pLXSN-HPVL2-Angiostatin的构建及转染T细胞淋巴瘤细胞系的体外杀伤活性研究

Study of the Construction of pLXSN-HPVL2-Angiostatin and Its Cytolysis Capacity on Transfected TLymphoma Hut-78 Cell in Vitro

目的 探讨人乳头瘤病毒 (HPV)感染相关疾病既具靶向性又能阻断局部血管生长的基因治疗新途径。方法 将血管生长抑制因子 (angiostatin)及HPV抗原基因同时克隆入逆转录病毒表达载体pLXSN ,用PA3 17细胞包装、筛选 ,建立高滴度的感染性重组病毒产生细胞系 ,以重组病毒转染T细胞淋巴瘤Hut 78细胞系 ,在体外观察其对HPV感染阳性宫颈癌细胞的杀伤活性。结果 PCR及RT PCR检测显示获得了 pLXSN HPVL2 Angiostatin ,并成功转染Hut 78细胞系 ,镜下观察及MTT检测 ,显示了其对HPV阳性宫颈癌细胞的特异趋向及高杀伤活性。结论 pLXSN HPVL2 An giostatin及其成功转染的Hut

Objective To lay a solid foundation for both t he treatment and the research of HPV related diseases,we probe a new method whic h can not only target HPV related cells but also block up angiogenin.Methods First angiostatin and HPVL2 cDNA were cloned into pLXSN at the same time and the recombined pLXSN-HPVL2-Angiostatin was packed by PA317 cells using cellfectin, then a high titer cell line that produced recom bined pLXSN-HPVL2-Angiostatin was selected.After that T lymphoma Hut-78 cell s were transfected by pLXSN-HPVL2-Angiostatin using Polybrene. Last the cytoly sis capacity of transfected T lymphoma Hut-78 cell was observed by microscope a nd MTT in vitro.Results PCR and RT-PCR revealed that both the recombined pLXSN-HPVL2-Angiostatin and its transfected T lymphoma Hut-78 cell were acqu ired.The observation revealed the targeting and cytolysis capacity of transfecte d T lymphoma Hut-78 cell to HPV related tumor cells in vitro.Conclusion The recombined pLXSN-HPVL2-Angiostatin and i ts transfected T lymphoma Hut-78 cell provide a new way to genetherapy of HPV r elated diseases.