旋毛虫感染诱导的小鼠乳腺癌MCF-7细胞基因表达的变化

目的构建旋毛虫感染诱导的小鼠乳腺癌MCF-7细胞基因表达变化的cDNA消减文库,探讨旋毛虫感染诱导的MCF-7细胞基因表达的变化。方法复制MCF-7细胞小鼠模型,设对照组和旋毛虫感染组(实验组),采用抑制性消减杂交技术构建旋毛虫感染诱导的MCF-7细胞上调基因表达消减文库,并利用反向Northernblot技术对所获得基因的特异性表达进行验证。结果成功构建了旋毛虫感染诱导的MCF-7细胞上调基因表达消减文库,克隆的目的基因片段大小分布在200～500bp之间,20个阳性克隆的测序结果经BLAST比对,获得差异表达的已知功能基因8个和未知功能基因4个,这些基因在小鼠乳腺癌细胞中均有表达,而在肌肉组织中无表达。结论旋毛虫感染诱导的MCF-7细胞差异表达基因上调,其中RB基因可能与乳腺癌细胞的生长抑制密切相关,为在基因水平研究旋毛虫抗肿瘤机理奠定了基础。

Change of Gene Expression in Breast Cancer MCF-7 Cells Induced by Trichinella spiralis Infection in Mice

Objective To construct a cDNA substractive library for investigation on Trichinella spiralis infection-induced change of gene expression in breast cancer MCF-7 cells. Methods MCF-7 cell-bearing model mice were replicated and divided into control and test groups randomly. Each mouse in test group was fed with 400 Trichinella spiralis 11 d after inoculation with MCF-7 cells, while that in control group with physiological saline. All the mice were killed 20 d after inoculation with MCF-7 cells, and their tumor tissues were isolated for construction of gene expression up-regulation subtractive library of Trichinella spiralis infection-induced MCF-7 cells by suppression subtraction hybridization (SSH)technique. The specific expressions of obtained genes were verified by reverse Northern blot. Results The gene expression up-regulation subtractive library of Trichinella spiralis infection-induced MCF-7 cells was successfully constructed. The lengths of cloned target gene fragments ranged from 200 to 500 bp. By analysis of sequencing results of 20 positive clones using BLAST software, 12 differentially expressed genes, 8 known and 4 unknown, were obtained, of which RB was a cancer suppressor gene. All the genes were expressed in breast cells but not in muscle tissue of mice. Conclusion The expression of differential gene including cancer suppressor gene RB in Trichinella spiralis infection-induced MCF-7 cells was upregulated, which laid a foundation of study on anti-tumor mechanism of Trichinella spiralis at gene level.