STOML2 interacts with PHB through activating MAPK signaling pathway to promote colorectal Cancer proliferation.
STOML2 interacts with PHB through activating MAPK signaling pathway to promote colorectal Cancer proliferation.
复制标题
STOML2通过激活MAPK信号通路与PHB相互作用，促进结直肠癌增殖。
DOI：
10.1186/s13046-021-02116-0
复制
发表时间：
2021-11-15
期刊：
影响因子：
--
通讯作者：
Geng Y
中科院分区：
文献类型：
--
作者：
Ma W;Chen Y;Xiong W;Li W;Xu Z;Wang Y;Wei Z;Mou T;Wu Z;Cheng M;Zou Y;Zhu Y;Zhou W;Liu F;Geng Y
STOML 2在多种肿瘤中的高表达已有报道，但很少有详细的功能和调控机制。本研究旨在通过探讨STOML 2在结直肠癌中的作用，鉴定其相互作用蛋白，揭示其调控机制，为临床治疗提供依据。采用病理学方法检测南方医院生物库正常结肠和结直肠组织中STOML 2的表达水平。通过功能获得性和功能丧失性实验验证了STOML 2诱导的CRC恶性增殖，并应用了新技术，如类器官培养、原位模型和内窥镜监测。利用酵母双杂交技术筛选与STOML 2相互作用的蛋白，并通过生物信息学分析预测候选蛋白的生物学功能和信号通路。通过免疫共沉淀法验证与STOML 2功能最相似的靶蛋白，并进行免疫荧光共定位STOML 2和PHB。免疫印迹法检测STOML 2调控的通路，并进行RAF抑制剂索拉非尼的后续实验治疗。STOML 2在结直肠癌中显著过表达，其升高与不良预后相关。STOML 2的敲低抑制了结直肠癌的增殖，从而减弱了皮下和原位肿瘤的生长，而过表达的STOML 2促进了细胞系和类器官的增殖。利用酵母双杂交技术筛选出13个相互作用蛋白。根据生物信息学，DTYMK和PHB在生物过程和信号通路方面与STOML 2最相似;然而，免疫共沉淀证实了STOML 2和PHB之间的相互作用，而不是DTYMK，尽管它在之前的分析中排名最高。在细胞系和组织水平上证实了STOML 2和PHB之间的共定位。此外，敲低STOML 2下调MAPK信号通路上RAF 1、MEK 1/2和ERK 1/2的磷酸化，表明STOML 2和PHB在结直肠癌增殖中激活共同通路。本研究表明，在大肠癌中，STOML 2表达升高，并通过激活MAPK信号通路与PHB相互作用，促进体外和体内增殖。此外，筛选技术与生物信息学的结合，对研究目的蛋白的调控机制具有重要的方法学意义。在线版本包含补充材料，可通过10.1186/s13046-021-02116-0获得。
Highly expressed STOML2 has been reported in a variety of cancers, yet few have detailed its function and regulatory mechanism. This research aims to reveal regulatory mechanism of STOML2 and to provide evidence for clinical therapeutics, via exploration of its role in colorectal cancer, and identification of its interacting protein. Expression level of STOML2 in normal colon and CRC tissue from biobank in Nanfang Hospital was detected by pathologic methods. The malignant proliferation of CRC induced by STOML2 was validated via gain-of-function and loss-of-function experiments, with novel techniques applied, such as organoid culture, orthotopic model and endoscopy monitoring. Yeast two-hybrid assay screened interacting proteins of STOML2, followed by bioinformatics analysis to predict biological function and signaling pathway of candidate proteins. Target protein with most functional similarity to STOML2 was validated with co-immunoprecipitation, and immunofluorescence were conducted to co-localize STOML2 and PHB. Pathway regulated by STOML2 was detected with immunoblotting, and subsequent experimental therapy was conducted with RAF inhibitor Sorafenib. STOML2 was significantly overexpressed in colorectal cancer and its elevation was associated with unfavorable prognosis. Knockdown of STOML2 suppressed proliferation of colorectal cancer, thus attenuated subcutaneous and orthotopic tumor growth, while overexpressed STOML2 promoted proliferation in cell lines and organoids. A list of 13 interacting proteins was screened out by yeast two-hybrid assay. DTYMK and PHB were identified to be most similar to STOML2 according to bioinformatics in terms of biological process and signaling pathways; however, co-immunoprecipitation confirmed interaction between STOML2 and PHB, rather than DTYMK, despite its highest rank in previous analysis. Co-localization between STOML2 and PHB was confirmed in cell lines and tissue level. Furthermore, knockdown of STOML2 downregulated phosphorylation of RAF1, MEK1/2, and ERK1/2 on the MAPK signaling pathway, indicating common pathway activated by STOML2 and PHB in colorectal cancer proliferation. This study demonstrated that in colorectal cancer, STOML2 expression is elevated and interacts with PHB through activating MAPK signaling pathway, to promote proliferation both in vitro and in vivo. In addition, combination of screening assay and bioinformatics marks great significance in methodology to explore regulatory mechanism of protein of interest. The online version contains supplementary material available at 10.1186/s13046-021-02116-0.
登录
查看更多内容
DOI：
10.4049/jimmunol.181.3.1927
发表时间：
2008-08-01
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
Kirchhof MG;Chau LA;Lemke CD;Vardhana S;Darlington PJ;Márquez ME;Taylor R;Rizkalla K;Blanca I;Dustin ML;Madrenas J
通讯作者：
Madrenas J
影响因子：
5.2
作者：
Chen, Min-Shan;Lo, Yuan-Hung;Shroyer, Noah F.
通讯作者：
Shroyer, Noah F.
影响因子：
3.7
作者：
Christie DA;Kirchhof MG;Vardhana S;Dustin ML;Madrenas J
通讯作者：
Madrenas J
DOI：
10.1016/s0140-6736(17)33326-3
发表时间：
2018-03-17
期刊：
Lancet (London, England)
影响因子：
--
作者：
Allemani C;Matsuda T;Di Carlo V;Harewood R;Matz M;Nikšić M;Bonaventure A;Valkov M;Johnson CJ;Estève J;Ogunbiyi OJ;Azevedo E Silva G;Chen WQ;Eser S;Engholm G;Stiller CA;Monnereau A;Woods RR;Visser O;Lim GH;Aitken J;Weir HK;Coleman MP;CONCORD Working Group
通讯作者：
CONCORD Working Group
DOI：
10.26355/eurrev_202002_20340
发表时间：
2020-02-01
影响因子：
3.3
作者：
Guo, X-Y;Guo, H-F;Guo, H-M
通讯作者：
Guo, H-M