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目的观察单纯疱疹病毒-胸苷激酶/胞嘧啶脱氨酶融合自杀基因(HSV-TK/CD)对骨肉瘤细胞增殖的影响。方法将含TK/CD的重组腺病毒载体感染人骨肉瘤细胞系MG-63细胞,测定感染效率后,分别给予不同浓度的前体药物和不同感染复数的自杀基因重组腺病毒,计算瘤细胞增殖抑制率,MTT法测定细胞存活率,并用流式细胞术和Hochest33342染色法分析杀伤机制。结果含自杀基因的重组腺病毒成功感染MG-63细胞;不同感染复数和不同前体药物浓度下,融合自杀基因系统对瘤细胞的杀伤作用明显强于单基因和双基因相加的作用(P<0.05);其杀伤机制为致细胞坏死和细胞凋亡。结论融合自杀基因HSV-TK/CD对骨肉瘤细胞系MG-63细胞的生长有明显的抑制作用。
Objective To observe the effect of herpes simplex virus-thymidine kinase / cytosine deaminase fusion suicide gene (HSV-TK / CD) on the proliferation of osteosarcoma cells. Methods The recombinant adenoviral vector containing TK / CD was transfected into human osteosarcoma cell line MG-63. After being assayed for infection efficiency, different doses of prodrug and recombinant suicide gene adenovirus with different multiplicity of infection were respectively administered to calculate the tumor cell proliferation inhibition The cell viability was determined by MTT assay and the killing mechanism was analyzed by flow cytometry and Hochest33342 staining. Results The recombinant adenovirus containing suicide gene successfully infected MG-63 cells. The killing effect of fusion suicide gene system on tumor cells was significantly stronger than that of single gene and double gene under different concentrations and different prodrug concentrations (P <0.05). The mechanism of killing was necrosis and apoptosis. Conclusion The fusion suicide gene HSV-TK / CD significantly inhibits the growth of osteosarcoma cell line MG-63.