建立适用于旋毛虫抗肿瘤相关基因筛选的动物模型

Establishment of a Mouse Model Suitable for Screening of Trichinella spiralis Anti-Tumor Gene

目的为利用抑制消减杂交(SSH)法筛选旋毛虫抗肿瘤相关基因,建立理想的动物模型获取可靠的实验样本。方法Balb/c小鼠随机分为检测组(Tester)和驱动组(Driver),Tester为经口服感染旋毛虫250条、400条和550条组,Driver为不接种组,在感染后11 d,两组同时在皮下分别接种SP2/0细胞2×106个/只,检测组和驱动组小鼠荷瘤后,于20 d后处死,采集两组肿瘤组织和脾脏组织,用天平称量肿瘤块的重量,游标卡尺测量肿瘤块3个互相垂直直径进行体积比较;为了检测两组小鼠免疫情况,又采用流式细胞术检测体内T淋巴细胞的动态变化,以便评估所需样本的可靠性。结果将肿瘤组织与正常组织分离后,经统计分析比较两组肿瘤块重量和体积大小的差异均极为显著(P<0.01)通过检测T淋巴细胞亚类,FACS共检测1×105个细胞,分别得到脾细胞中CD3+、CD4+和CD8+T淋巴细胞的数量,感染组小鼠机体的CD3+、CD4+、CD8+和CD4+/CD8+显著增高,在接种不同剂量的旋毛虫后荷瘤,小鼠脾脏特异的T淋巴细胞亚类:CD3+差异显著(P<0.05);CD4+差异显著(P<0.05);CD8+差异显著(P<0.05);CD4+/CD8+差异显著(P<0.05)。通过以上指标的测定,我们认为所建立的动物模符合SSH的实验要求。结论建立的旋毛虫抗实体瘤动物模型,可用于旋毛虫抗肿瘤差异基因筛选的实验起始材料,为进一步研究旋毛虫抗肿瘤分子机制创造条件。

Objective In order to investigate the anti-tumor genes of Trichinella spiralis by suppression subtractive hybridization （SSH）, a suitable Balb/c mice anti-tumor model was established and the anti-tumor effects of Trichinella spiralis infection was studied. Methods 6 - 8-week old Balb/c male mice were randomized into tester and driver groups. For the tester group,the mice were infected orally with 250,400 and 550 T. spiralis larvae. The mice of driver group were not treated with T. spiralis larvae. At the llth day after infection,the mice were inoculated subcutaneously with 2 ×10^6 SP^2/0 cells. The mice were killed at the 20^th day after tumor cell inoculation. The size and weight of tumor were measured. The T lymphocyte diversity was detected by flow cytometry. Statistical differences between the two groups were determined by Student＇s test. Results The tumors were isolated, and the tumor volume and weight were measured and analyzed. Significant difference was found between the two groups （ P 〈 0.01 ）. The anti-tumor effect of Trichinella spiralis was also observed by determining the number of CD 3 ＋ ,CD4 ＋ ,CD8 ＋ and CIM ＋/CD8 ＋ lymphocytes of the mice. There were more CD3 ＋ , CD4 ＋ , CD8 ＋ , and CD4 ＋/CD8 ＋ cells in the tester group than that of driver group （ P 〈 0.05）. The samples from animal model were suitable for SSH experiment. Conclusion A Trichinella spiralis infected mouse model of anti-tumor effects has been established, which is suitable for experiment for differentially expression gene screening. The mouse model may lay a foundation for further studies on the mechanism of TrichineUa spiralis anti-tumor effects.