HSV-TK/GCV系统在难治性卵巢癌裸鼠移植瘤模型中的实验研究

目的:建立人难治性卵巢癌荷瘤裸鼠模型,用HSV-TK/GCV自杀基因系统治疗模型动物,观察疗效及协同因素。方法:常规培养携带HSV-TK基因的重组逆转录病毒包装细胞PA317,测定病毒滴度。在移植瘤内多点注射包装细胞,观察GCV、ATRA、TPT对移植肿瘤独立及协同治疗后的肿瘤体积变化。治疗结束后分析移植肿瘤组织病理,PCR检测TK基因在NIH3T3细胞及移植肿瘤组织的转导。结果:人卵巢癌标本裸鼠原代移植成功率16.7%,鼠间传代移植成功率89.7%;重组逆转录病毒滴度为6×104cfu/ml;HSV-TK/GCV系统或TPT治疗后肿瘤生长受到抑制(P<0.05),两者联合治疗后,抑制更明显(P<0.05);ATRA灌胃未显示独立或协同治疗作用(P>0.05);HSV-TK/GCV组、TPT组及两者联合组有明显的治疗反应;PCR检测证实,TK基因在NIH3T3细胞及肿瘤组织中的转导。结论:HSV-TK/GCV系统及TPT对人难治性卵巢癌荷瘤裸鼠有单独及协同治疗作用,全反式维甲酸灌胃对人难治性卵巢癌荷瘤裸鼠疗效不明显。

Study of HSV-TK/GCV system in the nude mice xenograft tumor model of human refractory ovarian carcinoma

Objective:To establish a model of xenograft tumor of human refractory ovarian carcinoma in nude mice and to observe the independent and cooperative curative effects of HSV-TK/GCV system,TPT and ATRA in this model.Methods:Retroviral vector containing HSV-TK gene was transfected into PA317 packaging cell,then these PA317 cells were cultivated conventionally.Retroviral vectors were then titrated.VPCs were injected at multiple sites of xenograft tumors.After independent and cooperative therapy of HSV-TK/GCV,ATRA and TPT,the volume of tumors were observed,then we analysed the pathologic changes of the tumor tissue and used PCR to detect TK gene in the NIH3T3 cell and the xenograft tumors.Results:One of six times(16.7%),nude mice transplanted with human refractory ovarian carcinoma tissue developed subcutaneous tumor.Total achievement ratio of tumor transplantation between the nude mice was 89.7%.The titer of retroviral was 6×10~4.HSV-TK/GCV system and TPT not only restrained the growth of xenograft tumors independently (P<0.05),but also cooperated each other(P<0.05).There was obviously pathologic differences of tumor tissue between the groups mentioned above and their comparisons.PCR confirmed that TK gene had been transferred into the NIH3T3 cell the xenograft tumor.ATRA administered by oral gavage hadn' t restrained the growth of xenograft tumors,furthermore,it wasn' t cooperative with HSV-TK/GCV system and TPT(P>0.05).Conclusions:There were independent and cooperative effects of HSV-TK/GCV system and TPT;ATRA administered by oral gavage had no obvious effect in this trial.