摘要
目的:观察Erbin基因表达沉默后对乳腺癌细胞形态及侵袭迁移生物学行为的影响。方法:通过慢病毒shRNA转染技术建立低表达Erbin的乳腺癌细胞模型。蛋白质印迹法检测基因沉默前后Erbin蛋白表达的效果;倒置显微镜观察基因沉默前细胞形态改变;Matrigel-Transwell小室法检测细胞侵袭能力改变;Transwell小室法检测细胞迁移能力改变。蛋白质印迹法检测转移相关基因基质金属蛋白酶-2(matrix metallopro teinase-2,MMP-2)和基质金属蛋白酶-9(matrix metalloproteinase-9,MMP-9)表达改变。结果:成功建立了Erbin基因沉默乳腺癌细胞模型。蛋白质印迹法显示,稳定表达Erbin-shRNA细胞的Erbin蛋白表达明显被抑制,Erbin基因沉默后乳腺癌细胞变长、变梭,细胞间隙明显增宽,侵袭能力为56.3±7.6,明显高于空载体转染组的29.3±2.1,t=-5.908,P=0.004;迁移能力为83.6±7.2,较空载体转染组的52.6±8.3显著增强,t=-4.868,P=0.008。Erbin-shRNA转染后转移相关基因MMP-2表达为0.74±0.021,较空载体组的0.25±0.039明显上调,t=-19.23,P<0.001;MMP-9的表达为0.78±0.037,较空载体组的0.32±0.078也显著上调,t=-10.02,P=0.001。结论:Erbin基因的表达沉默对乳腺癌细胞的部分恶性生物学行为的发生有一定促进作用,可促进细胞变长、变梭,促进细胞侵袭迁移能力增强。
OBJECTIVE：To study the effect of RNA interference targeting Erbin gene on the biological behavior of breast carcinoma cells. METHODS： Erbin-ShRNA was transfected into MDA-MB-231 cells to construct an Erbin knockdown cell model. The change of cell morphology was observed under inverted microscope. The change of the expression of Erbin protein was subsequently detected by Western bloting. The change of the effect of cell migration was tested by Tr- answell chambers. The change of the effect of cell invasion was assessed by Matrigel-Transwell. The change of the expression of metastasis-related marker matrix metalloproteinase-2 （MMP-2） and metastasis-related marker matrix metallopro- teinase-9 （MMP-9） were detected by Western blotting. RESULTS.. Erbin-ShRNA resulted in remarkable suppression of Erbin expression in the breast carcinoma cells. Cells were longer and more shuttle, the gap was widened. Meanwhile, the invasion （56.3±7.6 vs 29.3±2.1,t=-5. 908,P=0. 004） and migration （83.6±7.2 vs 52.6±8.3,t=--4. 868,P= 0. 008） potential of MDA-MB-231 cells were enhanced after Erbin gene silencing,and the expression of metastasis-related marker matrix MMP-2 （0. 74 ± 0. 021 vs 0. 25 ± 0. 039, t = - 19. 23, P=0. 001）, metastasis-related marker MMP-9 （0.78±0. 037 vs 0. 32±0. 078, t = -10.02, P = 0. 001） were significantly amplified. CONCLUSION.. Down-regulation of Erbin gene expression can promote malignant biological behavior of breast carcinoma cells, which can promote cells becoming longer and more shuttle,can enhance the invasion and migration potential of MDA-MB-231 cells.
出处
《中华肿瘤防治杂志》
CAS
北大核心
2013年第17期1291-1294,共4页
Chinese Journal of Cancer Prevention and Treatment
基金
国家自然科学基金(30871172
30971372)
