SPARC基因过表达对卵巢癌淋巴结高转移细胞生物学特性的影响

发布时间：2018-03-20 04:12

本文选题：卵巢肿瘤　切入点：淋巴结转移　出处：《中国细胞生物学学报》2015年03期 　论文类型：期刊论文

【摘要】：该研究旨在探讨富含半胱氨酸的酸性分泌蛋白基因(secreted protein acidic and rich in cysteine gene,SPARC)过表达对卵巢癌淋巴结高转移细胞(SKOV3-PM4)生物学特性的影响。构建SPARC基因的慢病毒表达载体并转染SKOV3-PM4细胞,Real-time PCR和Western blot验证转染后的表达效率,激光共聚焦免疫荧光进行蛋白的细胞定位,细胞计数法和集落形成实验测定细胞增殖能力,流式细胞仪检测细胞周期,Transwell小室实验测定细胞体外侵袭、迁移能力。实验结果显示,SPARC蛋白存在于核周及胞质;过表达SPARC基因后,SKOV3-PM4细胞增殖受到明显抑制(P0.05);细胞周期检测结果显示,各期改变无明显差异;体外侵袭、迁移实验结果显示,SKOV3-PM4细胞侵袭、迁移能力显著降低(P0.05)。实验结果表明,SPARC基因在卵巢癌淋巴结转移中可能发挥抑癌基因的生物学作用。
[Abstract]:The aim of this study was to investigate the effect of overexpression of cysteine-rich protein acidic and rich in cysteine gene Sparc on the biological characteristics of ovarian cancer cell line SKOV3-PM4. Construction of SPARC gene expression vector combined with lentivirus expression. Real-time PCR and Western blot were used to verify the expression efficiency of transfected SKOV3-PM4 cells. Laser confocal immunofluorescence was used to locate the protein, cell count and colony formation assay were used to determine the cell proliferation ability, and flow cytometry was used to detect cell cycle and cell invasion in vitro. Migration ability. The results showed that the SPARC protein existed in the nucleus and cytoplasm, the proliferation of SKOV3-PM4 cells was significantly inhibited after overexpression of SPARC gene, and the cell cycle test showed that there was no significant difference in the changes of cell cycle and invasion in vitro. The results of migration assay showed that SKOV3-PM4 cells invaded and the migration ability of SKOV3-PM4 cells decreased significantly. The results showed that SPARC gene may play a biological role as a tumor suppressor gene in lymph node metastasis of ovarian cancer.
【作者单位】：广西医科大学附属肿瘤医院;广西医科大学医学科学实验中心;
【基金】：国家自然基金(批准号:81060218;81360502) 广西自然科学基金(批准号:2014GXNSFAA118161)资助的课题~~
【分类号】：R737.31

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