目的：探讨LASP⁃1在人舌鳞状细胞癌细胞中的表达，并应用shRNA沉默人舌鳞状细胞癌细胞SCC9中LASP⁃1基因，研究LASP⁃1对SCC9细胞增殖和迁移能力的影响。方法构建携带绿色荧光蛋白的LASP⁃1 shRNA质粒及阴性对照组质粒LASP⁃1 shNC，通过脂质体转染SCC9细胞，采用MTT法检测细胞增殖；RT⁃PCR法检测细胞LASP⁃1 mRNA表达；Western blot检测细胞LASP⁃1蛋白的表达；运用Transwell小室实验检测细胞迁移能力。结果 LASP⁃1在舌鳞状细胞癌细胞中高表达；实验组和阴性对照组分别转染相应质粒48 h后细胞均有绿色荧光蛋白表达，表明质粒转染成功；实验组SCC9细胞LASP⁃1 mRNA和LASP⁃1蛋白表达明显降低；与空白对照组相比，实验组细胞培养48 h和72 h细胞存活率分别降低了（51.23±1.47）%和（50.07±2.11）%；Transwell实验结果显示， LASP⁃1基因被沉默后细胞迁移能力明显下降，比对照组降低约43%。结论 LASP⁃1在舌鳞状细胞癌细胞中高表达，下调LASP⁃1基因表达可抑制SCC9细胞的增殖和迁移。%Objective To investigate the expression of LASP⁃1 in tongue squamous cell carcinoma, and to study the influence of shRNA silent LASP⁃1 on cell proliferation and migration of tongue squamous cell carcinoma. Methods Green fluorescence shRNA targeting LASP⁃1 was constructed and transfected into SCC9 cells by lipofectamine. Cell proliferation was detected by MTT method. LASP⁃1 mRNA and protein were determined by RT⁃PCR and western blot respectively. The migration ability of cells was detected by transwell assay. Results There was green fluorescence ex⁃pression in experimental and negative control group cells after transfecting shRNA and shNC respectively. LASP⁃1 mRNA and protein were decrease in experimental group cells. It indicated that LASP⁃1 shRNA was transfected success⁃fully. By contrast with control group, the cell viability of experimental group cells reduced by (51.23 ± 1.47)% and (50.07 ± 2.11)% after 48 h and 72 h respectively. The results of transwell assay showed that migration ability of SCC9 cells was decreased significantly after shRNA targeting LASP⁃1 was successful transfected. It decreased by 43% com⁃pared with control group. Conclusion LASP⁃1 has high expression in tongue squamous cell carcinoma cells, down reg⁃ulated LASP⁃1 gene expression can inhibit the proliferation and migration of SCC9 cells.
展开▼
