| 唐秋琳,郎 楠,李黎博,石 芳,毕 锋.鸟苷酸交换因子P92GEF通过靶向调控RhoA抑制肿瘤细胞增殖侵袭（英文稿）[J].现代生物医学进展英文版,2019,19(16):3018-3027. |
| 鸟苷酸交换因子P92GEF通过靶向调控RhoA抑制肿瘤细胞增殖侵袭（英文稿） |
| Guanine Exchange Factor P92GEF target RhoA and Inhibit Cell Proliferation and Invasion as a Tumor gene |
| Received:December 07, 2018 Revised:December 31, 2018 |
| DOI：10.13241/j.cnki.pmb.2019.16.004 |
| 中文关键词: 鸟苷酸交换因子 P92GEF Dbl家族 Rho家族 癌基因 |
| 英文关键词: Guanine nucleotide exchange factors P92GEF Dbl family Rho GTPases Tumor gene |
| 基金项目:国家自然科学基金项目（30701719；30971519） |
| Author Name | Affiliation | E-mail | | TANG Qiu-lin | Department of laboratory molecular targeted therapy of tumor and medical oncology, West China Hospital,
Sichuan University, Chengdu, Sichuan, 610041, China | tangqiulin1981@163.com | | LANG Nan | Department of laboratory molecular targeted therapy of tumor and medical oncology, West China Hospital,
Sichuan University, Chengdu, Sichuan, 610041, China | | | LI Li-bo | Department of laboratory molecular targeted therapy of tumor and medical oncology, West China Hospital,
Sichuan University, Chengdu, Sichuan, 610041, China | | | SHI Fang | Department of laboratory molecular targeted therapy of tumor and medical oncology, West China Hospital,
Sichuan University, Chengdu, Sichuan, 610041, China | | | BI Feng | Department of laboratory molecular targeted therapy of tumor and medical oncology, West China Hospital,
Sichuan University, Chengdu, Sichuan, 610041, China | |
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| 中文摘要: |
| 摘要：Dbl家族鸟苷交换因子（GEFs）是Rho家族蛋白发生恶性转化的主要调控单位，它通过使Rho蛋白从无活性的GDP形式转换为GTP形式的Rho蛋白而发挥作用，参与细胞骨架重排，细胞的生长和活力。P92GEF是一GEFs家族分子。本研究通过Real time PCR对P92GEF在人体48种正常组织中的表达情况进行了测定；GST-pulldown技术对P92GEF的体内GEF活性进行了检测；双荧光素酶报告基因检测技术对下游小分子进行转录因子活性检测；应用免疫荧光双染标记法完成了高表达P92GEF对正常细胞骨架形态的影响；在细胞表型实验中分别使用CCK8法、Transwell法及软琼脂克隆形成实验检测了高表达P92GEF对细胞增殖侵袭迁移及体外成瘤能力的影响。研究结果显示P92GEF有841个氨基酸，具有典型的Dbl家族分子结构域，在肺组织中表达量最高，能够促使正常成纤维细胞中的应力纤维（stress fiber）增多，P92GEF转染的NIH3T3细胞可以独立生长和形成继发性病灶，同时促使细胞增殖，侵袭及克隆形成能力增强，体外转录因子活性检测发现该基因可能与JAK/STAT通路有关。因此，P92GEF是一个典型的鸟苷交换因子家族分子，能激活Rho家族分子RhoA，具有明显的癌基因特征。 |
| 英文摘要: |
| ABSTRACT: Dbl family guanine exchange factor (GEFs) is the main regulatory unit for the malignant transformation of Rho family proteins. It plays an important role in the transformation of Rho protein from the inactive GDP form to the active GTP form and participates in the cytoskeleton rearrangement, the cell growth and vitality. This study conducted a preliminary structural and functional study of a typical family member, P92GEF, and discussed the role of the molecule in the development of tumor. The expression of P92GEF in 48 normal tissues of human body was measured by Realtime PCR; GST-pulldown technique was used to detect the GEF activity of P92GEF in vivo; the dual-luciferase reporter gene detection technique was used to test the transcription factor activity of the small downstream molecules, and the high expression of P92GEF was tested by the double staining immunofluorescence method. Effects of high expression of P92GEF on cell proliferation, invasion and tumorigenic ability in vitro were examined using CCK8, Transwell and soft agar clony formation assays. The results of bioinformatics analysis showed that P92GEF contains 841 amino acids, has typical Dbl family molecular domain. The expression of P92GEF in the lung tissue was highest. P92GEF binds to RhoA in vitro and promotes the proliferation and invasion of NIH3T3 cells, and has significant ability to clone in vitro. P92GEF is a typical guanine exchange factor family molecule, which can activate Rho family RhoA and has obvious oncogene characteristics. |
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