Interplay between intergrin-linked kinase and ribonuclease inhibitor affects growth and metastasis of bladder cancer through signaling ILK pathways.
Interplay between intergrin-linked kinase and ribonuclease inhibitor affects growth and metastasis of bladder cancer through signaling ILK pathways.
复制标题
整合素连接激酶和核糖核酸酶抑制剂之间的相互作用通过ILK信号通路影响膀胱癌的生长和转移
DOI：
10.1186/s13046-016-0408-x
复制
发表时间：
2016-08-30
期刊：
影响因子：
--
通讯作者：
Chen J
中科院分区：
文献类型：
--
作者：
Zhuang X;Lv M;Zhong Z;Zhang L;Jiang R;Chen J
整合素连接激酶（ILK）是一种多功能衔接蛋白，其参与细胞内的蛋白质信号传导以调节恶性（癌）细胞运动、细胞周期、转移和上皮-间质转化（EMT）。我们的前期实验表明ILK siRNA能抑制膀胱癌细胞的生长并诱导其凋亡，同时还能增加核糖核酸酶抑制因子（Ribonuclease Inhibitor，RI）的表达，RI是一种重要的细胞质蛋白，具有多种功能。我们还报道了RI过表达抑制ILK以及AKT和GSK 3 β的磷酸化。ILK和RI基因均位于染色体11 p15上，且在进化过程中，这两个基因总是位于同一条染色体的相邻位置，这表明ILK和RI可能存在一定的亲缘关系。然而，它们之间潜在的相互作用机制仍不清楚。我们推测RI可能通过与ILK相互作用来调节ILK信号通路。采用免疫共沉淀法、GST pull-down法和激光共聚焦显微镜共定位法检测ILK与RI的内源性和外源性相互作用。此外，我们进一步证实，有两个蛋白质之间的直接结合，通过荧光共振能量转移（FRET）在细胞中。采用Western blot、免疫组化和免疫荧光法检测ILK和RI相互作用对PI 3 K/AKT/mTOR信号通路关键靶蛋白表达的影响。最后，使用裸鼠异种移植模型评估相互作用。本研究通过GST pull-down、免疫共沉淀（Co-IP）和FRET等方法首次发现ILK在体内外均能与RI联合收割机结合。ILK和RI的蛋白水平在体内和体外均呈显著负相关。结果表明，上调ILK表达可促进细胞增殖，改变细胞形态，调节细胞周期。我们还证明，ILK的过表达显着促进EMT和ILK信号通路的靶分子的表达在体外和体内。最后，我们发现ILK过表达显著促进了异种移植瘤的生长、转移和血管生成;而RI在体内和体外的作用与ILK相反。本研究首次直接证明了ILK和RI通过ILK/PI 3 K/AKT信号通路相互作用调节膀胱癌EMT，提示ILK/RI可能是膀胱癌治疗和诊断的重要标志物。
Integrin-linked kinase (ILK) is a multifunctional adaptor protein which is involved with protein signalling within cells to modulate malignant (cancer) cell movement, cell cycle, metastasis and epithelial–mesenchymal transition (EMT). Our previous experiment demonstrated that ILK siRNA inhibited the growth and induced apoptosis of bladder cancer cells as well as increased the expression of Ribonuclease inhibitor (RI), an important cytoplasmic protein with many functions. We also reported that RI overexpression inhibited ILK and phosphorylation of AKT and GSK3β. ILK and RI gene both locate on chromosome 11p15 and the two genes are always at the adjacent position of same chromosome during evolution, which suggest that ILK and RI could have some relationship. However, underlying interacting mechanisms remain unclear between them. Here, we postulate that RI might regulate ILK signaling pathway via interacting with ILK. Co-immunoprecipitation, GST pull-down and co-localization under laser confocal microscope assay were used to determine the interaction between ILK and RI exogenously and endogenously. Furthermore, we further verified that there is a direct binding between the two proteins by fluorescence resonance energy transfer (FRET) in cells. Next, The effects of interplay between ILK and RI on the key target protein expressions of PI3K/AKT/mTOR signaling pathway were determined by western blot, immunohistochemistry and immunofluorescence assay in vivo and in vitro. Finally, the interaction was assessed using nude mice xenograft model. We first found that ILK could combine with RI both in vivo and in vitro by GST pull-down, co-immunoprecipitation (Co-IP) and FRET. The protein levels of ILK and RI revealed a significant inverse correlation in vivo and in vitro. Subsequently, The results showed that up-regulating ILK could increase cell proliferation, change cell morphology and regulate cell cycle. We also demonstrated that the overexpression of ILK remarkably promoted EMT and expressions of target molecules of ILK signaling pathways in vitro and in vivo. Finally, we found that ILK overexpression significantly enhanced growth, metastasis and angiogenesis of xenograft tumor; Whereas, RI has a contrary role compared to ILK in vivo and in vitro. Our findings, for the first time, directly proved that the interplay between ILK and RI regulated EMT via ILK/PI3K/AKT signaling pathways for bladder cancer, which highlights the possibilities that ILK/RI could be valuable markers together for the therapy and diagnosis of human carcinoma of urinary bladder.
登录
查看更多内容
影响因子：
16.6
作者：
Conde-Perez A;Gros G;Longvert C;Pedersen M;Petit V;Aktary Z;Viros A;Gesbert F;Delmas V;Rambow F;Bastian BC;Campbell AD;Colombo S;Puig I;Bellacosa A;Sansom O;Marais R;Van Kempen LC;Larue L
通讯作者：
Larue L
靶向整合素连接激酶的小干扰RNA抑制人膀胱癌细胞的生长并诱导细胞凋亡
DOI：
10.1016/j.biocel.2011.05.003
发表时间：
2011-09-01
影响因子：
4
作者：
Gao, Juan;Zhu, Jun;Chen, Jun-Xia
通讯作者：
Chen, Jun-Xia
影响因子：
3.4
作者：
Datta, Dhrubajyoti;Dasgupta, Swagata;Pathak, Tanmaya
通讯作者：
Pathak, Tanmaya
影响因子：
2.7
作者：
Li, Wen-Lei;Zhang, Yang;Tian, Xing-Song
通讯作者：
Tian, Xing-Song
影响因子：
3.5
作者：
Monti, Daria Maria;Gesualdi, Nicola Montesano;D'Alessio, Giuseppe
通讯作者：
D'Alessio, Giuseppe