人信号素基因semaphorin-3F瞬时转染对舌鳞癌细胞增殖作用的影响

目的探讨人信号素家族中最具代表性成员信号素3F（semaphorin-3F，SEMA-3F）基因瞬时转染对舌鳞状细胞癌细胞生长的影响。方法将Tea8113细胞分为4组，即转染组：转染pEGFP-N1-SEMA-3F-cDNA的Tca8113细胞；空载体组：转染pEGFP-N1质粒的Tca8113细胞；未转染组：无转染细胞；对照组：仅加转染试剂。构建和纯化SEMA-3F的真核表达载体pEGFP-N1-SEMA-3F，用阳离子脂质体转染试剂介导转染Tca8113细胞。观察外源目的基因在靶细胞中的表达及细胞的生长情况。结果基因转染后48h成功检测到高表达SEMA-3F基因。转染后24、48、72、96h，甲基噻唑基四唑（methyl thiazolyl tetrazolium，MTT）法检测转染组的各时期平均A值，均低于其余3组。转染后48h，流式细胞仪检测G1期细胞比例降低，S期比例增高。结论SEMA-3F基因瞬时转染可使其在Tca8113细胞中高表达，并使细胞增殖受到抑制。本实验结果将SEMA-3F作为目的基因，为治疗舌癌进一步提供了实验依据。

Effect of human semaphoring-3F gene transient transfection on the proliferation of tongue carcinoma cells

OBJECTIVE: To investigate the effect of semaphorin-3F (SEMA-3F) gene transient transfection on the proliferation of Tca8113 tongue carcinoma cells. METHODS: After construction of a full-length SEMA3F expression vector, Tca8113 cells were transient transfected with pEGFP-N1-SEMA3F by Lipofectamine 2000. The expression of SEMA-3F gene was detected by RT-PCR The differences of the expression in the transfected cell groups, empty vector groups and untransfected cell groups were compared. The survival rates were assayed by methyl thiazolyl tetrazolium (MTT) enzymatic labeling technique. Cell cycle were assayed by flow cytometer (FCM). RESULTS: The gene was transfected into Tca8113 cells. High expression of SEMA-3F was successfully detected in the transfected cell groups after gene transfection. The cell cycle percentage of G1 stage decreased and S stage increased. CONCLUSIONS: SEMA-3F gene transient transfection may inhibit the proliferation of Tca8113 cells.