Scinderin基因沉默对人胃癌细胞SGC-7901增殖、转移的影响

Effects of Scinderin Silence on the Proliferation and Metastasis of Human Gastric Cancer Cell SGC-7901

Scinderin是一种依赖Ca2+的肌动蛋白丝(F-actin)切割蛋白,在细胞分泌过程中发挥着重要作用。目前,针对scinderin在人类疾病尤其是肿瘤中的生物学功能研究报道的并不多。该实验通过构建scinderin-shRNA慢病毒载体并感染人胃癌细胞株SGC-7901,于荧光显微镜下观测感染效率,利用RT-qPCR和Western blot实验证实scinderin的沉默效果。运用实时细胞分析仪(RTCA)检测细胞的增殖能力,流式细胞术检测细胞周期变化,Transwell小室检测细胞的迁移能力。结果显示,将构建好的病毒载体成功转入了胃癌细胞SGC-7901。感染scinderin-shRNA病毒载体后,scinderin的mRNA和蛋白质表达水平均受到不同程度的抑制(P<0.01),细胞的增殖和迁移能力均显著降低(P<0.05),细胞周期阻滞在G2/M期。该研究表明,胃癌细胞SGC-7901中scinderin低表达能有效抑制细胞的增殖和转移能力,这也为scinderin在胃癌演化过程中的机制研究奠定了实验基础。

Scinderin is a Ca2＋-dependent filamentous actin （F-actin） severing and capping protein, which plays a key role in secretion. But few researches about biological effects of scinderin on human disease especially neoplasm are currently reported. In this study, we transfected the shRNA targeting scinderin lentiviral vector into human gastric cancer cell line SGC-7901. The efficiency oftransfection was observed under fluorescence microscope. Effects of gene silencing were confirmed by RT-qPCR and Western blot. The proliferation was analyzed by real-time cell analyzer （RTCA） and the cell cycle distribution was investigated by flow cytometer. The migration was analyzed by Transwell. Results showed that scinderin-shRNA was transfected into SGC-7901 successfully. After transfection, levels of scinderin mRNA and protein expression reduced significantly （P〈0.01）. The abilities of cell proliferation and migration decreased obviously （P〈0.05）, while cell cycle was arrested in the G2/M phase. Consequently, we can draw the conclusion that silencing scinderin can effectively restrain the proliferation and metastasis of human gastric cancer cell SGC-7901, which will be an experiment support for the possible regulation of scinderin in gastric cancer evolvement.