脂多糖诱导小鼠肝癌YAP表达及其调控巨噬细胞极化的研究  

Lipopolysaccharide induces YAP expression and regulates macrophage polarization in liver tumor of mouse

作　　者：郑啟发 李华[1] 袁泽南 叶林森[1] 朱曙光[1] 夏龙 Zheng Qifa;Li Hua;Yuan Zenan;Ye Linsen;Zhu Shuguang;Xia Long(Department of Liver Surgery,the Third Affiliated Hospital of Sun Yat-sen University,Guangzhou 510630,China)

机构地区：[1]中山大学附属第三医院肝脏外科,广州510630

出　　处：《中华肝脏外科手术学电子杂志》2020年第3期283-288,共6页Chinese Journal of Hepatic Surgery(Electronic Edition)

基　　金：国家自然科学基金面上项目(81172038);广州市科技计划项目(201607010024)。

摘　　要：目的探讨小鼠肝癌原位移植瘤模型中脂多糖(LPS)诱导Yes相关蛋白(YAP)的表达情况及其调控巨噬细胞极化的机制。方法建立野生型(WT)、YAP基因敲除(YAP-HKO)、YAPWT C57BL/6小鼠肝癌模型,随机选取4只WT小鼠作为LPS组,予每天腹腔注射LPS 5 mg/kg;另选4只作为对照组,予腹腔注射等体积生理盐水。为验证YAP的作用,设置YAP-HKO小鼠组及其同窝YAP-WT小鼠组,每组各4只。各组小鼠饲养10 d后处死,测量肝肿瘤体积;免疫组化法观察LPS组和对照组小鼠肝组织YAP、Ki67、F4/80、CD163,YAP-HKO和YAP-WT小鼠肝组织F4/80、CD163的表达情况。流式细胞术检测小鼠肝组织巨噬细胞分型情况。两组检测指标比较采用t检验。结果LPS组小鼠肝肿瘤体积为(2.50±0.79)cm^3,明显大于对照组的(0.97±0.29)cm^3(t=3.641,P<0.05);YAPHKO小鼠肝肿瘤体积为(0.13±0.11)cm^3,明显小于YAP-WT小鼠的(0.78±0.23)cm^3(t=-5.100,P<0.05)。LPS组肝组织YAP、Ki67、F4/80、CD163表达均高于对照组。YAP-HKO小鼠肝组织F4/80、CD163表达均低于YAP-WT小鼠。LPS组肝组织M2型巨噬细胞百分率为(69±3)%,明显高于对照组的(45±4)%(t=9.768,P<0.05)。YAP-HKO小鼠组肝组织M2型巨噬细胞百分率为(54±4)%,明显低于YAP-WT小鼠组的(68±7)%(t=-3.669,P<0.05)。结论在小鼠肝癌原位移植瘤模型中,LPS通过促进YAP的表达,募集巨噬细胞并促进其向M2型极化,促进肿瘤增殖。Objective To investigate the role of lipopolysaccharide(LPS)in inducing the expression of YAP and regulating the macrophage polarization in the mouse model of orthotopically implanted liver tumor.Methods The wild type(WT),YAP gene-knockout(YAP-HKO),YAP-WT C57BL/6 mouse liver tumor models were established.After the models were established,4 WT mice were randomly selected and assigned into the LPS group and were administered intraperitoneally with LPS 5 mg/kg per day.Another 4 mice were allocated into the control group and were given intraperitoneally with equivalent amount of normal saline.To verify the effect of YAP,YAP-HKO mouse group(n=4)and YAP-WT littermate mouse group(n=4)were established.All mice in each group were sacrificed after feeding for 10 d.The volume of liver tumor was measured.The expression levels of YAP,Ki67,F4/80 and CD163 in liver tissues in LPS and control groups,and the expression of F4/80 and CD163 in liver tissues of YAP-HKO and YAP-WT mice were observed by immunohistochemistry.The phenotype of macrophages in the mouse liver tissues was analyzed by flow cytometry.Parameters between two groups were statistically compared by t test.Results The volume of liver tumor in LPS group was(2.50±0.79)cm^3,significantly larger than(0.97±0.29)cm^3 in control group(t=3.641,P<0.05).The liver tumor volume of YAP-HKO mice was(0.13±0.11)cm^3,significantly smaller than(0.78±0.23)cm^3 of YAP-WT mice(t=-5.100,P<0.05).The expression levels of YAP,Ki67,F4/80 and CD163 in the liver tissues in LPS group were higher than those in control group.The expression levels of F4/80 and CD163 in the liver tissues of YAP-HKO mice were lower compared with those of YAP-WT mice.The percentage of M2-type macrophages in the liver tissues in LPS group was(69±3)%,significantly higher than(45±4)%in the control group(t=9.768,P<0.05).The percentage of M2-type macrophages in the liver tissues of YAP-HKO mice was(54±4)%,significantly lower than(68±7)%of YAP-WT mice(t=-3.669,P<0.05).Conclusions In the mouse model of orthotopicall

关 键 词：肝肿瘤 实验性 脂多糖类 Yes相关蛋白 巨噬细胞 小鼠 

分 类 号：R735.7[医药卫生—肿瘤]