disease progression, and RhoC has been previously found to be expressed in vascular endothelial cells (VECs); however, its role in tumor angiogenesis requires clarification. This study aimed to explore the effects of RhoC downregulation on the cytoskeleton, pseudopod formation, migration ability, and canalization capacity of myeloma vascular endothelial cells (MVECs) in vitro. Materials and methods The expression of RhoC in MVECs and human umbilical vein endothelial cells (HUVECs) was knocked down by shRNA, and the expression levels of RhoC mRNA were detected by quantitative reverse transcription polymerase chain reaction (qRT‐PCR). The cytoskeletal changes and pseudopods were observed by laser scanning confocal and scanning electron microscopy; VECs were incubated in two‐dimensional Matrigel and three‐dimensional microcarriers to observe tube‐like structures and budding status, respectively. The protein expression of RhoC, phosphorylation of mitogen‐activated protein kinase (p‐MAPK), and Rho‐associated coiled‐coil kinase (ROCK) was determined by Western blotting. The expression of RhoC in VECs was downregulated by RhoC shRNA, thereby decreasing the number of pseudopods, two‐dimensional tube‐like structures, and buds. Results When RhoC was downregulated, the expression levels of ROCK and phosphorylation of MAPK were both decreased (P < 0.05). Moreover, the expression levels of RhoC and phosphorylation of MAPK and three‐dimensional budding numbers were higher in MVECs than in HUVECs (P < 0.05). The downregulation of RhoC expression in MVECs and HUVECs inhibited pseudopod formation, migration, canalization ability, and angiogenesis (P < 0.05). Conclusion Our data indicated that MVECs and HUVECs were well suited for angiogenesis research, but the former cell type was shown to be more advantageous in terms of budding numbers. RhoC plays a pivotal role in MVECs angiogenesis, and the downregulation of RhoC expression could inhibit angiogenesis via the RhoC/MAPK and RhoC/ROCK signaling pathways.https://doi.org/10.1002/cam4.2208angiogenesislentiviral vectormyelomaRhoCvascular endothelial cells |
| spellingShingle | Zhihua Zhao Kai Liu Xiangyu Tian Miaomiao Sun Na Wei Xiaoyan Zhu Hongmei Yang Tong Wang Guozhong Jiang Kuisheng Chen Effects of RhoC downregulation on the angiogenesis characteristics of myeloma vascular endothelial cells Cancer Medicine angiogenesis lentiviral vector myeloma RhoC vascular endothelial cells |
| title | Effects of RhoC downregulation on the angiogenesis characteristics of myeloma vascular endothelial cells |
| title_full | Effects of RhoC downregulation on the angiogenesis characteristics of myeloma vascular endothelial cells |
| title_fullStr | Effects of RhoC downregulation on the angiogenesis characteristics of myeloma vascular endothelial cells |
| title_full_unstemmed | Effects of RhoC downregulation on the angiogenesis characteristics of myeloma vascular endothelial cells |
| title_short | Effects of RhoC downregulation on the angiogenesis characteristics of myeloma vascular endothelial cells |
| title_sort | effects of rhoc downregulation on the angiogenesis characteristics of myeloma vascular endothelial cells |
| topic | angiogenesis lentiviral vector myeloma RhoC vascular endothelial cells |
| url | https://doi.org/10.1002/cam4.2208 |
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