| 摘要: |
| 目的观察人类新血小板反应素R-spondin 3(Rspo 3)的细胞定位，并探讨其在肿瘤发生发展中的可能作用。方法利用荧光显微成像法观察EGFP-Rspo 3在HEK293细胞中的分布。将重组质粒pcDNA-Rspo 3转染结肠癌细胞HT-29、LoVo，采用流式细胞术观察Rspo 3基因过表达对肿瘤细胞周期与凋亡的影响；采用Matrigel、Transwell实验分别检测Rspo 3基因过表达对肿瘤细胞黏附及侵袭能力的影响。结果荧光显微成像法观察发现，EGFP-Rspo 3融合蛋白在细胞核表达，呈弥散点状分布。流式细胞术观察发现，Rspo 3基因过表达对肿瘤细胞生长周期没有明显影响；Rspo 3基因过表达不影响低恶性的HT-29细胞凋亡，但诱导高恶性的LoVo细胞凋亡(P<0.01)。Rspo 3基因过表达增强肿瘤细胞黏附性(P<0.01)，降低肿瘤细胞的侵袭力(P<0.01)，对肿瘤细胞移行有一定抑制作用。结论Rspo 3有核定位功能，能诱导部分肿瘤细胞凋亡并抑制其转移扩散。 |
| 关键词: R-spondin 3 亚细胞定位 细胞周期 细胞凋亡 细胞黏附 肿瘤侵润 |
| DOI：10.3724/SP.J.1008.2011.0249 |
| 投稿时间：2010-12-24修订日期：2011-02-13 |
| 基金项目:上海市科委自然科学基金(044119655). |
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| Cellular localization of a novel human thrombospondin R-spondin 3 and its effects on tumor cells |
| YANG Wei-zhi1, 4， SUN Qing-li2， CHEN Jin-zhong3， HU Zhi-qian4* |
| (1. Department of Orthopedics, Tenth People’s Hospital of Shanghai, Tongji University, Shanghai 200072, China; 2. Department of Research and Development, MicroPort Medical (Shanghai) Co., Ltd, Shanghai 201203, China; 3. Department of Gene Therapy, School of Life Sciences, Fudan University,Shanghai 200433, China; 4. Department of General Surgery, Changzheng Hospital, Second Military Medical University, Shanghai 200003, China) |
| Abstract: |
| ObjectiveTo study the cellular localization of a novel human thrombospondin R-spondin 3 and its role in the development and progression of tumors.MethodsThe subcellular localization of R-spondin 3 was investigated in HEK293 cells by fluorescence micrography. Colon carcinoma cell lines HT-29 and LoVo were transfected with pcDNA-Rspo 3 recombinant expression plasmid; and the cell cycle and apoptosis were detected by Flow Cytometry (FCM). Cell adhesion and invasion ability were determined by Matrigel reagents and Transwell system, respectively.ResultsFluorescence micrography showed that EGFP-Rspo 3 fusion protein was mainly localized in nuclei as dispersed particles. Overexpression of R-spondin 3 showed no effect on cell cycle in both colon carcinoma cell lines. Overexpression of R-spondin 3 induced apoptosis of LoVo cells with high malignancy (P<0.01) although it showed no effect on the apoptosis of HT-29 cells with low malignancy. The overexpression also promoted the adhesion ability (P<0.01), restrained invasion ability (P<0.01) and motility of both colon carcinoma cell lines.ConclusionOur research indicates that R-spondin 3 localize in the nuclei; it can induce apoptosis and restrain metastatic potential of some tumor cells. |
| Key words: R-spondin 3 subcellular location cell cycle apoptosis cell adhesion neoplasm invasiveness |
