舌鳞状细胞癌组织中FIP200与ATG7的表达水平研究

Study on the expression of FIP200 and ATG7 intissues of tongue squamous cell carcinoma

目的通过免疫组化法检测舌鳞状细胞癌(tongue squamous cell carcinoma, TSCC)患者癌组织及癌旁组织中的200 kDA的家族相互作用蛋白(family interacting protein of 200 kDA,FIP200)、自噬相关基因7(autophagy related gene 7,ATG7)的表达,并分析FIP200、ATG7在不同分组的TSCC中的表达情况,为TSCC的靶向治疗提供前期实验依据。方法选取2012年~2019年就诊的36例TSCC手术患者的石蜡标本用于FIP200免疫组化染色;34例TSCC手术患者的石蜡标本用于ATG7免疫组化染色。以癌组织作为实验组,同一切片的癌旁组织作为对照组,随机选取镜下视野求出面密度值作为FIP200、ATG7的表达情况。收集对应患者的临床资料并分析FIP200、ATG7表达与患者临床病理特征的关系。使用STRING网站构建蛋白质-蛋白质相互作用网络,分别分析与ATG7、FIP200相互作用最为密切的10个蛋白,使用生物信息学分析软件FunRich对2个基因集分别进行生物信息学富集分析。结果 ATG7及FIP200在TSCC组织中的表达高于癌旁组织(P<0.05);ATG7在不同分化等级的癌组织中的表达水平不同,分化等级越高,ATG7的表达水平越低(P<0.05);与ATG7相互作用的10个蛋白分别是GABARAPL2、MAP1LC3B、BECN1、ATG10、GABARAP、ATG12、GABARAPL1、ATG3、MAP1LC3A和ATG5,ATG7作为RIG-I/MDA5信号通路的负调节因子发挥作用(P<0.05);与FIP200相互作用的10个蛋白分别是BECN1、ULK2、ULK1、PIK3C3、ATG101、ATG13、ATG16L1、ATG14、PIK3R4和ATG5,FIP200主要参与细胞中ULK1-ATG13-FIP200复合体以及自噬小体的组成(P<0.001)。结论 ATG7及FIP200在TSCC组织中的表达高于癌旁正常组织;ATG7表达与TSCC的恶性程度有关;ATG7作为RIG-I/MDA5信号通路的负调节因子在生物学过程中发挥作用;FIP200参与ULK1-ATG13-FIP200复合体以及自噬小体的组成,对细胞自噬产生影响。

Objective To detect the expressions of family interacting protein of 200 kDa(FIP200)and autophagy related gene 7(ATG7)in tissues and paracancerous tissues of tongue squamous cell carcinoma(TSCC)patients by immunohistochemistry,and to analyze the expressions of FIP200 and ATG7 in different groups of TSCC,so as to provide a preliminary experimental basis for targeted treatment of TSCC.Methods Paraffin specimens of 36 TSCC patients admitted to hospital from 2012 to 2019 were used for FIP200 immunohistochemical staining,and paraffin specimens of 34 TSCC patients were used for ATG7 immunohistochemical staining.Cancer tissues were used as experimental group,paracancerous tissues in the same sections were used as control group,and the area density value of the field of vision under microscope was randomly selected as the expressions of FIP200 and ATG7.The clinical data of the corresponding patients were collected,and the relationship between the expressions of FIP200 and ATG7 and clinicopathological characteristics was analyzed.Protein-protein interaction network was constructed by STRING website,and 10 proteins that interacted most closely with ATG7 and FIP200 were analyzed,respectively.FunRich(a bioinformatics analysis software)was used to conduct bioinformatics enrichment analysis on two gene sets,respectively.Results The expressions of ATG7 and FIP200 in TSCC tissues were higher than those in paracancerous normal tissues(P<0.05);the expression levels of ATG7 were different in cancer tissues with different grades of differentiation,the higher the grade of differentiation,the lower the expression level of ATG7(P<0.05);10 proteins that interacted with ATG7 were GABARAPL2,MAP1LC3B,BECN1,ATG10,GABARAP,ATG12,GABARAPL1,ATG3,MAP1LC3A and ATG5,and as a negative regulator of RIG-I/MDA5 signaling pathway,ATG7 played a role(P<0.05);10 proteins that interacted with FIP200 were BECN1,ULK2,ULK1,PIK3C3,ATG101,ATG13,ATG16L1,ATG14,PIK3R4 and ATG5,and FIP200 was mainly involved in the composition of ULK1-ATG13-FIP200 complex and autophagosome(P<0.001).Conclusion The expressions of ATG7 and FIP200 in TSCC tissues are higher than those in paracancerous normal tissues,and the expression of ATG7 is related to the malignant degree of TSCC.As a negative regulator of RIG-I/MDA5 signaling pathway,ATG7 plays a role in biological processes.FIP200 is involved in the composition of ULK1-ATG13-FIP200 complex and autophagosome,and affects autophagy.