摘要
目的研究核糖核酸酶抑制因子(RI)与整合素连接激酶(ILK)相互作用对膀胱癌上皮-间质转化(EMT)与转移的影响。方法构建过表达RI或ILK的膀胱癌EJ细胞系,免疫荧光检测RI与ILK在EJ细胞中的表达及共定位,倒置相差显微镜下观察细胞形态,Western blot检测细胞中RI、ILK及EMT标志物的表达,复制膀胱癌裸鼠移植瘤模型,免疫组织化学分析瘤组织中RI、ILK及EMT标志物的表达,HE染色观察裸鼠肺转移。结果 EJ细胞中RI与ILK存在共定位的现象,RI与ILK存在相互作用。Western blot结果显示,EJ-RI组E-钙黏蛋白(E-cadherin)表达高于EJ组(P<0.05),EJ-ILK组基质金属蛋白酶2(MMP2)、基质金属蛋白酶9(MMP9)、N-钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)、扭转蛋白(Twist)、核转录因子(Snail)与重组人S100钙结合蛋白A4(S100A4)表达高于EJ组(P<0.05)。过表达RI抑制膀胱癌体内外EMT及移植瘤自发性肺转移,而过表达ILK促进EMT发生及膀胱癌转移。结论 RI与ILK相互作用调节膀胱癌EMT与转移。
Objective To investigate the regulative effect of crosstalk between ribonuclease inhibitor（RI） and intergrin-linked kinase（ILK） on epithelial-mesenchymal transition（EMT） and metastasis of bladder cancer. Methods Bladder cancer cell lines with overexpression of RI or ILK were constructed. Western blot was performed to analyze the expressions of RI, ILK and EMT markers. Immunofluorescence assay was performed to identify colocalization of RI and ILK. Cell morphology was observed under phase contrast microscope. The model of bladder cancer xenografts in nude mice was established. HE staining of lung tissues was conducted to evaluate pulmonary metastasis of bladder cancer. Results The co-localization and interaction of RI and ILK were observed and verified in EJ cells. Western blot showed that the expression of E-cadherin was significantly increased in EJ-RI cells compared with that in EJ cells. Expression levels of MMP-2, MMP-9, N-cadherin, Vimentin, Twist, Snail and S100 A4 in EJILK cells were dramatically upregulated when compared with EJ cells. Overexpression of RI suppressed EMT and metastasis both in vitro and in vivo. Overexpression of ILK promoted EMT and metastasis both in vitro and in vivo. Conclusion Crosstalk between RI and ILK regulates EMT and metastasis of bladder cancer.
作者
舒静
姜源
庄翔
陈俊霞
Jing Shu;Yuan Jiang;Xiang Zhuang;Jun-Xia Chen(Department of Laboratory Medicine,Afliated Hospital of Southwest Medical University,Luzhou,Sichuan 646000,China;Department of Cell Biology and Genetics,Chongqing Medical University,Chongqing 400016,China)
出处
《中国现代医学杂志》
CAS
2018年第18期1-6,共6页
China Journal of Modern Medicine
基金
国家自然科学基金(No:81672536
81372203)
