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目的 :探讨逆转录病毒载体介导的单纯疱疹病毒胸苷激酶 (HSV1 tk)基因转染 ,联合抗病毒药物羟甲基无环鸟苷 ( ganciclovir ,GCV)和无环鸟苷 (acyclovir ,ACV )对人卵巢上皮癌 (OEC)细胞系TYK的杀伤作用。方法 :用lipofectionreagent介导法将PLNTK5质粒转移入包装细胞PA317中 ,用滴度最高的PA317病毒上清液转染TYK细胞 ,运用MTT法及光镜、电镜检测GCV、ACV对目的细胞的体外杀伤效应。结果 :将PLNTK 5质粒转入PA317细胞后 ,得到了稳定的产病毒细胞株。TYK细胞可被病毒上清液转染 ,转导HSV1 tk基因可显著增强ACV、GCV对TYK细胞的杀伤作用。结论 :逆转录病毒可介导HSV1 tk基因转染TYK细胞并获稳定表达 ,ACV、GCV对携有HSV1 tk基因的卵巢癌细胞有明显的体外杀伤作用。
Objective: To investigate retroviral vector-mediated HSV1 tk gene transfection, combined with antiviral drugs ganciclovir (GCV) and acyclovir (ACV) Cytotoxicity on human ovarian epithelial carcinoma (OEC) cell line TYK. METHODS: PLNTK5 plasmid was transfected into PA317 packaging cells by lipofectionreagent mediated method. TYK cells were transfected with PA317 virus supernatant with the highest titer. MTT assay and light microscopy were used to detect the cytotoxicity of GCV and ACV on the target cells in vitro effect. Results: After transfecting the PLNTK 5 plasmid into PA317 cells, a stable virus-producing cell line was obtained. TYK cells can be transfected with virus supernatant, transduction of HSV1 tk gene can significantly enhance the ACV, GCV TYK cell killing. CONCLUSION: The retrovirus can transduce HSV1 tk gene into TYK cells stably and express in vitro. ACV and GCV have obvious cytotoxicity on ovarian cancer cells carrying HSV1 tk gene in vitro.