慢病毒表达载体研究全长组织因子对乳腺癌的作用  

Study on role of tissue factor in breast cancer with lentivirus expression vector

作　　者：张艳杰[1,2] 武敏[1,2] 蒋贤杰 张渝君[1,2] 

机构地区：[1]四川大学生命科学学院,四川成都610064 [2]四川大学生长代谢衰老研究中心,四川成都610064

出　　处：《兰州大学学报（医学版）》2015年第3期1-6,12,共7页Journal of Lanzhou University（Medical Sciences）

基　　金：国家自然科学基金项目(31170729)

摘　　要：目的用慢病毒表达载体研究全长组织因子(flTF)在乳腺癌发生和发展中的作用。方法包装flTF的慢病毒颗粒感染永生化乳腺上皮细胞株MCF-10A实现flTF的过表达,用flTF sh RNA的慢病毒颗粒感染乳腺癌细胞株MDA-MB-231下调flTF的表达,用Transwell的方法检测flTF过表达和下调表达之后对细胞迁移的影响,用流式细胞术检测flTF对细胞周期的影响,蛋白印迹法检测flTF过表达和下调表达的效果,结合MTS方法检测flTF对细胞增殖的影响。结果 flTF在乳腺癌细胞株MDA-MB-231中的表达明显高于永生化的乳腺上皮细胞MCF-10A,正常细胞中flTF过表达促进了细胞的迁移并对其生长速度产生了一定的影响。在肿瘤细胞中下调flTF的表达,能在一定程度上抑制肿瘤细胞的迁移和生长。结论慢病毒表达载体在flTF的表达对乳腺癌细胞的影响中得到成功应用,flTF可作为抑制肿瘤细胞生长和迁移的靶分子,下调flTF的表达或许能为乳腺癌的临床治疗开辟一条新的途径。Objective To study the role of full-length tissue factor in initiation and development of breast cancer with lentivirus expression system. Methods Pack full-length tissue factor lentivirus particle and infect human non-tumorigenic immortalized breast epithelial cell line MCF-10A cells to overexpression full-length tissue factorin this cell line. Infect breast cancer cell line MDA-MB-231 with full-length tissue factor＇s shR-NA particles to knockdown full-length tissue factor. The effects of full-length tissue factor＇s overexpression and knock-down on cell migration were shown by Transwell assay, cell cycle was detected by Flow cytome-try, full-length tissue factor＇s overexpression and knockdown were detected by Western blot, the effect of tis-sue factor on cell proliferation was detected by MTS assay. Results The expression of full-length tissue factor in breast cancer cell line MDA-MB-231 was significantly higher than that in human non-tumorigenic immor-talized breast epithelial cell line MCF-10A cells , overexpression of full-length tissue factor in MCF-10A cells increased cell migration and growth, while knockdown of tissue factor in MDA-MB-231 cells, suppressed migration and proliferation of these malignant tumor cells. Conclusion These results suggest that Letivirus expression system was successfully used in the study of full-length tissue factor＇s role in breast cancer. Therefore, full-length tissue factor could be a target for treatment of breast cancer. Inhibition of full-length tissue factor expression would open up a novel approach to anti-breast cancer therapy.

关 键 词：全长组织因子 肿瘤细胞转移 慢病毒表达载体 RNA干扰 乳腺癌 

分 类 号：R737.9[医药卫生—肿瘤]