裂瘤型腺病毒Ad-E1A对小鼠腹水型肝癌的基因治疗及免疫功能影响

The Oncolytic Adenovirus Ad-E1A in the Gene Therapy of Mouse Ascitic Hepatoma in vivo and Its Effect on Immune Function

目的研究裂瘤型腺病毒Ad-E1A对BALB/c小鼠腹水型肝癌的基因治疗效果,并检测其对免疫功能的影响。方法将Ad-E1A进行多轮扩增获得高效价病毒子,采用RT-PCR法鉴定E1A基因在小鼠肝癌H22细胞内的转录。将BALB/c小鼠分为非致瘤组(正常组)和皮下注射H22细胞转移瘤的致瘤组,其中致瘤组又分为PBS组、Ad-E1A组、5-Fu组进行抗肿瘤试验,检测各组的抑瘤率、胸腺指数、脾脏指数、白细胞总数等。结果获得了达7×109pfu/ml的高滴度Ad-E1A。致瘤组的抗肿瘤试验中,Ad-E1A组与5-Fu组的抑瘤率分别为66.7%和68.8%,二者差异无统计学意义(P>0.05);Ad-E1A组和PBS组间的胸腺指数和脾脏指数变化无统计学意义,而5-Fu组显著降低(P<0.01和P<0.05);同时Ad-E1A组和PBS组间的白细胞总数变化不大,但5-Fu组显著降低(P<0.01),其中中性粒细胞、淋巴细胞、单核细胞均明显降低(P<0.05)。结论 Ad-E1A可以抑制小鼠腹水型肝癌的生长,且对小鼠的免疫功能未见明显毒副作用。

Objective To research the gene therapy of mouse ascitic hepatoma and its effect on immune function by oncolytic adenovirus Ad-E1A. Methods The E1A recombined adenovirus was proliferated in QBI-293A cells. RT-PCR was used to confirm the transcription of E1A gene in mouse hepatoma H22 cells. The tumor subcutaneous model was established with mouse ascitic hepatoma. The antitumor inhibitive ratio,thymus index,spleen index,and the number of white blood cell,lymphocyte,neutrophil and monocyte in every group were detected. Results The high titer E1A recombined adenovirus was obtained（ The titer is 7 × 109pfu/ml） . In tumor subcutaneous model,compared with PBS group,the inhibitive ratio of Ad-E1A and 5-Fu was 66. 7% and 68. 8% respectively（ P 0. 05） . Compared with invariable Ad-E1A group and PBS group,the thymus index and spleen index of 5-Fu were decreased obviously（ P 0. 01 P 0. 05） . The number of white blood cells were equivalence between Ad-E1A group and nontumor group,while the group of 5-Fu were drop rapidly（ P 0. 01） . The number of lymphocyte,neutrophil and monocyte were decreased obviously in 5-Fu group. Conclusion The oncolytic adenovirus AdE1A inhibite H22 ascitic hepatoma obviously,and almost has no side effect on immune function.