肝细胞癌变过程中Wnt3a过表达及其早期监测价值  

Alteration of Wnt3a overexpression and its early monitoring value during hepatocellular carcinogenesis

作　　者：姚敏[1,2] 王建军 王玲玲[2] 秦艳艳 赛文莉[1] 王理[2] 沈水杰 姚登福[1] Yao Min;Wang Jianjun;Wang Lingling;Qin Yanyan;Sai Wenli;Wang Li;Shen Shuijie;Yao Dengfu(Research Center of Clinical Medicine,Affiliated Hospital of Nantong University,Nantong 226001,China;Department of Immunology,Medical School of Nantong University,Nantong 226001,China;Jiangsu Nantong Higher Vocational and Technical School of Health,Nantong 226016;Department of Oncology,Nantong Hospital of Traditional Chinese Medicine,Nantong 226001,China)

机构地区：[1]南通大学附属医院临床医学研究中心,南通226001 [2]南通大学医学免疫学系,南通226001 [3]江苏省南通卫生高等职业技术学校,南通226016 [4]南通市中医院肿瘤内科,南通226001

出　　处：《中华肝脏病杂志》2022年第9期947-953,共7页Chinese Journal of Hepatology

基　　金：国家自然科学基金(81873915,31872738,81673241);南通市重点专项(MS12020021);南通大学医学科技项目(TDYX2021010)。

摘　　要：目的以肝癌发生模型探讨癌胚型Wnt3a动态表达规律及其早期监测价值。方法Sprague-Dawley(SD)鼠48只,以含2-乙酰氨基芴(2-FAA,0.05%)颗粒饲料喂养,诱发肝癌发生,对照鼠以颗粒饲料喂养,每隔2周留取肝组织和血液;肝组织经HE染色病理学检查并分组。以全基因组芯片分析基因和Wnt3a mRNA表达情况,以免疫组织化学分析Wnt3a在肝组织的表达分布情况,以酶联免疫吸附法定量肝组织和血清Wnt3a浓度。采用χ2检验、Mann-Whitney检验和方差分析等统计学方法分析组间差异。结果依据病理学检查,将鼠肝分为对照、肝细胞变性、癌前病变和肝癌形成4组。全基因表达谱分析并与对照组比较,在信号对数比率(SLR)>8 log2cy5/cy3时,癌前组和癌变组分别有268个、312个基因上调,57个、201个基因下调,这些显著改变的基因主要涉及细胞增殖、信号转导、肿瘤转移、细胞凋亡等。诱癌各阶段Wnt3a在mRNA水平表达增加,变性组(1.15±0.24,q=8.227)、癌前组(1.85±0.18,q=12.361)和肝癌组(2.59±0.55,q=18.082)均显著高于对照组(0.25±0.11,F=121.103,P<0.001),变性组、癌前组和肝癌组分别上调4.6倍、7.4倍和10.4倍。免疫组织化学显示Wnt3a表达与对照组比较,变性组阳性率为66.7%(12/18,χ2=10.701,P=0.001)。癌前组和肝癌组均全数阳性(9/9,χ2=17.115,P<0.001)。癌变过程中肝及血中Wnt3a呈进行表达增加,组间差异有统计学意义(F肝=176.711,P<0.001);癌变肝组织表达的Wnt3a分泌入血,血与肝Wnt3a水平呈直线相关(r=0.732,P<0.001)。结论Wnt3a过表达与肝细胞癌变密切相关,有望成为监测肝细胞癌变新标志物。Objective To investigate the dynamic expression pattern of carcinoembryonic Wnt3a and its early monitoring value using a hepatocellular carcinoma model.Methods Forty-eight Sprague Dawley(SD)rats were fed with pellet feed containing 2-acetylaminofluorene(2-AAF,0.05%)to induce hepatocarcinogenesis,and control rats were fed a pellet diet.Liver tissue and blood samples were collected every two weeks.Liver tissues were pathologically examined using HE staining and grouped.The gene and Wnt3a mRNA expression were analyzed by genome-wide microarray.The expression and distribution of Wnt3a in liver tissue were analyzed by immunohistochemistry.Wnt3a concentration in liver tissue and serum was quantified by enzyme-linked immunosorbent assay.Statistical methods such asχ2 test,Mann-Whitney test and analysis of variance were used to analyze the differences between groups.Results According to the pathological examination results,the rat livers were divided into four groups:control,hepatocyte degeneration,precancerous lesions and hepatocellular carcinoma.Genome-wide expression profiling analysis and comparison with the control group revealed that 268 and 312 genes were up-regulated and 57 and 201 genes were down-regulated in the precancerous and cancerous group when signal logarithm ratio(SLR)was>8 log2cy5/cy3,and these significantly altered genes mainly involved in cell proliferation,signal transduction,tumor metastasis,and apoptosis.The expression of Wnt3a at mRNA level was significantly increased in all stages of cancer induction,including degeneration group(1.15±0.24,q=8.227),precancerous group(1.85±0.18,q=12.361)and cancerous group(2.59±0.55,q=18.082).Compared with the control group(0.25±0.11,F=121.103,P<0.001),the degeneration group,the precancerous group and the liver cancer group were up-regulated by 4.6,7.4 and 10.4-folds,respectively.Immunohistochemistry showed that compared with the control group,the positive rate of Wnt3a in the degeneration group was 66.7%(12/18,χ2=10.701,P=0.001),and both the precancerous and

关 键 词：肝细胞癌 基因表达 肝细胞癌变 WNT3A 早期监测 

分 类 号：R735.7[医药卫生—肿瘤]