腺病毒介导HSV-TK/GCV与IL-2基因对人涎腺腺样囊性癌的抑制作用

目的探讨腺病毒介导的单纯疱疹病毒胸苷激酶（Herpes simplex virus thymidine kinase/ganciclovir,HSV-TK/GCV）与白细胞介素-2（interleukin-2,IL-2）基因联合应用对涎腺腺样囊性癌的抑制和杀伤作用。方法将0.2 ml细胞浓度为2×107/ml的涎腺腺样囊性癌高转移细胞株（salivary adenoid cystic carcinoma cell line withhigh tendency of lung metastasis,ACC-M）的细胞悬液接种于20只裸鼠皮下,建立涎腺腺样囊性癌裸鼠移植瘤模型。随机将20只荷瘤裸鼠分为A组：HSV-TK/GCV和IL-2联合基因治疗组（TK（IL-2组）;B组：HSV-TK/GCV组（TK组）;C组：空病毒组（EGFP组）;D组：ACC-M组;每组5只。分别于瘤体内注射TK（IL-2;TK;EGFP和无菌生理盐水。注射48小时后,对A、B、C组裸鼠每日腹腔注射GCV,D组每日腹腔注射生理盐水,连续10天。最后一次给药后第二天处死裸鼠,观察各组裸鼠的肿瘤体积和瘤重的变化,计算肿瘤生长抑制率;对移植瘤进行组织病理学观察;应用流式细胞仪检测移植瘤的细胞凋亡情况。结果在A组和B组中,ACC-M的生长均受到显著抑制。治疗后体积数减少,与ACC-M组比较差别有极显著性（P〈0.01）。A组和B组抑瘤率分别为91%和76%;光镜下观察A组和B组均出现了明显的细胞凋亡;流式细胞仪检测A组和B组有明显的凋亡峰出现,凋亡率分别为30.55±0.23和13.53±0.32;C组凋亡率为10.07±0.31,D组凋亡率为8.34±0.41%。除C、D组之间差别无明显性外,其余各组之间差别均有极显著性（P〈0.01）。A组和B组的细胞S期比例明显增加,G2/M期比例明显降低。结论 HSV-TK/GCV联合IL-2基因对ACC-M移植瘤有明显抑制作用,抑瘤效果优于单独应用HSV-TK/GCV系统。

Inhibiting effect of combined HSV-TK/GCV with IL-2 gene therapy by adenovirus vector on salivary adenoid cystic carcinoma

Objective To investigate the inhibiting effect of combined Herpes simplex virus thymidine kinase/ganciclovir（HSV-TK/GCV） with interleukin-2（IL-2） gene transfected by adenovirus vector on salivary adenoid cystic carcinoma（SACC）.Methods SACC cell line with high tendency of lung metastasis（ACC-M） was used in this study.A concentration of 2×107 permilliliter of 0.2 ml ACC-M cell liquids was injected 20 nude mice subcutaneously to establish the animal model.The animals were divided randomly into group A（group TK＋IL-2）,group B（group TK）,group C（group EGFP） and group D（group ACC-M） of 5 mice each.HSV-TK and IL-2,HSV-TK,EGFP and normal saline was injected the nude mice of group A,B,C,and D respectively.Forty-eight hours after injection,100mg GCV/kg was injected nude mice of group A、B、C,twice daily for 10 days.Group D was injected normal saline as the control.The following day after the last medicine was given,all nude mice were killed.The weight and volume of tumor was measured respectively.The ratio of tumor inhibition was calculated.The histological changes of transplanted tumor of animal model were observed by light microscope.The percentage of apoptotic cells was detected by flow cytometer.Results After gene treatment,the growth of the transplanted tumor of nude mice was significantly inhibited,and the tumor volumes of group A and B were decreased（P0.01）.The ratio of the tumor inhibition of group A was 91%,group B was 76%.The appearent apoptosis of transplanted tumor was observed by light microscope.The results of flow cytometer showed that a typical apoptotic peak was formed in group A and B respectively.The percentage of apoptotic cells was 30.55±0.23% in groupA,13.53±0.32% in group B,10.07±0.31% in group C and 8.34±0.41% in group D.There was significantly difference between either two groups among the four groups（P0.01）,except between group C and group D.The cells in S phase and G2/M phase of both group A and B were significantly increased and decreased respectively than those of group C and group D（P0.01）.Conclusion HSV-TK/GCV and IL-2 could suppress the growth of ACC-M transplanted tumors of nude mice.The effect of combined HSV-TK with IL-2 on ACC-M transplanted tumor was much better than that of HSV-TK used alone.