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目的:研究转染nm23-H1基因对体外培养A549细胞增殖和侵袭的抑制作用及其机理。方法:构建nm23-H1基因的真核表达载体,转染到体外培养的人肺腺癌细胞A549中,通过G418筛选出稳定表达克隆,RT-PCR及免疫组化检测nm23-H1在细胞内的表达情况。绘制细胞生长曲线检测nm23-H1基因对细胞生长的影响,流式细胞仪检测细胞周期,原子力显微镜观察细胞膜表面伪足的超微结构。结果:转染后的肿瘤细胞能稳定表达nm23-H1基因,抑制了肿瘤细胞的增殖。nm23-H1基因没有诱导细胞凋亡但使G1期细胞增加而S期细胞减少,停滞于G0期。转染nm23-H1基因后细胞边缘的伪足减少。结论:nm23-H1基因能抑制体外培养的A549肿瘤细胞的增殖,可能通过改变细胞表面结构减弱细胞的侵袭能力。
Objective: To investigate the inhibitory effect and its mechanism of nm23-H1 gene transfection on proliferation and invasion of A549 cells in vitro. Methods: The eukaryotic expression vector of nm23-H1 gene was constructed and transfected into human lung adenocarcinoma A549 cells in vitro. Stable clones were screened by G418. RT-PCR and immunohistochemistry were used to detect the expression of nm23-H1 Express the situation. The cell growth curve was used to detect the effect of nm23-H1 gene on the cell growth. The cell cycle was detected by flow cytometry. The ultrastructure of pseudopodia on the cell membrane was observed by atomic force microscopy. Results: The transfected tumor cells stably expressed nm23-H1 gene and inhibited the proliferation of tumor cells. The nm23-H1 gene did not induce apoptosis but increased the number of G1 phase cells and the S phase cells, arresting in G0 phase. Pseudopodia decreased at the cell periphery after transfection of the nm23-H1 gene. Conclusion: The nm23-H1 gene can inhibit the proliferation of A549 tumor cells cultured in vitro, which may reduce the invasion ability of cells by changing the cell surface structure.