Chemerin过表达通过TGF-β/Smad信号通路及中性粒细胞类型转化促进舌鳞癌演进的研究

Previous researches have showed that the infiltrated neutrophils in tumors maybe a strong promoter for tumor invasion and metastasis, and its type can be converted under the effect of some molecules in tumor microenvironment, such as IFN-γ或TGF-βetc. While the regulatory mechanisms of its infiltration and transformation was still unclear. Our previous research have found that there were abundant neutrophils infiltration in tongue squamous cell carcinoma (TSCC) and its infiltration was associated with poor clinical outcomes. We also found that Chemerin, a newly discovered adipokine which has chemotaxis to some inflammatory cells, was upregulated in TSCC tissues, and was positively related to poor prognosis of patients and TGF-β1 expression. Based on these, we speculate that the overexpressed Chemerin in TSCC may have regulatory function on TSCC progression. Chemerin may attract neutrophils to tumor sites, which express its receptors, and promote transformation of neutrophils from antitumoral type to protumoral type through the TGF-β/Smad signaling pathway, and so as to further accelerate tumor progression. In this study, technologies of gene interference, coculture, nude-mouse transplanted tumor model, PCR, Western etc will be used to explore the possible roles of Chemerin on TSCC cells and on the infiltration, function and transformation of neutrophils, and their molecular mechanisms. This research will provide new target molecule and theoretical basis for comprehensive antitumoral strategy of oral carcinoma.

研究表明肿瘤组织内浸润的中性粒细胞可能是肿瘤侵袭、转移的重要促进因素，并且在肿瘤微环境中IFN-γ或TGF-β1等因子作用下可发生类型转化，但其浸润及类型转化的调控机制尚不明确。我们前期研究发现，舌癌组织中存在着中性粒细胞浸润，并且与不良的临床结局有关。我们也发现可发挥趋化作用的新脂肪因子Chemerin在舌癌组织中表达上调，并与较差预后及TGF-β1的表达呈正相关。基于以上，我们推测舌癌高表达的Chemerin可能在舌癌演进过程中发挥调控作用，能趋化表达其受体的中性粒细胞，通过调控TGF-β/Smad信号通路促进中性粒细胞向促肿瘤类型转化，从而进一步促进舌癌的演进和发展。本项目拟采用基因干扰、共培养、裸鼠模型、PCR、Western等技术研究Chemerin过表达对舌癌细胞自身及对中性粒细胞浸润、功能和类型转化的影响和分子机制。该研究将为口腔癌的全面抗肿瘤策略提供新的靶分子及理论依据。

研究表明肿瘤组织内浸润的中性粒细胞可能是肿瘤侵袭、转移的重要促进因素，并且在肿瘤微环境作用下可能发生类型转化，由抗肿瘤类型向促肿瘤类型转换，然而其浸润及类型转化的调控机制尚不明确。.研究内容：1.探究了舌鳞癌组织内chemerin过表达与中性粒细胞浸润的相关性，以及chemerin/ChemR23通路对中性粒细胞的趋化作用；2.探究了舌鳞癌组织内chemerin表达对肿瘤细胞自身生物学行为的作用； 3.研究了中性粒细胞与舌癌细胞共培养对中性粒细胞类型转化的影响，以及TGF-β信号通路在中性粒细胞类型转化中的作用。4.探究chemerin/ChemR23通路对中性粒细胞功能的影响，以及中性粒细胞对舌癌细胞的反哺作用。.研究结果：1.免疫组化双染结果显示：舌癌组织内chemerin表达与中性粒细胞密度呈正相关。体外Transwell实验提示，表达chemerin的舌癌细胞及人重组chemerin均可通过chemerin/ChemR23通路对诱导中性粒细胞产生趋化作用；2.体外功能学实验显示，舌癌细胞内chemerin过表达可以促进肿瘤细胞增殖、侵袭和迁移能力；3.共培养实验表明：诱导中性粒细胞与舌癌细胞共培养后，可由抗肿瘤类型向促肿瘤类型转换，并且该转换作用主要是通过TGF-β信号通路实现；4.在人重组chemerin作用下，诱导中性粒细胞与舌癌细胞共培养后，可以促进肿瘤细胞的克隆形成、增殖和迁移及侵袭；5.应用人重组chemerin后，诱导中性粒细胞内IL-5和IL-17等的表达水平明显升高，提示chemerin可能会通过中性粒细胞促进舌癌细胞的EMT。.综上，该课题将有助于阐明舌癌组织内chemerin表达对中性粒细胞浸润及其功能影响的分子机制，丰富中性粒细胞在肿瘤中的作用。靶向chemerin及中性粒细胞可为口腔癌的治疗及预后判定提供新的思路。