BMK1表达对胶质瘤细胞U87侵袭能力影响机制探讨  

Effect and mechanism of BMK1 in the invasion of glioma cells U87

作　　者：齐岳亮 陈为一[1] 李小龙[1] 陈萍萍[1] 李洪利[2] 刘晓丽[3] 张宝刚[1] 

机构地区：[1]潍坊医学院病理学教研室,山东潍坊261053 [2]潍坊医学院医学研究实验中心,山东潍坊261053 [3]潍坊医学院护理学院,山东潍坊261053

出　　处：《中华肿瘤防治杂志》2014年第21期1667-1670,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(博士)基金(2010BSB14050)

摘　　要：目的探讨大丝裂原活化蛋白激酶1(big mitogen-activated protein kinase 1,BMK1)表达对U87细胞侵袭能力的影响及作用机制。方法应用siRNA干扰技术转染U87细胞株,并采用蛋白质印迹法检测U87细胞中BMK1表达及转染后BMK1表达情况。体外癌细胞侵袭实验检测U87细胞转染后侵袭能力变化。BMK1下降后,蛋白质印迹法检测间叶细胞特异性标记蛋白N-cadherin、T-cadherin、Vimentin和转录因子Snail、Slug、Twist1、Zeb1的表达。结果蛋白质印迹法检测结果显示,U87细胞中BMK1相对表达量为1.0±0.21,SCR/U87细胞中BMK1相对表达量为1.1±0.18,SiBMK1/U细胞中BMK1相对表达量为0.45±0.12,差异有统计学意义,F=12.129,P=0.008。体外癌细胞侵袭实验显示,SiBMK1/U87实验组穿透Matrigel膜基质的细胞数为106±18,SCR/U87对照组为61±15,U87对照组为62±14,差异有统计学意义,F=7.977,P=0.020;U87对照组与SCR/U87对照组差异无统计学意义,P=0.941;而SiBMK1/U87实验组穿透基膜的细胞数明显多于U87对照组(P=0.014)和SCR/U87对照组(P=0.013),差异有统计学意义。蛋白质印迹法结果显示,SCR/U87和SiBMK1/U87细胞中N-cadherin相对表达量分别为1.0±0.13和3.8±0.31,t=-14.427,P<0.001;Vimentin相对表达量分别为1.5±0.16和3.9±0.22,t=-15.281,P<0.001;T-cadherin相对表达量分别为4.5±0.12和1.3±0.14,t=30.059,P<0.001。SCR/U87和SiBMK1/U87细胞中Twist1蛋白相对表达量分别为0.75±0.14和2.3±0.22,t=-10.295,P=0.001,差异有统计学意义。表明BMK1表达影响Twist1的表达。结论 BMK1与U87细胞侵袭性显著相关,其侵袭性的调控是通过间叶转化(mesenchymal transition,MT)实现的。OBJECTIVE To explored the role and mechanism of BMK1 on the invasion of U87 cells. METHODS siRNA plasmid was used to transfect U87 ceils. Western blot was used to analyze the expression of BMK1 in transfected U87 cells and normal U87 cells. In vitro Matrigel invasion assay was used to detect the variation of invasiveness in trans- fected U87 cells. Western blot was used to analyze the expression of mesenchymal cell-specific markers which include N-cadherin, T-cadherin and Vimentin and transcription factors which include Snail,Slug,Twist1 and Zebl in the BMK1 re- ducing U87 cells. RESULTS The relative expression of BMK1 in U87, SCR/U87 and SiBMK1/U87 cells was respec- tively 1.0±0.21, 1.1±0.18 and 0.45＋0.12. The differences were statistically significant, F=12. 129, P=0. 008. In vitro Matrigel invasion assay showed that the quantity of U87 cells which invaded and penetrated Matrigel was 106 4-18 in SiBMK1/U87cells, 61±15 in SCR/U87 cells and 62! 14 in the normal U87, and the differences were statistically signifi- cant, F= 7. 977, P=O. 020. There was no differences between normal U87 cells and SCR/U87cells, P=0. 941; but the quantity of invaded U87 cells was obvious more in SiBMK1/U87 cells than in U87 cells （P=0. 014） and in SCR/U87 cells （P=0. 013）. Western blot was used to analyze the relative expression of mesenehymal cell specific markers （N-cadherin, T-cadherin and Vimentin） and MT transcription factors（Snail, Slug, Twist1 and Zebl） in SCR/U87 and SiBMK1/U87 cells. N-cadherin was 1.0 ± 0.13 and 3.8 ± 0.31, t =- 14. 427, P〈0. 001; Vimentin was 1.5 ± 0.16 and 3.9±0.22, t = -15. 281, P〈0. 001; T-cadherin was 4.5±0.12 and 1.3＋0.14, t=30. 059, P〈0. 001; Twist1 was 0.75±0.14 and 2.3 ± 0.22, t=- 10. 295, P=0. 001, the differences were statistically significant. CONCLUSION BMK1 is strongly as- sociated with U87 cells invasion, the control of invasion is by mesenchymal transition（MT） implementation.

关 键 词：U87细胞 SIRNA干扰 大丝裂原活化蛋白激酶1 间叶转化 侵袭 

分 类 号：R739.41[医药卫生—肿瘤]