Cullinl基因对乳腺癌细胞迁移和侵袭的影响

The role of Cullinl in breast cancer cells migration and invasion

目的研究Cullinl（Cull）基因对乳腺癌细胞迁移及侵袭的影响，并探讨其具体分子作用机制。方法体外化学合成CullsiRNA和ControlsiRNA（si-Ctrl）序列，转染乳腺癌MDA—MB-231和BT-549细胞，细胞迁移和侵袭实验观察抑制Cull的表达对2种乳腺癌细胞迁移和侵袭能力的影响；明胶酶谱实验检测Cull对2种乳腺癌细胞分泌基质金属蛋白酶（MMPs）的影响；Westernblot实验观察Cull对2种乳腺癌细胞中金属蛋白酶组织抑制因子（TIMPs）及MMPs蛋白表达的影响。结果抑制Cull的表达可以明显降低2种乳腺癌细胞的迁移和侵袭能力；Cull的表达降低可以抑制2种乳腺癌细胞分泌MMP-2；抑制Cull的表达可以通过上调TIMP-2的蛋白表达而抑制MMP-2的蛋白表达。结论抑制Cull的表达可以通过上调TIMP-2的表达从而抑制MMP-2的表达，打破了TIMP-2／MMP-2的平衡，最终抑制了乳腺癌细胞的迁移和侵袭能力。

Objective To investigate the role of Cullinl （ Cull ） gene in cell migration, invasion and its molecular mechanisms in MDA- MB -231 and BT- 549 breast cancer cell fines. Methods Chemically synthesized Cull siRNA （ si - Cull ） and Control siRNA （ si - Ctrl） were transfected into human breast cancer cell lines MDA - MB - 231 and BT -549 by siLentFect Lipid Reagent. The migration and invasion abilities of MDA - MB -231 and BT -549 cells after Cull silencing were detected by cell migration and invasion assay. We performed gelatin zymography to measure the MMPs activities in breast cancer cells and Western blot to examine the TIMPs and MMPs proteins expressions after Cull knockdown. Results Cull knockdown decreased cell migration and invasion abilities of MDA - MB -231 and BT -549 cells. The MMP - 2 activity was significantly suppressed after knockdown of Cull. Silencing Cull unregulated TIMP - 2 protein expression and inhibited MMP - 2 protein expression. Conclusion Knockdown of Cull can result in the up - regulation of TIMP - 2, down - regulation of MMP - 2 and the imbalance between TIMP - 2 and MMP - 2 greatly contrib- utes to the reduced cell migration and invasion abilities in breast cancer cells.