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目的:构建含有MDA-7基因的的真核表达载体并制备重组腺病毒,转染/感染肝癌细胞后,观察其对细胞增殖的影响。方法:构建MDA-7的真核表达载体,将表达质粒转染肝癌细胞,利用平板克隆形成实验观察MDA-7分子对肿瘤细胞系克隆形成能力的影响。利用Adeasy-1腺病毒重组系统构建、包装并扩增含有MDA-7基因的重组腺病毒。利用MTT实验检测Ad-MDA-7对肿瘤细胞增殖的影响。结果:成功构建了MDA-7真核表达载体,利用平板克隆形成实验证实其可抑制肝癌细胞的克隆形成能力。成功地构建并包装了含有MDA-7基因的重组腺病毒,利用MTT实验证明其可抑制肝癌细胞的增殖。结论:MDA-7对肝癌细胞的增殖具有显著的抑制作用,为进一步研究其在肝癌细胞中的功能提供了重要的实验依据。
OBJECTIVE: To construct a eukaryotic expression vector containing MDA-7 gene and prepare recombinant adenovirus. After transfection / infection of hepatoma cells, observe the effects on cell proliferation. Methods: The eukaryotic expression vector of MDA-7 was constructed. The expression plasmid was transfected into hepatoma cells. The effect of MDA-7 on the clonality of tumor cell lines was observed by plate clone formation assay. A recombinant adenovirus containing the MDA-7 gene was constructed, packaged and amplified using the Adeasy-1 adenovirus recombination system. The effect of Ad-MDA-7 on the proliferation of tumor cells was detected by MTT assay. Results: The eukaryotic expression vector of MDA-7 was successfully constructed and its clonogenic capacity of hepatocellular carcinoma cells was inhibited by plate clone formation assay. The recombinant adenovirus containing MDA-7 gene was successfully constructed and packaged, and MTT assay was used to prove that it can inhibit the proliferation of hepatoma cells. Conclusion: MDA-7 has a significant inhibitory effect on the proliferation of hepatoma cells, which provides an important experimental basis for further study of its function in hepatocellular carcinoma cells.