TTK基因在眼睑基底细胞癌中的表达及其对恶性肿瘤细胞生物学行为的影响  

The expression of the threonine and tyrosine kinase gene in eyelid basal cell carcinoma and its impact on the biological behavior of malignant tumor cells

作　　者：李涛 漆星 张丹 张宇茹 张婷婷 郑玲玲 戴传强 唐娟 LI Tao;QI Xing;ZHANG Dan;ZHANG Yuru;ZHANG Tingting;ZHENG Lingling;DAI Chuanqiang;TANG Juan(Department of Ophthalmology,Ziyang Central Hospital,Ziyang 641300,Sichuan Province,China;Key Laboratory of Ophthalmology,Ziyang,Ziyang 641300,Sichuan Province,China;Department of Experimental Medicine,Ziyang Central Hospital,Ziyang 641300,Sichuan Province,China;Department of Medical Education,Ziyang Central Hospital,Ziyang 641300,Sichuan Province,China;Department of Endocrinology,Ziyang Central Hospital,Ziyang 641300,Sichuan Province,China)

机构地区：[1]资阳市中心医院眼科,四川省资阳市641300 [2]资阳市眼科重点实验室,四川省资阳市641300 [3]资阳市中心医院实验医学科,四川省资阳市641300 [4]资阳市中心医院医教部,四川省资阳市641300 [5]资阳市中心医院内分泌科,四川省资阳市641300

出　　处：《眼科新进展》2025年第4期280-285,共6页Recent Advances in Ophthalmology

基　　金：2024年四川省卫生健康委员会科技项目(编号:24WSXT096,24WSXT098);四川省老年医学学会课题计划项目(编号:24SCLN0115);河北省医学科学研究课题计划(编号:20240294);四川省资阳市医学科学课题计划项目(编号:KY2023001,KY2023023);四川省资阳市科学技术局计划项目(编号:zykjjsc20-yyjc-2023-04)。

摘　　要：目的探索苏氨酸和酪氨酸激酶(TTK)基因与眼睑基底细胞癌(BCC)的相关性。方法本研究基于GEO数据库,利用生物信息学方法筛选出与BCC肿瘤发生发展相关的核心基因TTK。收集资阳市中心医院手术切除的眼睑BCC组织标本(随着BCC恶性程度加重,将BCC细胞分为BCC Grade I组、BCC Grade II组及BCC Grade III组)与眼睑良性肿瘤组织标本(设为Control组)进行后续实验比较。采用细胞免疫荧光法(CIA)检测TTK基因在眼睑良性肿瘤细胞和BCC细胞中的表达;慢病毒转染BCC细胞敲低TTK后[分别转染LV-TTK-shRNA(设为TTK-shRNA组)和阴性对照序列LV-BCC-shRNA(设为BCC阴性对照组)],采用CIA检测各组细胞中凋亡信号通路中关键蛋白Bcl-2和Bax蛋白表达情况。结果生物信息学方法筛选出与BCC肿瘤发生和发展相关的核心基因为TTK。CIA检测结果显示,Control组、BCC Grade I组、BCC Grade II组及BCC Grade III组肿瘤细胞质中荧光强度分别为1.03±0.07、1.28±0.11、1.58±0.13及1.92±0.17,荧光强度逐渐增强,各组间细胞荧光强度比较,差异均有统计学意义(均为P<0.05)。Control组、BCC阴性对照组及TTK-shRNA组细胞荧光强度分别为1.02±0.05、1.74±0.12及1.31±0.09。与Control组比较,BCC阴性对照组细胞荧光强度增强,TTK-shRNA组细胞荧光强度降低,三组间互相比较,差异均有统计学意义(均为P<0.05)。Control组、BCC阴性对照组及TTK-shRNA组细胞中,抗凋亡蛋白BcL-2荧光强度分别为1.04±0.12、2.12±0.23及1.43±0.15;促凋亡蛋白Bax荧光强度分别为1.02±0.08、0.64±0.11及1.47±0.16。TTK敲低后,BCC细胞中BcL-2表达水平降低,Bax表达水平升高,三组间BcL-2及Bax荧光强度组间互相比较,差异均有统计学意义(均为P<0.05)。结论TTK基因参与了眼睑BCC细胞的增殖调控,并且这种作用与PI3K-AKT-Bcl-2/Bax信号通路密切相关。Objective To investigate the relationship between the threonine and tyrosine kinase(TTK)gene and eyelid basal cell carcinoma(BCC).Methods Bioinformatics methods were used to screen the core gene(namely,TTK)associated with the occurrence and development of BCC from the Gene Expression Omnibus(GEO)database.Surgically removed eyelid BCC tissue specimens(BCC cells were divided into BBC GradeⅠ,Ⅱ,andⅢgroups by tumor grade)and benign tumor tissue specimens(Control group)were collected from Ziyang Central Hospital for subsequent experiments.Cellular immunofluorescence assay(CIA)was used to detect the expression of the TTK gene in benign and malignant eyelid tumor cells.After knocking down TTK in BCC cells through transfection with lentiviruses(the cells transfected with LV-TTK-shRNA were taken as the TTK-shRNA group,and those transfected with LV-BBC-shRNA were taken as the BBC negative control group),CIA was used to detect the expression of key proteins Bcl-2 and Bax in the apoptotic signaling pathway of each group of cells.Results The bioinformatics analysis showed that the TTK gene was the core gene associated with the occurrence and development of eyelid BCC.CIA detection results revealed that the fluorescence signal intensities in the tumor cytoplasm of Control,BCC GradeⅠ,Ⅱ,andⅢgroups were 1.03±0.07,1.28±0.11,1.58±0.13 and 1.92±0.17,respectively.The fluorescence signal intensity gradually increased,and the difference in fluorescence signal intensity among the four groups was statistically significant(all P<0.05).Compared with that in the Control group(1.02±0.05),the cell fluorescence intensity was increased in the BCC negative control group(1.74±0.12)and decreased in the TTK-shRNA group(1.31±0.09)(P<0.05).The difference in cell fluorescence intensity was significant among the Control,BCC negative control and TTK-shRNA groups(all P<0.05).The fluorescence intensity of the anti-apoptotic protein Bcl-2 was 1.04±0.12 in the Control group,2.12±0.23 in the BCC negative control group,and 1.43±0.15 in th

关 键 词：眼睑基底细胞癌 生物医学信息 细胞免疫荧光法 苏氨酸和酪氨酸激酶 Bcl-2 BAX 

分 类 号：R777.1[医药卫生—眼科]