INI1诱导上皮样肉瘤细胞向脂肪方向分化  

INI1 Induces Adipocytic Differentiation of Epithelioid Sarcoma Cells

作　　者：谢显彪[1,2] 温丽丽 吕东明[1,2] 邹雨桐 姚浩 彭挺生 XIE Xian-biao;WEN Li-li;LV Dong-ming;ZOU Yu-tong;YAO Hao;PENG Ting-sheng(Department of Musculoskeletal Oncology,The First Affiliated Hospital of Sun Yat-sen University,Guangzhou 510080,China;Guangdong Provincial Key Laboratory of Orthopedics and Traumatology,Guangzhou 510080,China;Department of Anesthesiology,Sun Yat-sen University Cancer Center,Guangzhou 510060,China;Department of Pathology,The First Affiliated Hospital of Sun Yat-sen University,Guangzhou 510080,China)

机构地区：[1]中山大学附属第一医院骨肿瘤科,广东广州510080 [2]广东省骨科学重点实验室,广东广州510080 [3]中山大学肿瘤防治中心麻醉科,广东广州510060 [4]中山大学附属第一医院病理科,广东广州510080

出　　处：《中山大学学报（医学科学版）》2020年第5期690-696,共7页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：国家自然科学基金(81202118);中山大学青年教师培育项目(19ykpy198)。

摘　　要：【目的】本研究旨在探讨INI1在上皮样肉瘤细胞中作用及机制。【方法】利用Western blot检测并比较上皮样肉瘤细胞中INI1表达水平;免疫组化检测上皮样肉瘤组织标本中INI1的表达;利用Tet-on系统重建上皮样肉瘤细胞VA-ES-BJ中INI1表达;诱导VA-ES-BJ细胞中INI1表达后检测细胞形态、增殖、脂肪相关分子标志,进一步Western blot检测脂肪分化相关转录因子表达及油红染色检测向脂肪方向分化的程度。【结果】在上皮样肉瘤细胞和临床标本中均存在INI1蛋白表达缺失的现象(P<0.05)。通过Tet-on系统在VA-ES-BJ细胞中成功重建INI1表达,经过强力霉素诱导后出现INI1表达上调。持续诱导INI1表达,VA-ES-BJ细胞形态发生改变,胞浆出现大量空泡;细胞增殖速度明显抑制(P<0.05);检测上皮来源细胞标记物,发现Cytokeratin明显下调(P<0.05);检测脂肪细胞相关标志物,发现瘦蛋白、脂联素、脂蛋白脂酶表达明显上调(P<0.05);进一步检测诱导INI1表达后脂肪分化相关转录因子PPAR-γ和CEBP-α表达均明显上调(P<0.05),油红染色呈显著阳性。【结论】上皮样肉瘤中存在INI1蛋白表达缺失现象,重建INI1表达通过上调转录因子PPAR-γ和CEBP-α表达,进而诱导上皮样肉瘤细胞向脂肪方向分化。【Objective】To study the function and mechanism of INI1 in human epithelioid sarcoma cells.【Methods】Western blotting was used to test the expressionlevel of INI1in epithelioid sarcoma cells.The expression of INI1 in clinical specimens of epithelioid sarcoma was detected by immunohistochemistry.Tet-on system was used to establish the expres⁃sion of INI1 in epithelioid sarcoma cells.Following the induction of INI1 expression,the cell morphology,proliferation and the molecular markers of adipocytic differentiation were detected.Then western blot was employed to detect the tran⁃scription factors of adipocytic differentiation,and oil red O staining was also tested.【Results】The expression of INI1 was absent in both epithelioid sarcoma cells and clinical tissues(P<0.05).The INI1 expression in the epithelioid sarcoma cells was successfully established by Tet-on system.After induction by doxycycline,the expression of INI1 was up-regu⁃lated.With continuous expression of INI1,the morphology of VA-ES-BJ cells was changed,with the appearance of abun⁃dant vacuoles in the cytoplasm.The cell proliferation was obviously inhibited(P<0.05).Epithelial cell marker Cytokera⁃tin was found significantly reduced(P<0.05).In addition,the adipocyte markers,leptin,adiponectin and lipoprotein lipase were significantly up-regulated(P<0.05).The adipogenic transcriptional factors PPAR-γand CEBP-αwere found upregulated(P<0.05)and the oil red staining was significantly positive.【Conclusions】The expression of INI1 was absent in epithelioid sarcoma.Reconstruction of the expression of INI1 could induce adipocytic differentiation via upregulation of transcriptional factors PPAR-γand CEBP-αin epithelioid sarcoma cells.

关 键 词：整合酶相互作用分子1 上皮样肉瘤 分化 

分 类 号：R738[医药卫生—肿瘤]