| Residential College | false |
| Status | 已發表Published |
| EBP50 influences distribution and localization of microfilament cytoskeleton in cultured HeLa cells | |
| Zheng J.2; Wang Y.2; Chen P.2; He J.2 | |
| 2008-12-01 | |
| Source Publication | Chinese Journal of Clinical Oncology
 |
| ISSN | 16727118 |
| Volume | 35Issue:20Pages:1192-1195 |
| Abstract | Objective: To explore the influence of EBP50 (ezrin-radixin-moesin-binding phospho-protein-50) on microfilament cytoskeleton content and distribution in cultured HeLa cells, to investigate the relationship between the changes in microfilament cytoskeleton localization and EBP50 after PDGF (platelet-derived growth factor) stimulation, and to further clarify the molecular mechanism by which EBP50 suppresses tumor cell proliferation and migration. Methods: pBK-CMV-HA-EBP50 wild type recombinant plasmid and pBK-CMV-HA empty vector were transfected into HeLa cells. G418 at 350mg/L was used to screen for cell clones stably expressing EBP50. Western blot was carried out to detect EBP50 expression. Similarities and differences in microfilament cytoskeleton content and distribution in HeLa cells transfected with pBK-CMV-HA-EBP50 wild type recombinant plasmid or pBK-CMV-HA empty vector were analyzed by Western blot, fluorescence staining and confocal microscopy. HeLa cells stably transfected with the pBK-CMV-HA-EBP50 wild type recombinant plasmid and the pBK-CMV-HA empty vector were also treated with PDGF (10 ng/ml and 20 ng/ml, 37°C, 15min) and stained by rhodamine-labeled phalloidin to observe the distribution of microfilament cytoskeleton in the two groups. EBP50 protein distribution in PDGF-stimulated HeLa cells was detected by immunofluorescence. Results: Western blot results confirmed that the EBP50 cDNA fragment could express EBP50 in cultured HeLa cell lines and that cell lines stably expressing EBP50 were successfully obtained. Western blot and fluorescence results showed that in the cell line transfected with empty vector, the microfilament cytoskeleton was thick, loose, multidirectional and displayed crossing arrangements. The content of microfilament cytoskeleton in the cell line transfected with pBK-CMV-HA-EBP50 was different from that found in the cell line transfected with empty vector. EBP50 expression enhanced microfilament cytoskeleton polymerization into compact thin filaments. Under the stimulation of PDGF, EBP50 migrated to the cell membrane from the cytosol together with microfilament cytoskeleton and co-localized there. Conclusion: EBP50 can change the distribution of microfilament cytoskeleton in cultured HeLa cells and can also bind the microfilament cytoskeleton to the cell membrane under the stimulation of PDGF. EBP50 may play a role in the proliferation and migration of tumor cells by influencing the distribution and localization of microfilament cytoskeleton. |
| Keyword | EBP50 HeLa cell Microfilament cytoskeleton |
| URL | View the original |
| Language | 英語English |
| Fulltext Access | |
| Document Type | Journal article |
| Collection | University of Macau |
| Affiliation | 1.Shandong Eye Institute and Hospital 2.Capital Medical University China |
| Recommended Citation GB/T 7714 | Zheng J.,Wang Y.,Chen P.,et al. EBP50 influences distribution and localization of microfilament cytoskeleton in cultured HeLa cells[J]. Chinese Journal of Clinical Oncology, 2008, 35(20), 1192-1195. |
| APA | Zheng J.., Wang Y.., Chen P.., & He J. (2008). EBP50 influences distribution and localization of microfilament cytoskeleton in cultured HeLa cells. Chinese Journal of Clinical Oncology, 35(20), 1192-1195. |
| MLA | Zheng J.,et al."EBP50 influences distribution and localization of microfilament cytoskeleton in cultured HeLa cells".Chinese Journal of Clinical Oncology 35.20(2008):1192-1195. |
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