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目的研究nm23-H1基因转染对人舌癌Tca8113细胞株生物学行为的影响。方法采用脂质体转染法将真核表达载体pAdEasy-nm23-H1转染Tca8113细胞,分为Tca8113-Ad-nm23-H1组(转染pAdEasy-nm23-H1)、Tca8113-Ad组(转染空载体pAdEasy)和Tca8113组(未转染)。采用RT-PCR检测nm23-H1基因在细胞中的表达,噻唑蓝比色法检测转染前、后细胞体外增殖能力的变化,流式细胞术分析转染前、后细胞周期及凋亡的改变,采用Transwell小室和冲刷实验观察细胞侵袭、黏附、趋化运动能力的变化,克隆形成实验观察转染前、后细胞克隆形成能力。结果与Tca8113-Ad组和Tca8113组比较,Tca8113-Ad-nm23-H1组细胞体外增殖能力、细胞侵袭、黏附、趋化运动能力、克隆形成率及细胞S期比例下降(P<0.05);细胞G2/M期比例及凋亡率明显增高(P<0.05)。结论 nm23-H1基因转染后对人舌癌Tca8113细胞的增殖能力、侵袭转移能力均有明显抑制作用;nm23-H1基因可明显抑制人舌癌Tca8113细胞G2期向M期过渡,同时促进癌细胞凋亡。
Objective To investigate the effect of nm23-H1 gene transfection on the biological behavior of human tongue cancer Tca8113 cell line. Methods The eukaryotic expression vector pAdEasy-nm23-H1 was transfected into Tca8113 cells by lipofection method and divided into Tca8113-Ad-nm23-H1 group (transfected with pAdEasy-nm23-H1) and Tca8113- Empty vector pAdEasy) and Tca8113 group (untransfected). The expression of nm23-H1 gene in cells was detected by RT-PCR. The proliferation of cells was detected by MTT assay before and after transfection. The changes of cell cycle and apoptosis were analyzed by flow cytometry Transwell chambers and scouring experiments were used to observe the changes of cell invasion, adhesion and chemotaxis motility. Clonal formation assay was used to observe the cell clone formation ability before and after transfection. Results Compared with Tca8113-Ad group and Tca8113 group, the cell proliferation, cell invasion, adhesion, chemotaxis, colony formation rate and the proportion of S phase in Tca8113-Ad-nm23-H1 group decreased G2 / M phase ratio and apoptosis rate was significantly increased (P <0.05). Conclusion nm23-H1 gene transfection can significantly inhibit the proliferation, invasion and metastasis of human tongue cancer Tca8113 cells. Nm23-H1 gene can significantly inhibit the transition from G2 phase to M phase of human tongue cancer Tca8113 cells and promote the proliferation of cancer cells Apoptosis.