摘要: |
目的: 体外制备高迁移率组蛋白N2(high mobility group chromosal protein N2,HMGN2)蛋白,观察其能否转运入骨肉瘤细胞及入胞后的细胞定位。方法: 构建HMGN2过表达慢病毒质粒并包装慢病毒,在HEK-293FT细胞中过表达并纯化。用纯化的HMGN2蛋白处理骨肉瘤细胞。通过免疫细胞化学检测外源性HMGN2能否转运入骨肉瘤细胞及细胞定位。结果: 成功制备、纯化HMGN2蛋白,并发现外源性HMGN2能转运入骨肉瘤细胞,主要定位于细胞核。结论: 外源性HMGN2能转运入细胞并定位于细胞核,为其发挥生物学效应奠定基础。 |
关键词: 骨肉瘤 转移 高迁移率组蛋白N2 蛋白纯化 |
DOI:10.12025/j.issn.1008-6358.2020.20191521 |
分类号:R738.1 |
基金项目:国家自然科学基金(81772305,U1603118),上海市科学技术委员会科研计划(16140901800). |
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Study on the entry of recombinant HMGN2 protein into osteosarcoma cells and cell localization |
MA Xiao, XU En-jie, YIN Jia, ZHOU Xu-hui
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Department of Spine Surgery, Changzheng Hospital, Naval Medical University, Shanghai 200003, China
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Abstract: |
Objective: To prepare high mobility group chromosal protein N2 (HMGN2) protein in vitro, and observe its capacity to enter into osteosarcoma cells and its cell localizations. Methods: After packaging HMGN2 overexpressing lentivirus, the recombinant lentivirus was delivered into HEK-293FT cells. HMGN2 protein was purified from the harvested 293FT cells using protein purification kit. Purified HMGN2 protein was used to treat cultured osteosarcoma cells. Whether the exogenous HMGN2 could enter into osteosarcoma cells and its cell localizations were detected by immunocytochemistry assay. Results: HMGN2 protein was successfully prepared and purified. The exogenous HMGN2 protein entered into osteosarcoma cells in culture, and mainly located in the nucleus. Conclusions: HMGN2 could be successfully prepared and purified, and can enter into cells. It mainly localizes in the nucleus, to lay a foundation for its biological effects. |
Key words: osteosarcoma metastasis high mobility group chromosal protein N2 protein purification |