摘要
目的:探讨siRNA干扰降低腺苷酸活化蛋白激酶5(AMPK-related protein kinase 5,ARK5)表达对U251细胞侵袭潜能的影响及其机制。方法:采用蛋白质印迹法检测U251细胞中ARK5表达。应用siRNA干扰技术转染U251细胞株,并采用蛋白质印迹法检测瞬时转染后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,蛋白质印迹法检测间质细胞来源的YKL-40、N-cadherin、Vimentin蛋白和调节因子Snail、Slug、Twist1和Zeb1的表达。结果:转染ARK5质粒的U251细胞ARK5表达下降。ARK5表达降低的U251细胞侵袭并穿透Matrigel膜基质的细胞数量为22,U251对照组为41,SCR/U251对照组为42,两两比较,ARK5表达降低的实验组透膜数比两对照组少(P<0.001),同时蛋白质印迹法结果显示,YKL-40、N-cadherin和Vimentin蛋白表达量降低,与间质转化相关的转录因子仅Snail的表达降低,而转录因子Slug、Twist1和Zeb1表达无明显改变。结论:应用siRNA干扰技术降低ARK5蛋白的表达使U251细胞侵袭转移能力降低,同时YKL-40、N-cadherin和Vimentin蛋白和转录因子Snail的表达量降低,提示ARK5蛋白表达降低可通过调节间质转化影响U251细胞的侵袭。
OBJECTIVE： To investigate the influence of invasion potentiality and its mechanism for U251 cell by siR- NA interference reducing ARK5. METHODS： Western blot was used to detect the expression of ARK5 in U251 ceils. SiR- NA plasmid was used to transfect U251 cells, then 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 YKL-40, N-cadherin, Vimentin and expression of transcription factor Snail, Slug, Twistl ,Zebl respectively. RESULTS： The protein expression of ARK5 in U251 cells that had been transfected by ARK5 was decreased. After downregulation of ARKS,the quantity of U251 cells which invaded and penetrated matriget was 22. The blank control group was 41, the control group of SCR/U251 was 42, compared with the two control groups, the quan- tity of U251 cells and the downregulation of ARK5 were decreased （P^0. 001）. The protein expression of YKL-40, N-cadherin,Vimentin and only expression of transcription factor Snail were decreased significantly. CONCLUSIONS： Silen- cing of ARK5 impairs glioma cell invasion and the expression of ARK5 in glioma was signifieantly related with the expres- sion of YKL-40, N-cadherin,：Vimentin and Snail, suggesting that ARK5 can play a key role in the invasiveness of U251 by adjusting MT.
出处
《中华肿瘤防治杂志》
CAS
北大核心
2014年第3期161-164,共4页
Chinese Journal of Cancer Prevention and Treatment
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(2010BSB14050)
