摘要
目的探讨ARK5在乳腺癌侵袭和转移中的意义。方法采用免疫组化SP法检测58例乳腺癌组织和15例癌旁乳腺组织中MMP-2、MMP-9及ARK5表达。应用小RNA干扰技术,将合成的小RNA干扰质粒转染MDA-MB-435细胞株,采用Western blot法检测瞬时转染后ARK5蛋白表达情况。通过体外侵袭实验检测细胞转染后侵袭能力的变化。ARK5下降后,再次进行Western blot法检测MMP-2、MMP-9蛋白表达。结果乳腺癌组织中MMP-2、MMP-9和ARK5免疫组化阳性结果之间存在正相关性。转染后的细胞株命名:转染ARK5质粒的MDA-MB-435细胞称之为SiARK5/MDA-MB-435,转染对照质粒的MDA-MB-435细胞称之为Scr/MDA-MB-435。转染72 h后,与Scr/MDA-MB-435细胞相比,SiARK5/MDA-MB-435细胞的ARK5蛋白表达水平降低。ARK5表达降低的乳腺癌细胞侵袭并穿透Matrivgel膜基质的细胞数量比对照组少(P<0.01),且MMP-2、MMP-9蛋白表达量降低。结论应用小RNA干扰技术降低ARK5蛋白的表达使MDA-MB-435细胞侵袭转移能力降低,同时MMP-2、MMP-9蛋白表达降低,提示ARK5通过MMP-2、MMP-9在乳腺癌侵袭转移中发挥重要作用。
Purpose To study the effect and significance of ARK5 in the invasiveness and metastasis of breast cancer.Methods Immunohistochemistry was used to detect the expression of MMP-2,MMP-9 and ARK5 in 58 cases of breast cancer and 15 cases of adjacent nontumor tissue.SiRNA plasmid was used to transfect MDA-MB-435 cells.Western blot was used to analyze protein expression of ARK5.In vitro Matrigel invasion assay was used to detect the variation of invasiveness in the cells being transfected.Western blot was used to analyze protein expression of MMP-2 and MMP-9.Results The expression of ARK5 in breast cancer was significantly related with MMP-2 and MMP-9.MDA-MB-435 cells that has been transfected by ARK5 plasmid was named SiARK5/MDA-MB-435 and MDA-MB-435 cells that has been transfected by control plasmid was named Scr/MDA-MB-435.72 hours after transfection,the protein expression of ARK5 in SiARK5/MDA-MB-435 was decreased.After downregulation of ARK5 the quantity of breast cancer cells which invaded and penetrated Matrivgel was decreased（P0.01）.Conclusion Silencing of ARK5 impairs breast cancer cell invasion,suggesting that ARK5 can play a key role in the invasiveness and metastasis of breast cancer by adjusting the expression of MMP-2 and MMP-9.
出处
《临床与实验病理学杂志》
CAS
CSCD
北大核心
2012年第4期382-385,共4页
Chinese Journal of Clinical and Experimental Pathology
基金
国家自然科学基金(81072068)
山东省中青年科学家科研奖励基金(博士基金)(2010BSB14050)
